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CHARACTERISTICS OF BUTTER

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 CHARACTERISTICS
OF BUTTER

Butter
Conor Walton

Medical Research in Biblical Times

Examination of Passages from the Bible,
Exactly as Written

Liubov Ben-Nun

NOT FOR SALE

 Nutrition is an important dimension in human health. This study
deals specifically with butter. What is its origin? History? How is
butter made? How is butter preserved? What is its nutritional
value? What are its health effects? What are the adverse effects?
Can butter be contaminated?
Biblical verses dealing with butter were studied from a
contemporary viewpoint.

Author & Editor: Liubov Ben-Nun, Professor Emeritus

Ben Gurion University of the Negev
Faculty of Health Sciences, Dept. of Family Medicine
Beer-Sheva, Israel.

B. N. Publication House. Israel. 2019.

E-Mail: [email protected]

The Author gains no financial or other benefits.

Technical Assistance: Carmela Moshe.

NOT FOR SALE

 CONTENTS
INTRODUCTION 4

BIBLICAL VERSES 6

WHAT IS BUTTER? 7

HISTORY 8

BUTTER VARIETIES 11

MAKING BUTTER 15

HOW TO STORE BUTTER? 26

ALTERNATIVES TO BUTTER 30

NUTRITIONAL FACTS 32
CHANGING COMPOSITION

CONSUMPTION 43

BUTTER ADULTERATION 45

HEALTH EFFECTS 48

CONTAMINATION 88
BACTERIAL AGENTS
VARIOUS POLUTANTS

SUMMARY 107
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INTRODUCTION
James (1) mentioned that nutrition is now becoming once more of
intense interest to biological and medical scientists working on the
control of development and human health. It is also now of ever
greater public health interest. Few scientists, however, recognize
that the same interest for those involved in fundamental science and
public health developed a century ago focusing on the way in which
nutrition and specific micronutrients, as well as general energy and
protein intakes, were crucial to infant growth and appropriate
development. The discovery of vitamins was matched by the
proposition that stunted children in poor communities in the
Western world were suffering from poverty-related poor diets. The
critical role of nutrition was established by feeding studies, which
then led to major food and agricultural policy changes during the
Second World War, when food supplies were scarce throughout
Europe. The success of these wartime policies led to a revolution in
governmental thinking and a cheap food policy, together with a
major boost in national agricultural production as an issue of national
security. Nutritionists transferred their scientific interest to the study
of childhood malnutrition in the developing world. The promotion of
intensive agriculture and the food industry led to a revolution in food
supplies, with the intense promotion of meat, milk, butter, and sugar
production and consumption. The resulting escalation in
cardiovascular disease related to the dietary change slowly altered
public health policies, but as cardiovascular deaths decreased in the
developed world, obesity and diabetes progressively increased. Now
the lower- and middle-income countries (i.e., the developing world)
have far more cardiovascular disease as Western diets and cultural
habits are imported. The remarkable escalation of diabetes and
cardiovascular disease, particularly in populations currently and
previously subjected to malnutrition, now reveals unusual
susceptibility to these diseases. This susceptibility is increasingly
related to the conjunction of fetal malnutrition and later
inappropriate diets. The alarming escalation in the health burden
suggests that two-thirds of the world's population is super-sensitive
to weight gain, diabetes, cardiovascular disease, and perhaps many
cancers. New evidence on epigenetics and the structural changes in
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the fetus in response to inappropriate maternal diets provides

mechanisms to explain this. Unfortunately, a vicious
intergenerational cycle of maternal and fetal epigenetic change
seems to herald markedly increased future burdens of disease. The
nutrition field is therefore challenged not only in terms of science,
but also in new dimensions of public health of immense economic
significance (1).
Cobiac & al. (2) demonstrated the applicability of an original
method to assess the economic and health impacts of compliance
with food-based recommendations. The method takes account of
consumers' preferences and the associated adoption cost in the
assessment of various recommendations. An economic model of diet
choice with an epidemiological model was combined to compute the
health impacts of dietary changes. To demonstrate the use of the
method, the impacts of a 5% variation in the consumption of seven
food groups taken separately was analyzed: a 5% increase in
consumption of fruits and vegetables (F&V) and milk products; and a
5% decrease in consumption of red meat, all meats, salty/sweet
products, ready meals and butter/cream/cheese. A
recommendation, when adopted by consumers, generates important
changes in the whole diet due to substitutions and
complementarities among foods. All simulated recommendations
have a positive impact on health. The F&V recommendation has the
largest impact on the number of DALYs averted, but the highest
adoption cost for consumers, especially for low-income consumers.
Alone, the change in energy intake explains from 71% to 98% of the
DALYs averted induced by a recommendation. The data indicate that
small increases in recommended foods have the potential of
generating relatively significant health gains. Preference-driven
substitutions among foods have a major effect on simulated health
outcomes and should be included in the assessment of dietary
recommendations, together with the adoption cost borne by
consumers (2).

Nutrition is an important dimension in human health. This study deals

specifically with butter. What is its origin? History? How is butter made?
How is butter preserved? What is its nutritional value? What are its
health effects? What are the adverse effects? Can butter be
contaminated?
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Biblical verses dealing with butter were studied from a

contemporary viewpoint.

References
1. James P. Marabou 2005: nutrition and human development. Nutr
Rev. 2006;64(5 Pt 2):S1-11; discussion S72-91.
2. Cobiac L, Irz X, Leroy P, et al. Accounting for consumers' preferences
in the analysis of dietary recommendations. Eur J Clin Nutr. 2018 Sep 25.

BIBLICAL VERSES
"And he took butter, and milk, and the calf which he had dressed, and set
it before them.." (Genesis 18:8).
"He asked water, and she gave him milk; she brought forth butter in a
lordly dish" (Judges 5:25).
"Butter of kine, and milk of sheep, with fat of lambs, and rams of the
breed of Bashan, and goats, with the fat of kidneys of wheat; and thou didst
drink the pure blood of the grape" (Deuteronomy 32:14).
"And honey, and butter, and sheep, and cheese of kine, for David, and for
the people that were with him, to eat: for they said, The people is hungry,
and weary, and thirsty, in the wilderness (2 Samuel 17:29).
"He shall not see the rivers, the floods, the brooks of honey and butter"
(Job 20:17).
"When I washed my steps with butter, and the rock poured me out rivers
of oil" (Job 29:6).
"Surely the churning of milk bringeth forth butter, and the wringing of
the nose bringeth forth blood: so the forcing of wrath bringeth forth strife
(Proverbs 30:33).
"Butter and honey shall he eat, that he may know to refuse the evil, and
choose the good" (Isaiah 7:15).
"And it shall come to pass, for the abundance of milk that they shall give
he shall eat butter: for butter and honey shall every one eat that is left in the
land" (Isaiah 7:22).
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WHAT IS BUTTER
Butter is a product made from the solid components in milk (fat
and protein). Although most often made from cow’s milk, butter can
be made out of milk from sheep, goats, buffalo, or other mammals
(1).
Butter usually consists of approximately 80 percent fat, 15
percent water, and five percent protein. The small amount of protein
in butter acts as an emulsifier allowing the water and fat to stay
suspended in single-phase solution. The unique mixture of fats on
butter allows it to stay solid at room temperature and melt at
approximately 90 degrees Fahrenheit. The natural color of butter
ranges from white to pale yellow depending on the diet of the animal
that produced the milk. Commercial butters are usually colored
yellow with annatto or carotene to fulfill consumers' expectations of
yellow butter (1).
It takes 21 pounds of fresh, wholesome cow’s milk to make each
pound of butter like the pat of butter on waxed paper at a French
farm in Brittany (2).

Butter on Dish Painting

Justin Clayton

Butter is a dairy product. In commercial butter, there is up to 80%

butterfat. This fat is usually solid when the butter is cold. When
warmed, the butterfat turns to liquid. Aside from the butterfat,
there are proteins, water, and in some cases flavors such as salt that
can be found in the butter. The most common source of making
butter is cow’s milk. However, milk from other mammals may also be
used such as sheep, buffalo, goats, and yaks. Ghee is a type of butter
that is purely butterfat and is made through a process called
rendering butter (3).
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References
1. What is butter? Available at thespruceeats.com/what-is-butter-
1328453.
2. Explore the history and making of butter. Available at
webexhibits.org/butter/.
3. Countries Who Consume The Most Butter. Available at
worldatlas.com/articles/countries-who-consume-the-most-butter.html.

HISTORY
Butter – the everlasting delight of the gourmand, the faithful ally
of the culinary arts, the constant symbol of good living. Through time
and across the globe, butter has had a sacred quality. From the
ancient Fertile Crescent to the present day, butter has symbolized
the powerful, life giving and sacred, the good, the happy, the healthy
and pure. It has sustained lives, cultures and civilizations for millennia
(1).
Butter is as old as Western civilization. In ancient Rome, it was
medicinal - swallowed for coughs or spread on aching joints. In India,
Hindus have been offering Lord Krishna tins full of ghee — luscious,
clarified butter — for at least 3,000 years. Butter’s origins are likely
more humble, though. Rumor has it a nomad made the first batch by
accident. He probably tied a sheepskin bag of milk to his horse and,
after a day of jostling, discovered the handy transformation so many
generations have noticed and learned to apply: Churned milk fat
solidifies into something amazing. The oldest known butter-making
technique still in use today is remarkably similar: farmers in Syria skin
a goat, tie the hide up tight, then fill it with milk and begin shaking
(2).
Although some of the earliest records of butter consumption
come from Roman and Arabian sources, Mediterranean people have
always favored oil in their cooking. Butter, it seems, was the fat of
choice for the tribes of northern Europe - so much so that
Anaxandrides, the Greek poet, derisively referred to barbarians from
the north as “butter-eaters.” Climate likely played a key role in
regional tastes, as the cool weather at northern latitudes allowed
people to store butter longer than Mediterranean cultures could. By
the 12th century, the butter business was booming across northern
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Europe. Records show that Scandinavian merchants exported

tremendous amounts each year, making the spread a central part of
their economy. Butter was so essential to life in Norway, for example,
that the King demanded a full bucket every year as a tax (2).
By the middle Ages, eaters across much of Europe were hooked.
Butter was popular among peasants as a cheap source of
nourishment and prized by nobility for the richness it added to
cooked meats and vegetables. For one month out of each year,
however, the mostly Christian Europeans made dough without their
favorite fat. Until the 1600s, butter-eating was banned during Lent.
For northern Europeans without access to cooking oils, meal-making
could be a struggle during the weeks before Easter. Butter proved so
necessary to cooking, in fact, that the wealthy often paid the Church
a hefty tithe for permission to eat the fat during the month of self-
denial. Demand for this perk was so high that in Rouen, in
northwestern France, the Cathedral’s Tour de Beurre — or Butter
Tower — was financed and built with such tithes. Across the English
Channel in Ireland, butter was so critical to the Irish economy that
merchants opened a Butter Exchange in Cork to help regulate the
trade. Today, barrels of ancient Irish butter, which were traditionally
buried in bogs for aging, are among the most common archeological
finds in the Emerald Isle. In France, butter was in such high demand
by the 19th century that Emperor Napoleon III offered a large prize
for anyone who could manufacture a substitute. In 1869, a French
chemist won the award for a new spread made of rendered beef fat
and flavored with milk. He called it “oleomargarine,” later shortened
to just margarine (2).

Butter Made in a Barn

Jan Spanjaert
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Across the Atlantic, butter consumption started with the pilgrims,

who packed several barrels for their journey on the Mayflower.
During the next three centuries, butter became a staple of the
American farm. At the turn of the 20th century, Americans’ annual
consumption was an astonishing 18 pounds of butter per capita —
nearly a stick and a half per person per week! (2).

The Butter Churner

Ivan Gorokhov

The Great Depression and World War II challenged America’s love

affair with butter. The turmoil brought shortages and rationing, and
margarine — now made with vegetable oil and yellow food coloring
— became a cheaper option for American families. Butter
consumption took a nosedive. In addition, dieticians and the USDA
began promoting a low-fat diet in the 1980s, and butter became
déclassé. By 1997, consumption had fallen to 4.1 pounds per capita
per year. Since then, however, butter has staged a comeback.
Researchers have discovered that the ingredients in old-style
margarine are significantly worse for heart health than the saturated
fats found in natural butter. The news has lured more and more
Americans back to their buttery traditions. The passion for
delectable cuisine is bolstering consumption once again as artisanal
butters appear in chilled grocery cases across the country. And at top
restaurants around the globe, chefs are doing extraordinary things
with this millennia-old food, creating an exciting new page in the
history of butter (2).
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Woman Churning Butter

Jean Francois Millet

References
1. Explore the history and making of butter. Available at
webexhibits.org/butter/.
2. History of Butter. Available at butterjournal.com/butter-history.

BUTTER VARIETIES
This chapter deals with various butter types (1-4):

Sweet cream butter is made from cream that has been

pasteurized to kill any bacteria that would normally ferment the
natural sugars in the cream. Sweet cream butter has a light, fresh
flavor and is the most common commercial butter sold in the U.S.

Sweet Cream Butter

Raw Cream Butter has not been pasteurized nor has it been
allowed to ferment. Raw cream butter has a very short shelf life
(approximately 10 days) and is prized for its fresh, clean flavor.
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Raw Milk Butter

Cultured Butter is produced by allowing bacteria to ferment the

sugars in cream prior to churning it into butter. This provides a tart,
tangy, and complex flavor. Cultured butter was the predominant
type of butter prior to refrigeration and pasteurization. Today,
commercial cultured butter is made from cream that has been
pasteurized and then re-inoculated with a specific bacteria strain to
produce fermentation.

Cultured Butter with Raw Milk

Ghee or clarified butter is produced by heating butter until the

water evaporates off and the proteins separate from the fat. The
resulting product is almost 100 percent butterfat. Ghee, which has a
unique flavor, is a popular ingredient in Middle Eastern cuisine.

Ghee Butter

Spreadable Butter. Butter can be quite stiff at refrigerated

temperatures and manufacturers have produced varieties of
spreadable butter to help combat this problem. Spreadable butter is
usually made soft by combining traditional butter with oils, such as
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vegetable oil, that remain liquid at cooler temperatures. Whipping

air or water into butter is another technique used to create a spread
that remains soft at cold temperatures.

Fruit, Vegetable, and Nut Butters. Butter is often used to

describe other spreadable purees that do not contain any dairy
products. Nut butter, such as peanut butter or almond butter, have a
high-fat content and a consistency similar to dairy butter but contain
no milk products. Fruit and vegetable butter, such as apple butter,
are simply pureed fruit or vegetables that have been cooked down to
reduce the moisture content and create a spreadable consistency
closer to that of dairy butter.

Avocado Milkshake Butter

Shea butter. Shea butter is fat that is extracted from the nuts of
the Shea tree. It’s solid at warm temperatures and has an off-white
or ivory color. Shea trees are native to West Africa, and most shea
butter still comes from that region. Shea butter has been used as a
cosmetic ingredient for centuries. Its high concentration of vitamins
and fatty acids — combined with its easy-to-spread consistency —
make it a great product for smoothing, soothing, and conditioning
his/her skin.

Shea Butter
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Mango butter. Mango butter is a soft, creamy butter that has a

very mild scent. It is extracted from the de-shelled fruit kernels of
the mango tree, and then heated and boiled in order to achieve the
ideal consistency. While mango butter shares many of the same uses
as those butters he/she is probably more familiar with, like shea
butter and cocoa butter, its fatty acid profile differs slightly, and it
has a greater level of vitamins and antioxidants. Its rich nutrients
include vitamin A, vitamin C, palmitic acid, arachidic acid, linoleic
acid, oleic acid and stearic acid. That is why it is so effective for
healing and improving the appearance of his/her skin and hair.

Mango Butter

Cocoa butter, also called theobroma oil, pale-yellow, edible

vegetable fat obtained from cocoabeans, having a mild chocolate
flavour and aroma, and is used in the manufacture of chocolate
confections, pharmaceutical ointments, and toiletries. It is valued for
its melting characteristics, remaining brittle at room temperature or
lower but melting just below body temperature. One of the most
stable fats known, cocoa butter contains antioxidants that discourage
rancidity and allow storage life of two to five years. Its use with other
fats improves their stability. Treatment of cocoa butter to remove
aroma and color also removes antioxidants, increasing susceptibility
to rancidity.
Cocoa butter is an important component of eating chocolate.
Although the chocolate liquor used in manufacturing eating
chocolate already contains cocoa butter, an additional amount is
required to produce a molded product that remains firm until
dissolved in the mouth. The amount of fat obtained from hydraulic
pressing of chocolate liquor in cocoa manufacture has become
insufficient for the demands of chocolate production, and whole
cocoa beans or nibs may be processed mainly for their cocoa butter
content. Because of the high cost of cocoa butter, substitutes have
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been developed, including a type suitable for warm weather because

of its higher melting point; such fats, which many countries prohibit
in products sold as pure chocolate, impart waxy texture and taste
similar to that of cocoa butter.

Raw Cocoa Butter

References
1. What is butter? Available at thespruceeats.com/what-is-butter-
1328453.
2. Kathryn Watson, Kristeen Cherney. Available at
healthline.com/health/beauty-skin-care/what-is-shea-butter.
3. Susan Patterson. 9 Reasons You Should Start Putting Mango Butter
On Your Skin & Hair. Available at naturallivingideas.com/mango-butter/.
4. Cocoa butter. Encyclopædia Britannica. Available at
britannica.com/topic/cocoa-butter.

MAKING BUTTER
First, the cream is separated from fresh whole milk. The cream is
then churned by shaking or beating it vigorously until it thickens. The
remaining liquid (buttermilk) is removed. The clustered butter is
washed and formed into its solid shape and presto! Fresh milk from
dairy farms is collected and brought to the creamery. The cream is
then separated from the fresh whole milk using centrifugal force. It
is then pasteurized by heating it rapidly to a high temperature to
eliminate potential disease-causing bacteria and help the butter stay
fresh longer (1).
Once pasteurized, the cream is beaten vigorously in a churning
cylinder until it thickens naturally into butter. The remaining liquid
(buttermilk) is drained off, and the butter is mixed and blended. At
this point, salt is sometimes added (1).
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Butter from Raw Milk

The final product is, by regulation, at least 80% fat, about 16%
water and 3% milk solids. After being weighed, cut, wrapped and
chilled, the butter is delivered to his/her grocery store, ready for
his/her to add to his/her favorite foods (1).
Megnanou & Niamke (2) mentioned that industrials interest in
fats as raw material, resides in their exceptional quality and
potentialities of exploitation in several fields. This study aimed to
exalt the optimized shea butter quality and present its wide
potentialities of utilization. Hence, the characteristics of beige and
yellow optimized shea butters were determined. Both samples
recorded very weak acid (0.280 ± 0.001 and 0.140 ± 0.001 mgKOH/g)
and peroxide (0.960 ± 0.001 and 1.010 ± 0.001 mEgO2/kg) indexes,
when the iodine indexes (52.64 ± 0.20 and 53.06 ± 0.20 gI2/100 g)
and the unsaponifiable matters (17.61 ± 0.01 and 17.27 ± 0.01%)
were considerable. The refractive indexes (1.454 ± 0.00 and 1.453 ±
0.00) and the pH (6.50 ± 0.30 and 6.78 ± 0.30) were statistically
similar; but the specific gravity (0.915 ± 0.01-0.79 ± 0.01 and 0.94 ±
0.01-0.83 ± 0.01) and the viscosity (90.41 ± 0.20-20.02 ± 0.20 and
125.37 ± 0.20-23.55 ± 0.20 MPas) differed and decreased
exponentially with the temperature increasing (35-65°C), except for
the specific gravity of the yellow butter which decreased linearly.
The UV-Vis spectrum showed a high peak at 300 nm and a rapid
decrease from 300 to 500 nm when the near infra-red one, revealed
peaks at 450, 1200, 1400, 1725 and 2150 nm for all the samples. The
chromatographic profile identified palmitic (16.42 and 26.36%),
stearic (32.39 and 36.36%), oleic (38.12 and 29.09%), linoleic (9.72
and 5.92%) and arachidic (1.84 and 1.59%) acids, and also exaltolide
compound (1.51 and 0.68%). The samples also contained essential
minerals (calcium, magnesium, zinc, iron, etc.) carotene (550 ± 50
and 544 ± 50 ppm), vitamins A (0.065 ± 0.001 and 0.032 ± 0.001
µg/g) and E (2992.09 ± 1.90 and 3788.44 ± 1.90 ppm) in relatively
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important amounts; neither microbiological germs nor heavy were

detected. All these valorizing characteristics would confer to the
optimized shea butters good aptitude for exportation and
exploitation in food, cosmetic and pharmaceutical industries (2).
Skelhon & al. (3) noticed that reducing the fat content of
chocolate formulations is a major challenge for the confectionery
industry. The suspension of aqueous microgel agar particles of up to
80% v/v within sunflower oil, cocoa butter, and ultimately chocolate
is reported. The optimized emulsification process involves a shear-
cooling step. The versatility of the method was demonstrated when
applied to white, milk, and dark chocolate formulations, whilst
preserving the desired polymorph V of the cocoa butter matrix. In
addition, this technology can be used as a strategy to disperse
alcoholic beverages into chocolate confectionery (3).
Tondhoosh & al. (4) manipulated individual factors, which
interfere in the continuous churning, to enhance the rheological
properties and chemical composition of butter. This process leads to
achieve softer; more spreadable, and ultimately healthier product for
consumers. In addition it could prevent hardening of texture
especially in winter. Firstly, pasteurized cream with different fat
contents (40 & 45% fat) was passed through heat treatments, and
then it was injected to a continuous churn. Textural and melting
behavior and fatty acid composition of butter were analyzed.
Increasing the fat content of cream (from 40 to 45%) and holding
time (from 3 hours to 5 hours) in mid-temperature (18 C) and
reducing the churning temperature (from 12 C to 10 C), resulted in
soft butter texture and improved butter spreadability. Loss Tangent
(tan δ) was increased from 0.11 to 0.74 (T=15 C; f=1Hz). The melting
temperature of butter was decreased from 36 C to 32 C and total
trans fatty acid content was decreased from 3.2% to 1.87%. It was
concluded that such heating process (which has been studied and
reported in patents) absorbs the low- solid fat content (SFC) fats of
the cream, integrates them into the butter texture a softer and more
spreadable product (4).
Wei & al. (5) reported that Cocoa butter (CB) extracted from
cocoa beans (Theobroma cacao) is the main raw material for
chocolate production, but CB supply is insufficient due to the
increased chocolate demand and limited CB production. CB is mainly
composed of three different kinds of Triacylglycerols (TAGs), 1,3-
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dipalmitoyl-2-oleoyl-glycerol (POP, C16:0-C18:1-C16:0), 1-palmitoyl-

3-stearoyl-2-oleoyl-glycerol (POS, C16:0-C18:1-C18:0) and 1,3-
distearoyl-2-oleoyl-glycerol (SOS, C18:0-C18:1-C18:0). In general,
Saccharomyces cerevisiae produces TAGs as storage lipids, which
consist of C16 and C18 fatty acids. However, cocoa butter-like lipids
(CBL), which are composed of POP, POS and SOS) are not among the
major TAG forms in yeast. TAG biosynthesis is mainly catalyzed by
three enzymes: Glycerol-3-phosphate Acyltransferase (GPAT),
lysophospholipid acyltransferase (LPAT) and diacylglycerol
acyltransferase (DGAT), and it is essential to modulate the yeast TAG
biosynthetic pathway for higher CBL production. Seven GPAT genes
and three LPAT genes were cloned from cocoa cDNA, in order to
screen for CBL biosynthetic gene candidates. By expressing these
cloned cocoa genes and two synthesized cocoa DGAT genes in S.
cerevisiae, total fatty acid production, TAG production and CBL
production in some of the strains were increased. In the best
producer, the potential CBL content was eightfold higher than the
control strain, suggesting the cocoa genes expressed in this strain
were functional and might be responsible for CBL biosynthesis.
Moreover, the potential CBL content increased 134-fold over the
control Y29-TcD1 (IMX581 sct1Δ ale1Δ lro1Δ dga1Δ with TcDGAT1
expression) in strain Y29-441 (IMX581 sct1Δ ale1Δ lro1Δ dga1Δ with
TcGPAT4, TcLPAT4 and TcDGAT1 expression) further suggesting
cocoa GPAT and LPAT genes functioned in yeast. The data show that
cocoa TAG biosynthetic genes functioned in S. cerevisiae and
identified cocoa genes that may be involved in CBL production.
Moreover, expression of some cocoa CBL biosynthetic genes
improved potential CBL production in S. cerevisiae, showing that
metabolic engineering of yeast for cocoa butter production can be
realized by manipulating the key enzymes GPAT, LPAT and DGAT in
the TAG biosynthetic pathway (5).
Liang & Were (6) revealed that sunflower butter use as an
allergen-free alternative to tree and legume nut butter in baking is
limited by chlorogenic acid induced greening that occurs at alkaline
pH. Limited information is available on controlling this greening in a
food matrix. This study examined how different liquid sweeteners
and relative humidity influenced greening of sunflower butter
cookies. Doughs had similar initial pH (7.52-7.66) which increased to
8.44-9.13 after baking as ranked: xylitol>maple syrup>corn
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syrup>honey>agave syrup. Cookies made with maple syrup had the

highest moisture and greening corresponding with lowest free
chlorogenic acid. The % greening followed the same trend as
greening intensity, and was positively correlated (r=0.9101) with
chlorogenic-lysine adduct content. The findings provide an
ingredient solution to controlling greening, as results demonstrate
that greening can be promoted with high relative humidity storage,
and use of high moisture and pH ingredients. Unwanted greening
can be inhibited by simply changing the liquid sweetener (6).
Rogers & al. (7) stated that Chlorogenic acid (CGA) binding to
proteins in alkaline conditions results in the production of green
trihydroxy benzacradine (TBA) derivatives. The formation of TBA
derivatives could decrease product quality due to the potential losses
in soluble protein and antioxidants and the production of an
undesirable green color. To determine how cookie formulation
affected the formation of TBA derivatives in sunflower butter
cookies, two egg replacers (chia and banana) and two baking
temperatures (162.8 and 190.6 C) were used. Moisture, greening
intensity, CGA content and antioxidant capacity were measured.
Cookies made with egg and baked at 162.8°C had the highest
moisture, internal greening intensity, and TBA derivative formation,
in addition to lower CGA content and antioxidant capacity. Cookies
made with banana baked at 190.6°C produced the opposite outcome
with 35, 4, and 23% less internal greening, moisture, and TBA
derivatives, respectively, and 90 and 76% higher CGA and antioxidant
capacity. Internal greening was positively correlated with moisture
and adduct concentration, and negatively correlated with spread
factor and CGA content. Moisture had a significant impact on
greening, which indicates that baking temperature and cookie dough
formulation can be modified to produce a less green cookie with
more unreacted antioxidants and protein (7).
Gorrepati & al. (8) mentioned that during the last few years the
popularity for the plant based butters (nut and seed butters) has
increased considerably. Earlier peanut butter was the only
alternative to the dairy butter, but over the years development in the
technologies and also the consumer awareness about the plant
based butters, has led the development of myriad varieties of butters
with different nuts and seeds, which are very good source of protein,
fiber, essential fatty acids and other nutrients. These days' different
 11

Ben-Nun L butter

varieties of plant based butters are available in the market viz.,

peanut butter, soy butter, almond butter, pistachio butter, cashew
butter and sesame butter etc. The form of butter is one of the
healthy way of integrating nuts and seeds in to our regular diet. Nut
and seed butters are generally prepared by roasting, grinding and
refrigerated to consume it when it is still fresh. During this process it
is imperative to retain the nutritional properties of these nuts and
seeds in order to reap the benefits of the fresh nuts and seeds in the
form of butter as well. Proper care is needed to minimize the
conversion of healthful components in to unhealthy components
during processing and further storage. Roasting temperature,
temperatures during grinding and storage are the vital factors to be
considered in order to have healthy and nutritious plant based
butters. In this article, different plant based butters and their
processing methods have been described (8).

Spiridonov & al. (9) discussed the results of study on

substantiating the possibilities of the use of flavolignans of Silybum
marianum (L.) Gaertn with a purpose to create a new fat product -
butter fortified by an "Liguid extract Silybum marianum (L.) Gaertn."
are being discussed. Slowdown of peroxidation process is revealed in
fortified butter in comparison with ordinary butter. With the help of
spectrometry method higher content of flavolignans was detected in
fortified butter (converting to silibin - the main component of "Liguid
Exstract Silybum marianum (L) Gaertn"), what allows to speak about
increase of its potential therapeutic effect (9).
Shekarchizadeh & al. (10) prepared Cocoa butter analog from
camel hump fat and tristearin by enzymatic interesterification in
supercritical carbon dioxide (SC-CO2) using immobilized
Thermomyces lanuginosus lipase (Lipozyme TL IM) as a biocatalyst.
 10

Ben-Nun L butter

Optimal process conditions were determined using neural networks

and genetic algorithm optimization. Response surfaces methodology
was used to design the experiments to collect data for the neural
network modeling. A general regression neural network model was
developed to predict the response of Triacylglycerol (TAG)
distribution of cocoa butter analog from the process pressure,
temperature, tristearin/camel hump fat ratio, water content, and
incubation time. A genetic algorithm was used to search for a
combination of the process variables for production of most similar
cocoa butter analog to the corresponding cocoa butter. The
combinations of the process variables during genetic algorithm
optimization were evaluated using the neural network model. The
pressure of 10 MPa; temperature of 40°C; SSS/CHF ratio of 0.6:1;
water content of 13% (w/w); and incubation time of 4.5 hours was
found to be the optimum conditions to achieve the most similar
cocoa butter analog to the corresponding cocoa butter (10).
Lamothe & al. (11) applied a washing treatment to caprine cream
before churning in order to improve phospholipids and MFGM
protein purification from buttermilk and butter serum. Cream
obtained from a first separation was diluted with water and
separated a second time using pilot plant equipment. Regular and
washed creams were churned to produce buttermilk and butter,
from which butter serum was extracted. The washing treatment
allowed a significant decrease of the casein content. As a result, the
phospholipids-to-protein ratios in washed buttermilk and butter
serum were markedly increased by 2.1 and 1.7-folds respectively,
which represents an advantage for the production of phospholipids
concentrates. However, when compared with bovine cream, lower
phospholipids-to-protein ratios were observed when the washing
treatment was applied to caprine cream. A higher concentration of
MFGM protein and a lower retention of phospholipids during
washing treatment are responsible for the lower phospholipids-to-
protein ratios in buttermilk and butter serum obtained from caprine
cream. The phospholipids distribution in the butter making process
was similar to the one obtained from bovine regular and washed
cream. Phospholipids were preferentially concentrated in the butter
serum rather than the buttermilk fraction. This simple approach
permitted the production of caprine and bovine butter sera extracts
 11

Ben-Nun L butter

containing up to 180 and 240 g phospholipids/kg sera, respectively,

on a dry basis (11).
Sagiri & al. (12) mentioned that the search for cocoa butter
equivalents in food and pharmaceutical industries has been gaining
importance. In the present study, mango butter was explored as
cocoa butter equivalent. Aqueous gelatin solution (20% w/w)
containing cocoa butter and mango butter water-in-oil (fat) type
emulsion gels were prepared by hot emulsification method. XRD and
DSC melting profiles suggested the presence of unstable polymorphic
forms (α and β') of fats in the emulsion gels. The crystal size and
solid fat content analyses suggested that the presence of aqueous
phase might have hindered the transformation of unstable
polymorphic forms to stable polymorphic form (β) in the emulsion
gels. Fat crystals in the emulsion gels were formed by instantaneous
nucleation via either uni- or bidimensional growth (Avrami analysis).
The viscoelastic nature of the emulsion gels was evaluated by
modified Peleg's analysis (stress relaxation study). Results inferred
that the physical, thermal, and mechanical properties of mango
butter emulsion gels are comparable to those of cocoa butter
emulsion gels. On the basis of preliminary studies, it was suggested
that the mango butter emulsion gels may have potential to be used
as cocoa butter equivalents (12).
Wagener & Kerr (13) noticed that it has been difficult to produce
acceptable pecan butters as the high oil content results in a product
that flows and separates too easily. The objective of this work was to
create pecan butters with varying oil levels (50-70%) and determine
which would give the most acceptable product. Consumers rated
pecan butters with 55-60% oil the most acceptable, whether roasted
or not. Acceptability varied most in terms of texture and
spreadability, but not flavor. Under large deformation firmness
varied from 51.8 g (70% oil) to 4,880 g (50%) oil, while "spreadability"
ranged from 19.2 to 7748 (g/s). Samples with 70% oil had the lowest
viscosity and were Newtonian. Pecan butters with 50-55% oil had
high viscosity and were shear thinning. Yield stress decreased with
oil content, ranging from 0.014 to 500 Pa. The storage modulus (G')
increased from ∼7 Pa for samples with 70% oil up to 260,000 Pa for
those with 50% oil. In conjunction, tan δ decreased from 1 to 0.07,
showing the products take on much more solid-like behavior as oil is
removed. In conclusion, the rheological properties of pecan butter
 12

Ben-Nun L butter

were quite sensitive to the amount of oil in the product. Differences

in acceptability were primarily due to "texture" and "spreadability,"
suggesting there is a limited range of firmness and spreadability that
consumers will deem acceptable. There has been considerable
demand for butters and spreads made from a variety of culinary nuts.
Pecans generally have too much oil (∼70%) to make a product with
proper consistency and stability. In this study, some of the oil was
removed to overcome this problem. It was found that pecan butter
with 55-60% oil was most acceptable to consumers and with the level
of firmness, yield stress, and spreadability most similar to commercial
nut butters. The oil was relatively simple to remove from unroasted
nuts, thus manufacturers could easily produce more acceptable
pecan butter for the market (13).
Brents & al. (14) presented the data on the chemical composition
and the technology of manufacturing a new sort of butter for child's
and dietetic nutrition. The butter has high biological value due to the
introduction of Polyunsaturated Fatty Acids (PUSFA) with vegetable
oils and milk-protein ingredients. The milk-protein ingredients also
play the role of the butter structure stabilizers. The chemical
composition of the new sort of butter, including the vitamin and
mineral content, as well as the amino-acid composition of the butter
plasma are described. It is shown that the content of PUSFA in the
new sort of butter is 10-fold higher than in the routine butter.
Various kinds of dessert butter have been developed with different
flavoring additives. One of the butter variant has been enriched with
bifidobacteria. The new sort of butter is recommended for the
dietetic nutrition of children and adults suffering from obesity, as
well as for the nutrition of the middle- and old-aged subjects to
prevent lipid metabolism disorders (14).
Jahurul & al. (15) investigated the blending effects of mango seed
fat (MSF), extracted using supercritical fluid, and palm stearin (PS) to
formulate hard cocoa butter replacers (CBRs). The triglycerides (TG),
thermal properties and solid fat content (SFC) of the formulated
blends were determined using different chromatographic and
thermal techniques. All the blends had three main TGs; namely, 1,3-
dipalmitoyl-2-oleoylglycerol (POP) (8.6-17.7%), 1-palmitoyl-2-oleoyl-
3-stearoyl-glycerol (POS) (12.6-19.6%), and 1,3-distearoyl-2-oleoyl-
glycerol (SOS) (37.2-31.4%), with SOS being the major component.
The melting peak temperatures gradually increased and shifted
 13

Ben-Nun L butter

towards higher temperatures with PS. The crystallization onset

temperatures increased, while the offset decreased with PS. The SFC
did not drop to 0% at 37.5°C, which was shifted to 0% at and above
40°C for some blends. The studies revealed that CBRs could be
prepared by blending MSF and PS, and they could be utilized by
chocolate manufacturers in tropical countries (15).
Ma & al. (16) noticed that Cinnamomum camphora trees have a
vast range of distribution in southern China and the seed oil has
unique fatty acid (FA) properties and various bio-activities. In this
work, Cinnamomum camphora Seed Oil (CCSO) was utilized to
synthesize value-added cocoa butter substitute (CBS) by enzymatic
interesterification. The synthesis was conducted in a solvent-free
system by blending CCSO with fully hydrogenated palm oil under the
catalysis of Lipozyme RM IM. The reacted products were assessed
with physicochemical properties, i.e. FA composition, Slip Melting
Point (SMP), triacylglycerol (TAG), crystal polymorphism,
microstructure, melting and crystallization properties and solid fat
content (SFC). It showed that MCFAs (capric acid plus lauric acid) was
the main fatty acid in products, accounting for over 45%. Comparing
to physical blends, some novel TAG species such as LaLaLa and
LaMLa/LaLaM were observed after enzymatic interesterification
whereas SSS TAGs were reduced. IP presented a ball-like, well-
distributed and nearly round crystal microstructure and a smaller
crystal size. Moreover, it should be mentioned that SFC of IP ranging
from 31.85 to 38.47% at 25°C with most β' crystal forms, was
beneficial to improve the spreadability in term of confectionery
products and baked goods. The SMP of the interesterified products
was 35.75-36.15 C which closed to the commercial CBS. Hence, the
products synthesized can be used to as CBS, and the results in this
study also showed CCSO have value-added applications (16).

This chapter (1-16) presents various processing methods of

various types of butters.

References
1. How Butter is Made. Available at dairygoodness.ca/butter/how-
butter-is-made.
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2. Megnanou RM, Niamke S. Improving the optimized shea butter

quality: a great potential of utilization for common consumers and
industrials. Springerplus. 2015 Nov 4;4:667.
3. Skelhon TS, Olsson PK, Morgan AR, Bon SA. High internal phase agar
hydrogel dispersions in cocoa butter and chocolate as a route towards
reducing fat content. Food Funct. 2013;4(9):1314-21.
4. Tondhoosh A, Nayebzadeh K, Mohamadifar MA, et al. Industrial
application of different heat treatments and cream fat contents for
improving the spreadability of butter. Recent Pat Food Nutr Agric. 2016;
8(2):107-15.
5. Wei Y, Bergenholm D, Gossing M, et al. Expression of cocoa genes in
Saccharomyces cerevisiae improves cocoa butter production. Microb Cell
Fact. 2018;17(1):11.
6. Liang S, Were LM. Chlorogenic acid oxidation-induced greening of
sunflower butter cookies as a function of different sweeteners and storage
conditions. Food Chem. 2018;241:135-42.
7. Rogers A, Hahn L, Pham V, Were L. Greening in sunflower butter
cookies as a function of egg replacers and baking temperature. J Food Sci
Technol. 2018;55(4):1478-88.
8. Gorrepati K, Balasubramanian S, Chandra P. Plant based butters. J
Food Sci Technol. 2015;52(7):3965-76.
9. Spiridonov AM, Popov AD, Kurkin VA, et al. The use of flavolignans
of "Silybum marianum (L.) Gaertn." to create a new fat product – fortified
butter. Vopr Pitan. 2005;74(6):42-5.
10. Shekarchizadeh H, Tikani R, Kadivar M. Optimization of cocoa
butter analog synthesis variables using neural networks and genetic
algorithm. J Food Sci Technol. 2014;51(9):2099-105.
11. Lamothe S, Robitaille G, St-Gelais D, Britten M. Butter making from
caprine creams: effect of washing treatment on phospholipids and milk fat
globule membrane proteins distribution. J Dairy Res. 2008;75(4):439-43.
12. Sagiri SS, Sharma V, Basak P, Pal K. Mango butter emulsion gels as
cocoa butter equivalents: physical, thermal, and mechanical analyses. J
Agric Food Chem. 2014;62(47):11357-68.
13. Wagener EA, Kerr WL. Effects of oil content on the sensory,
textural, and physical properties of pecan butter (Carya illinoinensis). J
Texture Stud. 2018;49(3):286-92.
14. Brents MIa, Fursova SA, Baeva VS, et al. A new type of creamery
butter for pediatric and dietetic nutrition. Vopr Pitan. 1987;(3):55-8.
15. Jahurul MH, Zaidul IS, Nik Norulaini NA, et al. Hard cocoa butter
replacers from mango seed fat and palm stearin. Food Chem. 2014;
154:323-9.
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16. Ma X, Hu Z, Mao J, et al. Synthesis of cocoa butter substitutes from

Cinnamomum camphora seed oil and fully hydrogenated palm oil by
enzymatic interesterification. J Food Sci Technol. 2019; 56(2):835-45.

HOW TO STORE BUTTER?

Keep butter refrigerated in its original wrapper. The foil
laminated paper helps prevent spoilage from exposure to light and
air, and also protects butter from picking up the flavor of other foods.
If he/she wraps it well, once opened, both salted and unsalted butter
will keep in the fridge for three weeks. However, if he/she plans to
keep it longer than a few weeks, butter will keep its fresh taste better
if you wrap it again in extra foil or plastic. Butter freezes well, but
should be further protected by over-wrapping it in additional foil or
heavy-duty plastic wrap or a freezer bag. Properly wrapped, salted
butter will keep in the freezer for up to one year. After this, it may
begin to lose the fresh butter flavor and pick up other odors or
flavors from the freezer. Unsalted butter will keep in the freezer for
up to three months (1).
At refrigerated temperatures, butter will stay fresh for up to four
months. Butter can also be frozen and kept fresh for up to a year. At
room temperature, the length of freshness will depend on the
butter’s exposure to light and oxygen, but butter will generally stay
fresh for several days unrefrigerated. Covering butter in a ceramic
dish or butter bell will help preserve freshness at room temperature
by reducing oxygen exposure and heat. Butter that has a strong
smell or a bitter or sharp flavor has most likely gone rancid and
should be discarded (1).

Krause & al. (2) mentioned that butter is often stored for
extended periods of time; therefore, it is important for
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Ben-Nun L butter

manufacturers to know the refrigerated and frozen shelf life. The

objectives of this study were to characterize the effect of refrigerated
and frozen storage on the sensory and physical characteristics of
butter. Fresh butter was obtained on 2 occasions from 2 facilities in
113-g sticks and 4-kg bulk blocks (2 facilities, 2 package forms).
Butters were placed into both frozen (-20 degrees C) and refrigerated
storage (5 degrees C). Frozen butters were sampled after 0, 6, 12, 15,
and 24 months; refrigerated butters were sampled after 0, 3, 6, 9, 12,
15, and 18 months. Every 3 months, oxidative stability index (OSI)
and descriptive sensory analysis (texture, flavor, and color) were
conducted. Every 6 months, peroxide value (PV), free fatty acid value
(FFV), fatty acid profiling, vane, instrumental color, and oil turbidity
were examined. A mixed-model ANOVA was conducted to
characterize the effects of storage time, temperature, and package
type. Storage time, temperature, and package type affected butter
flavor, OSI, PV, and FFV. Refrigerated butter quarters exhibited
refrigerator/stale off-flavors concurrent with increased levels of
oxidation (lower oxidative stability and higher PV and FFV) within 6
months of refrigerated storage, and similar trends were observed for
refrigerated bulk butter after 9 months. Off-flavors were not evident
in frozen butters until 12 or 18 months for quarters and bulk butters,
respectively. Off-flavors in frozen butters were not correlated with
instrumental oxidation measurements. Because butter is such a
desirable fat source in terms of flavor and textural properties, it is
important that manufacturers understand how long their product can
be stored before negative attributes develop (2).
Lozano & al. (3) analyzed the major aroma compounds of
commercial sweet cream AA butter quarters were analyzed by GC-
olfactometry and GC-MS combined with Dynamic headspace analysis
(DHA) and Solvent-assisted flavor evaporation (SAFE). In addition,
the effect of long-term storage (0, 6, and 12 months) and type of
wrapping material (wax parchment paper vs. foil) on the aroma
components and sensory properties of these butters kept under
refrigerated (4 degrees C) and frozen (-20 degrees C) storage was
evaluated. The most intense compounds in the aroma of pasteurized
AA butter were butanoic acid, delta-octalactone, delta-decalactone,
1-octen-3-one, 2-acetyl-1-pyrroline, dimethyl trisulfide, and diacetyl.
The intensities of lipid oxidation volatiles and methyl ketones
increased as a function of storage time. Refrigerated storage caused
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Ben-Nun L butter

greater flavor deterioration compared with frozen storage. The

intensity and relative abundance of styrene increased as a function of
time of storage at refrigeration temperature. Butter kept frozen for
12 months exhibited lower styrene levels and a flavor profile more
similar to that of fresh butter compared to butter refrigerated for 12
months. Foil wrapping material performed better than wax
parchment paper in preventing styrene migration into butter and in
minimizing the formation of lipid oxidation and hydroxyl acid
products that contribute to the loss of fresh butter flavor (3).
Mohd & al. (4) evaluated the storage stability of preservative-free
peanut butter for changes in physicochemical quality including
moisture content and water activity, microbiological properties,
oxidative stability and textural quality in terms of spreadability and
firmness. The study was conducted for 16 weeks at storage
temperature of 10, 25 and 35 C on natural and pure peanut butter
produced from two varieties of peanuts, the Virginia and Spanish
TMV-2 varieties of China and India origin, respectively. The peanuts
were ground using a high speed grinder for 2.5 and 3.0 min to
produce peanut butter without addition of other ingredient. The
natural peanut butter exhibited stability and had acceptable
microbial count during storage. Storage at 10°C gave similar textural
quality with commercial product until week 8 and without
appreciable loss in oxidative stability until week 12. At higher storage
temperatures of 25 and 35 C, oxidative stability was shortened to 4
weeks of storage. Among the factors of storage temperature and
time, grinding time and peanut variety, storage temperature had the
most significant effects on quality changes of natural peanut butter
(4).
Atonfack & al. (5) stated that sunflower seed derived butter can
be a source of protein and phenolic antioxidants in refrigerated
dough. Chlorogenic quinone-amino acid induced greening can
however occur at alkaline pH, which could result in less bioavailable
conjugated phenol-amino acids. Acidulants were tested as potential
anti-greening ingredients in refrigerated chemically leavened cookie
dough. Effect of refrigerated storage time, leavening agents and
acidulants on tryptophan fluorescence (λex = 280 nm, λem = 300-500
nm), color (hunter L*, a*, b* color scale), reducing capacity [1,1'-
diphenyl-2-picryl-hydrazyl (DPPH) and Folin-Ciocalteu reducing
capacity (FCRC)], and hydroxycinnamic acids were measured. The pH
 18

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range of acidified doughs was 4.83-6.98 compared to 7.65-9.18 in

non-acidified leavened doughs after 24 days. Greening was higher in
baking soda dough control (a*=-0.54) than baking powder dough
control (a*= 2.98) after 24 days, attributed to higher pH (9.18) of the
former compared to pH 7.14 in the later. Tryptophan fluorescence
intensity in baking soda dough decreased in the order: control >
glucono-delta lactone ≈ citric acid after 24 days. The DPPH and FCRC
of acidified doughs were greater than corresponding control doughs.
The data show that the use of acidulants would prevent greening in
sunflower dough without lowering its phenolic concentration, making
use of sunflower butter in refrigerated dough for baked goods
feasible (5).
Honfo & al. (6) mentioned that storage conditions are key
constraints for quality assurance of the shea (Vitellaria paradoxa
Gaertner) butter. In the Sudan savannah Africa, storage conditions of
butter produced by women vary across and among processors,
traders and consumers. These conditions could impact the quality of
the products and reduced their access to international market. The
present study attempted to investigate the effect of storage duration
and packaging materials on microbiological and physicochemical
characteristics of shea butter under tropical climatic conditions. Five
packaging materials traditionally used in shea butter value chain
were tested for their efficacy in storing shea butter freshly produced.
Total germs, yeasts and mould varied with packaging materials and
storage duration. After 2 months of storage, moisture content of
butter remained constant (5%) whereas acid value increased from 3.3
to 5.4 mg KOH/g, peroxide value from 8.1 to 10.1 meq O2/kg and
iodine value dropped from 48.8 to 46.2 mg I2/100 g in shea butter
irrespectively to the storage materials used. The basket papered
with jute bag was the less effective in ensuring the quality of butter
during storage while plastic containers and plastic bags seemed to be
the best packaging materials (6).

This chapter (1-6) deals with different storage conditions of

different types of butter.
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References
1. How to store butter. Available at dairygoodness.ca/butter/how-to-
store-butter.
2. Krause AJ, Miracle RE, Sanders TH, et al. The effect of refrigerated
and frozen storage on butter flavor and texture. J Dairy Sci. 2008;
91(2):455-65.
3. Lozano PR, Miracle ER, Krause AJ, et al. Effect of cold storage and
packaging material on the major aroma components of sweet cream
butter. J Agric Food Chem. 2007;55(19):7840-6.
4. Mohd Rozalli NH, Chin NL, Yusof YA, Mahyudin N. Quality changes
of stabilizer-free natural peanut butter during storage. J Food Sci Technol.
2016;53(1):694-702.
5. Atonfack JT, Ataman ZA, Were LM . Acidulant effect on greening,
reducing capacity, and tryptophan fluorescence of sunflower buttercookie
dough during refrigerated storage. J Sci Food Agric. 2019;99(5):2186-93.
6. Honfo F, Hell K, Akissoé N, et al. Effect of storage conditions on
microbiological and physicochemical quality of shea butter. J Food Sci
Technol. 2011;48(3):274-9.

ALTERNATIVES TO BUTTER
MANGIFERA SYLVATICA
Akhter & al. (1) stated that Cocoa butter is the pure butter
extracted from cocoa beans and is a major ingredient in the
chocolate industry. Global production of cocoa is in decline due to
crop failure, diseases and ageing plantations, leading to price
fluctuations and the necessity for the industry to find high quality
cocoa butter alternatives. This study explored the potential of a wild
mango (Mangifera sylvatica), an underutilized fruit in south-east Asia,
as a new Cocoa Butter Alternative (CBA). Analyses showed that wild
mango butter has a light colored fat with a similar fatty acid profile
(palmitic, stearic and oleic acid) and triglyceride profile (POP, SOS and
POS) to cocoa butter. Thermal and physical properties are also
similar to cocoa butter. Additionally, wild mango butter comprises
65% SOS (1, 3-distearoyl-2-oleoyl-glycerol) which indicates potential
to become a Cocoa Butter Improver (an enhancement of CBA). It is
concluded that these attractive properties of wild mango could be
prompted by a coalition of policy makers, foresters, food industries
 20

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and horticulturists to promote more widespread cultivation of this

wild fruit species to realize the market opportunity (1).

Reference
1. Akhter S, McDonald MA, Marriott R. Mangifera sylvatica (Wild
Mango): a new cocoa butter alternative. Sci Rep. 2016 Aug 24;6:32050.

MARGARINE
Butter is a dairy product, made from milk or cream. When the
liquid is churned, the butterfat separates from the buttermilk,
making a solid yet spreadable light yellow substance. But butter is
not purely fat; it is about 20 percent water and also contains milk
proteins (solids). “Light” butter contains more water, less fat and
calories (1).
Margarine is an oil-based product. It is typically made of 80
percent vegetable oil and water and flavored to taste like butter.
(Most margarines contain trace amounts of dairy. If he/she is looking
for a completely non-dairy product, choose “vegan margarine”).
He/she can buy margarine in sticks or tubs. Butter-flavored spreads
are sold in tubs and usually contain less oil and more water, for a
softer consistency (1).

Margarine

Prättälä & al. (2) mentioned that Finland is known for a sharp
decrease in the intake of saturated fat and cardiovascular mortality.
Since 2000, however, the consumption of butter-containing spreads -
an important source of saturated fats - has increased. This study
examined social and health-related predictors of the increase among
Finnish men and women. A representative random sample of adult
Finns, invited to a health survey in 2000, was followed up for 11
 21

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years. Altogether 5,414 persons aged 30-64 years at baseline in 2000

were re-invited in 2011. Of men 1,529 (59 %) and of women 1853
(66 %) answered the questions on bread spreads at both time points.
Respondents reported the use of bread spreads by choosing one of
the following alternatives: no fat, soft margarine, butter-vegetable oil
mixture and butter, which were later categorized into margarine/no
spread and butter/butter-vegetable oil mixture (= butter). The
predictors included gender, age, marital status, education,
employment status, place of residence, health behaviors, BMI and
health. Multinomial regression models were fitted. Of the 2,582
baseline margarine/no spread users, 24.6% shifted to butter. Only a
few of the baseline sociodemographic or health-related determinants
predicted the change. Finnish women were more likely to change to
butter than men. Living with a spouse predicted the change among
men. The data show that the change from margarine to butter
between 2000 and 2011 seemed not to be a matter of compliance
with official nutrition recommendations. Further longitudinal studies
on social, behavioral and motivational predictors of dietary changes
are needed (2).

References
1. Lisa Cericola. What’s The Difference Between Butter and
Margarine? Available at southernliving.com/fats/butter/difference-
between-butter-margarine.
2. Prättälä R, Levälahti E, Lallukka T, et al. From margarine to butter:
predictors of changing bread spread in an 11-year population follow-up.
Public Health Nutr. 2016;19(9):1707-17.

NUTRITIONAL FACTS
The Table below contains detailed information on the different
nutrients in butter (1).
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TABLE. NUTRITIONAL FACTS

Calorie Information
Amounts Per Selected Serving %DV
Calories 1628 (6816 kJ) 81%
From Carbohydrate 0.9 (3.8 kJ)
From Fat 1618 (6774 kJ)
From Protein 8.2 (34.3 kJ)
From Alcohol 0.0 (0.0 kJ)

Carbohydrates
Amounts Per Selected Serving %DV
Total Carbohydrate 0.1 g 0%
Dietary Fiber 0.0 g 0%
Starch 0.0 g
Sugars 0.1 g

Protein & Amino Acids

Amounts Per Selected Serving %DV
Protein 1.9 g 4%

Vitamins
Amounts Per Selected Serving %DV
Vitamin A 5673 IU 113%
Vitamin C 0.0 mg 0%
Vitamin D 127 IU 32%
Vitamin E (Alpha Tocopherol) 5.3 mg 26%
Vitamin K 15.9 mcg 20%
Thiamin 0.0 mg 1%
Riboflavin 0.1 mg 5%
Niacin 0.1 mg 0%
Vitamin B6 0.0 mg 0%
Folate 6.8 mcg 2%
Vitamin B12 0.4 mcg 6%
Pantothenic Acid 0.2 mg 2%
Choline 42.7 mg
Betaine 0.7 mg
 23

Ben-Nun L butter

Minerals
Amounts per Selected Serving %DV
Calcium 54.5 mg 5%
Iron 0.0 mg 0%
Magnesium 4.5 mg 1%
Phosphorus 54.5 mg 5%
Potassium 54.5 mg 2%
Sodium 1307 mg 54%
Zinc 0.2 mg 1%
Copper 0.0 mg 0%
Manganese 0.0 mg 0%
Selenium 2.3 mcg 3%
Fluoride 6.4 mcg

Sterols
Amounts Per Selected Serving %DV
Cholesterol 488 mg 163%
Phytosterols ~

Other
Amounts Per Selected Serving %DV
Alcohol 0.0 g
Water 36.0 g
Ash 4.8 g
Caffeine 0.0 mg
Theobromine 0.0 mg
Footnotes for Butter, salted
Source: Nutrient data for this listing was provided by USDA SR-21. "~"
indicates a missing or incomplete value. Percent Daily Values (%DV) are
for adults or children aged 4 or older, and are based on a 2,000 calorie
reference diet. His/her daily values may be higher or lower based on his/her
individual needs.
Nutrition Data's Opinion, Completeness Score™, Fullness Factor™,
Rating, Estimated Glycemic Load (eGL), and Better Choices Substitutions™
are editorial opinions of NutritionData.com, given without warranty, and are
not intended to replace the advice of a nutritionist or health-care
professional. Nutrition Data's opinions and ratings are based on weighted
averages of the nutrient densities of those nutrients for which the FDA has
established Daily Values, and do not consider other nutrients that may be
important to his/her health or take into account his/her individual needs.
 24

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Consequently, Nutrition Data's higher-rated foods may not necessarily be

healthier for you than lower-rated ones. All foods, regardless of their rating,
have the potential to play an important role in your diet.

Reference
1. Butter, salted Nutrition Facts & Calories. Available at
nutritiondata.self.com/facts/dairy-and-egg-products/0/2#ixzz5k0Axv 7xd.

CHANGING COMPOSITION
Bobe & al. (1) mentioned that changing the milk fatty acid
composition can improve the nutritional and physical properties of
dairy products and their acceptability to consumers. A more
healthful milk fatty acid composition can be achieved by altering the
cow's diet, for example, by feeding supplemental fish oil (FO) or
roasted soybeans (RSB), or by selecting cows with a more
unsaturated milk fatty acid composition. The authors (1) examined
whether feeding supplemental FO or RSB to cows that had a more
unsaturated milk fatty acid composition acted additively to produce
butter with improved fatty acid composition and texture. Using a 3 x
3 Latin square design with 2 replications, diets to multiparous
Holstein cows (60 to 200 DIM) were chosen for producing either
more or less unsaturated milk fatty acid composition (n=6 for each
group) for three 3-wk periods. The control diet contained 3.7% crude
fat and the 2 experimental diets contained, on a dry matter basis,
0.8% of additional lipids in the form of 0.9% of FO or 5% of RSB. The
milk, collected in the third week of feeding, was used to make butter,
which was analyzed for its fatty acid composition and physical
properties. Dry matter intake, milk yield, and milk composition were
not significantly affected by cow diet or by cow selection. Cows that
produced a more unsaturated and healthful milk fat prior to the
feeding study, according to a "health-promoting index" [HPI = (sum of
% of unsaturated fatty acids)/ (%12:0 + 4 x %14:0 + %16:0)],
maintained a higher HPI in their butter during the feeding study than
did cows with a low HPI. Milk from cows fed supplemental FO or RSB
yielded more unsaturated butters with a higher HPI. This butter also
was softer when the cows were fed RSB. Feeding RSB to cows
chosen for their high milk HPI yielded the most unsaturated butter
with the highest HPI and softest texture. Thus, selecting cows with a
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more health-promoting milk fatty acid composition and feeding

supplemental RSB can be used in combination to produce butter that
has a consumer-friendly texture and a healthful fatty acid profile (1).
Schripsema (2) revealed that the separation of butter or
margarine into polar (soluble in water) and apolar fractions (soluble
in chloroform) and subsequent analysis of these fractions by (1)H
NMR permits a comprehensive analysis of its constituents. In the
polar fraction the preservatives benzoic and sorbic acid, the organic
acids citric, lactic, butyric, acetic, and formic acid, and, furthermore,
the carbohydrate lactose were quantified. In the apolar fraction the
conjugated linoleic acid (CLA) rumenic acid, diglycerides, and linoleic
acid were quantified. Rumenic acid is a characteristic component of
ruminant fats and was found in all butter samples. The levels varied
between 0.50 and 1.08%. Ten brands of Brazilian butter were
investigated as was one brand from Norway. Also, two brands of
margarine were investigated for comparison. A large variation in
especially polar constituents was found between the butter samples,
revealing the presence of preservatives in five brands of butter from
Brazil, remarkable because these additives are legally not allowed.
Furthermore, the levels of organic acids and lactose permitted
conclusions about the production process and quality; for example,
the presence of higher levels of free butyric acid indicate lipolysis,
leading to a lower quality, and low levels of lactose indicate that after
churning the residual milk fluids have been removed by an additional
washing step in the production process (2).
Honfo & al. (3) noticed that increasing demand of shea products
(kernels and butter) has led to the assessment of the state-of-the-art
of these products. In this review, attention has been focused on
macronutrients and micronutrients of pulp, kernels, and butter of
shea tree and also the physicochemical properties of shea butter.
Surveying the literature revealed that the pulp is rich in vitamin C
(196.1 mg/100 g); consumption of 50 g covers 332% and 98% of the
recommended daily intake (RDI) of children (4-8 years old) and
pregnant women, respectively. The kernels contain a high level of fat
(17.4-59.1 g/100 g dry weight). Fat extraction is mainly done by
traditional methods that involve roasting and pressing of the kernels,
churning the obtained liquid with water, boiling, sieving, and cooling.
The fat (butter) is used in food preparation and medicinal and
cosmetics industries. Its biochemical properties indicate some
 26

Ben-Nun L butter

antioxidant and anti-inflammatory activities. Large variations are

observed in the reported values for the composition of shea
products. Recommendations for future research are presented to
improve the quality and the shelf-life of the butter. In addition, more
attention should be given to the accuracy and precision in
experimental analyses to obtain more reliable information about
biological variation (3).
Verardo & al. (4) noticed bovine milk contains hundreds of
diverse components, including proteins, peptides, amino acids, lipids,
lactose, vitamins and minerals. Specifically, the lipid composition is
influenced by different variables such as breed, feed and
technological process. In this study the fatty acid and phospholipid
compositions of different samples of butter and its by-products from
the Parmigiano Reggiano cheese area, produced by industrial and
traditional churning processes, were determined. The fatty acid
composition of samples manufactured by the traditional method
showed higher levels of monounsaturated and polyunsaturated fatty
acids compared with industrial samples. In particular, the contents of
n-3 fatty acids and conjugated linoleic acids were higher in samples
produced by the traditional method than in samples produced
industrially. Sample phospholipid composition also varied between
the two technological processes. Phosphatidyl ethanolamine was the
major phospholipid in cream, butter and buttermilk samples
obtained by the industrial process as well as in cream and buttermilk
samples from the traditional process, while phosphatidylcholine was
the major phospholipid in traditionally produced butter. This result
may be explained by the different churning processes causing
different types of membrane disruption. Generally, samples
produced traditionally had higher contents of total phospholipids; in
particular, butter produced by the traditional method had total
phospholipid content 33% higher than that of industrially produced
butter. The data show that the samples studied represent the two
types of products present in the Parmigiano Reggiano cheese area,
where the industrial churning process is widespread compared with
the traditional processing of Reggiana cow's milk. This is because
Reggiana cow's milk production is lower than that of other breeds
and the traditional churning process is time-consuming and
economically disadvantageous. However, its products have been
demonstrated to contain more bioactive lipids compared with
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products obtained from other breeds and by the industrial process

(4).
Wendlinger & Vetter (5) noticed Furan fatty acids (F-acids) are
valuable antioxidants containing a furan moiety in the central part of
the molecule. They occur in the lipids of different foodstuffs and
plants, with grass being the main source for their presence in milk fat
and butter. Because cows from organic farming receive higher
portions of grass-based feed, it was tested whether organic butter
samples (n=26) contain more F-acids than conventional ones (n=25)
in Germany. For this purpose, samples were melted, and the lipid
phase was separated and transesterified into methyl esters, which
were enriched using silver ion chromatography and analyzed by GC-
EI/MS in the selected ion monitoring (SIM) mode. Levels of F-acids in
butter were higher in summer than in winter, and in both seasons,
organic samples contained significantly higher levels of F-acids than
conventional ones (one-way ANOVA: p<0.001). The daily intake of F-
acids via milk fat and other foodstuffs was calculated (5).
Sloffer & al. (6) mentioned that lipid-based nutrient supplements
(LNS) are used to prevent and treat moderate and severe acute
malnutrition, a leading cause of mortality in children-under-five. The
physical and chemical changes of two new LNS products were
evaluated before and after accelerated shelf life testing (ASLT)
according to protocols suggested by the U.S. Agency for
International Development (USAID) and Doctors without Borders and
compared against USAID's A-20 paste as a control. LNS formulas
containing Shea butter from the Shea nut tree (Vitellaria paradoxa), a
common fat source in parts of Sub-Saharan Africa, with and without
flax-seed oil, as a source of omega-3 fatty acids, were developed. LNS
formulas were batched (0.8 kg) in a wet grinder, sealed under
nitrogen in three-layer mini-pouches (20 g), and underwent ASLT at
40 ± 2°C for six months with sampling every eight weeks. At each
time point, water activity, moisture, peroxide value, oil separation,
vitamin C content, and hardness were evaluated. Results showed
comparable stability among all formulas with an increase in Aw
(p<0.05) but no change in vitamin C, oil separation, or peroxide value.
Addition of Shea butter improved the LNS's hardness, which
remained stable over time. Modifying fat profile in LNS can improve
its texture and essential fatty acid content without affecting its
storage stability (6).
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Ben-Nun L butter

Geithe & al. (7) revealed that with ∼400 olfactory G protein-
coupled receptors (GPCR), humans sensitively perceive ∼230 key
aroma compounds as best natural agonists of ∼10,000 food volatiles.
An understanding of odorant coding, thus, critically depends on the
knowledge about interactions of key food aroma chemicals and their
mixtures with their cognate receptors. Genetically designed test cell
systems enable the screening, deorphaning, and characterization of
single odorant receptors (OR). This study shows for the food aroma-
specific and quantitative butter aroma recombinate, and its single
components, specific in vitro class-I OR activity patterns, as well as
the activation of selected OR in a concentration-dependent manner.
Recently, chemosensory receptors, especially class-I OR, were
demonstrated to be expressed on blood leukocytes, which may
encounter foodborne aroma compounds postprandially. Butter
aroma recombinate induced chemotaxis of isolated human
neutrophils in a defined gradient, and in a concentration-dependent
and pertussis toxin-sensitive manner, suggesting at least a GPCR-
mediated activation of blood leukocytes by key food odorants (7).
Botta & Ghosh (8) stated that anecdotal evidence suggests that
the incorporation of n-6 polyunsaturated fatty acid (n-6 PUFA)
containing oilseeds in dairy feeds depletes saturated fatty acids (SFA)
in dairy fats such as butter. However, due to the lack of chemical
evidence, the current status of n-6 PUFA or SFA in butter is unknown.
It was hypothesized that n-6 PUFA levels in commercial butter were
inversely proportional to its SFA content and directly proportional to
the extent of n-6 PUFA-rich oilseed production of its country of
origin. Grass-fed and commercial butters from Australia, Belarus,
Canada, China, England, France, Germany, Iceland, India, Israel,
Japan, the Netherlands, New Zealand, Russia, and the United States
were analyzed via gas chromatography. Extent of n-6 PUFA
containing oilseed production for countries was obtained from the
FAOStat 2015 database. Globally, SFA from commercial butters had a
strong negative correlation (Spearman r = -0.53, p = 0.025) with its n-
6 PUFA content, with U.S. and Canadian butter demonstrating the
highest n-6 PUFA as well as n-6/n-3 PUFA ratios. As predicted, we
show that countries with >5% of its agricultural land dedicated to n-6
PUFA oilseed production demonstrate a "spillover" increase of n-6
PUFA in their commercial butters (Spearman r = 0.85, p=0.0054). The
overall significance of this study is that it presents novel evidence of
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the global impact of rising n-6 PUFA production on commercial butter

fat composition. The authors hope these data will lead to inclusion of
actual biochemical analyses of dairy fats in future clinical trials. This
inclusion of analyses will better explain the differential health
outcomes among different countries for such interventions (8).
Kalinin & al. (9) reported that in organism, differences in physical
chemical properties and physiological role of positional isomeric
forms of triglycerides of fatty acids consumed with food are well
known. The Raman spectroscopy to analyze there is possible to be
applied. Both of these circumstances are to be considered and
applied in resolving practical issues of dietetics, e.g., in detection of
falsification and replacement of butter and spreads with palm oil, as
well as in monitoring ratio of fatty consumed by patients with
cardiologic and oncologic pathology. The purpose of study is to
determine possibilities of analyzing positional fatty acids isomers
specific for butter and palm oil using portable and operational
spectrometers of absorption in near infrared range. The technique of
projections on latent structures was applied to obtain calibrations of
spectrometers to detect content of 7 regulated fatty acids and
percentage of milk fat and palm oil in fat-oil mixtures according
specters of Raman and near infrared range spectrometers. The
possibility of identifying positional isomers of animal and vegetable
triglycerides according Raman spectra was confirmed. Besides, it is
established that efficiency of determining (reliability, accuracy and
selectivity) the proportions of the above oils using both Raman
spectra and near-infrared optical density spectra was much higher
than when calculating the percentage of the same oils using the
content of 7 fatty acids. This fact reflects sensitivity of near infrared
range absorption spectra both to length of the carbon chain and
degree of unsaturation (number of double bonds C=C) of fatty acids
and positional isomeric forms of triglycerides. The obtained data are
used in formulating technical requirements and conditions for
application of portable near infrared range spectrometer for mass
analysis of fat-oil products (9).
Muzhingi & al. (10) mentioned that Kale is a rich source of
provitamin A- β-carotene. This study used intrinsically labeled kale
*2H9+ β-carotene to determine the effect of peanut butter on the
bioconversion of kale β-carotene to vitamin A in preschool children.
Preschool children (n=37; age 12-36 months) were randomly
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assigned to 50 g cooked kale (1.5 mg β-carotene content) with either

33 g peanut butter (PBG) or with 16 g lard (LG) and a reference dose
of 1 mg [13C10] retinyl acetate capsule. Blood samples were
processed to serum and analyzed by Negative Chemical Ionization-
Gas Chromatography Mass Spectrometry (NCI-GCMS) for the
enrichments of *2H+ retinol from kale *2H9+ β-carotene and [13C10]
retinol from reference dose. The area under curves (AUCs) of molar
enrichment at days 1, 2, 3, 6, 15, and 21 after the labeled doses was
56.3±10.5 and 84.8±16.2 (nmole) for [2H] retinol from LG and PBG
kale *2H9+ β-carotene, respectively. The AUC of [13C10] retinol from
reference dose was 432.6±54.9 (LG) and 560.3±156.7 (nmole) (PBG),
respectively. The calculated β-carotene conversion factors were
13.4±3.1 and 11.0±3.9 to 1 (p>0.05) by weight for LG and PBG,
respectively. This study showed that peanut butter enhances the
vitamin A value of kale (10).

Bread and Butter

James Del Grosso

Gezaheg & al. (11) mentioned that the quality of Shea butter is
highly affected by processing factors. Hence, the aim of this work
was to evaluate the effects of conditioning duration (CD), moisture
content (MC), and die temperature (DT) of screw expeller on Shea
butter quality. A combination of 33 full factorial design and response
surface methodology was used for this investigation. Response
variables were refractive index, acid value, and peroxide value. The
model enabled to identify the optimum operating settings (CD = 28-
30 min, MC = 3-5 g/100 g, and DT = 65-70°C) for maximize refractive
index and minimum acid value. For minimum peroxide value 0 min
CD, 10 g/100 g MC, and 30°C were discovered. In all-over
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Ben-Nun L butter

optimization, optimal values of 30 min CD, 9.7 g/100 g MC, and 70°C
DT were found. Hence, the processing factors must be at their
optimal values to achieve high butter quality and consistence (11).

This chapter (1-11) deals with a different composition and texture

of a variety of butters.

References
1. Bobe G, Zimmerman S, Hammond EG, et al. Butter composition and
texture from cows with different milk fatty acid compositions fed fish oil
or roasted soybeans. J Dairy Sci. 2007;90(6):2596-603.
2. Schripsema J. Comprehensive analysis of polar and apolar
constituents of butter and margarine by nuclear magnetic resonance,
reflecting quality and production processes. J Agric Food Chem. 2008;
56(8):2547-52.
3. Honfo FG, Akissoe N, Linnemann AR, et al. Nutritional composition
of shea products and chemical properties of shea butter: a review. Crit
Rev Food Sci Nutr. 2014;54(5):673-86.
4. Verardo V, Gómez-Caravaca AM, Gori A, et al. Bioactive lipids in the
butter production chain from Parmigiano Reggiano cheese area. J Sci Food
Agric. 2013;93(14):3625-33.
5. Wendlinger C, Vetter W. High concentrations of furan fatty acids in
organic butter samples from the German market. J Agric Food Chem.
2014;62(34):8740-4.
6. Sloffer EM, Gaur S, Engeseth NJ, Andrade JE. Development and
physico-chemical characterization of a shea butter-containing lipid
nutrition supplement for Sub-Saharan Africa. Foods. 2017 Nov 8;6(11).
pii: E97.
7. Geithe C, Andersen G, Malki A, Krautwurst D. A butter aroma
recombinate activates human class-I odorant receptors. J Agric Food
Chem. 2015;63(43):9410-20.
8. Botta A, Ghosh S. Exploring the impact of n-6 PUFA-rich oilseed
production on commercial butter compositions worldwide. J Agric Food
Chem. 2016;64(42):8026-34.
9. Kalinin AV, Krasheninnikov VN, Titov VN. The spectrometry of
isomers of triglycerides of fatty acids in fatty buttery products: butter and
palm oil. Klin Lab Diagn. 2018;63(5):260-7.
10. Muzhingi T, Yeum KJ, Bermudez O, et al. Peanut butter increases
the bioavailability and bioconversion of kale β-carotene to vitamin A. Asia
Pac J Clin Nutr. 2017;26(6):1039-47.
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11. Gezahegn YA, Emire SA, Asfaw SF. Optimization of Shea (Vitellaria
paradoxa) butter quality using screw expeller extraction. Food Sci Nutr.
2016;4(6):840-847. eCollection 2016 Nov.

CONSUMPTION
Continent-wise, Europe dominates consumption of butter while
Africa consumes the least. Surprisingly, Oceania has just two
countries listed that consume butter, New Zealand and Australia, but
their consumption easily beats that of other continents such as Africa
and the Middle East quite easily (1).
The top consumer of butter in the world is France with a per
capita butter consumption of 8.2 kg. The second nation is Denmark
with a per capita consumption of 6.4 kg, and it is followed closely by
Iceland with 6kg. In the fourth position is Czech Republic with 5.4 kg
while Switzerland closes the top five with 5.2 kg. The only other
country with a per capita consumption of above 5 kg is New Zealand
in the sixth position. Frances domination in this list is
unquestionable. Compared to the second country, Denmark, France
has a higher consumption margin of 1.8 kg. Compared to the last five
nations (Belarus, Finland, Lithuania, India, Slovakia, and Australia),
France has at least twice as much consumption as each of these
countries. In previous years, up to 2014 the per capita consumption
of butter by France been the same or higher. The highest recorded
consumption was in 2014 with a consumption of 8.4 kg. Denmark’s
consumption has also been steadily increasing all the way from 2012.
In fact, Denmark’s increased consumption saw it leapfrog Iceland into
second place in 2016. In 2016, Denmark has a recorded consumption
of 5.0 kg while Iceland had 5.8 kg. However, this is not to say that
Iceland’s consumption decreased. On the contrary, Iceland had a
growth of 0.2kg. Denmark’s growth was by 1.4 kg. The last country,
Finland has had mixed consumption levels in previous years. Looking
at the other end of the spectrum, Finland is the last in the top fifteen
ranking with a per capita consumption of just above 3 kg. Belarus
and Lithuania tie at position thirteen with a consumption of 3.8 kgs.
Interestingly, the same thing happens with India and Slovakia at
position eleven with a slightly higher consumption of 3.9 kg.
Australia has a 0.1 kg more consumption than India and Slovakia (1).
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Table. Countries Who Consume the Most Butter (1).

Rank Country Butter Consumption (Kg Per Capita)

1 France 8.2

2 Denmark 6.4

3 Iceland 6.0

4 Czech Republic 5.4

5 Switzerland 5.2

6 New Zealand 5.1

8 Pakistan 4.8

9 Poland 4.5

10 Australia 4.0

11 Slovakia 3.9

12 India 3.9

13 Lithuania 3.8

14 Belarus 3.8

15 Finland 3.3

Krause & al. (2) identified and explored the sensory

characteristics that drive consumer liking of butter. A trained
descriptive panel evaluated 27 commercial butters using a defined
sensory language. Two focus groups were conducted with butter
consumers to gain an understanding of consumer use and
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consumption habits. Six representative butters and 2 vegetable oil

spreads were selected for consumer acceptance testing. Both
internal and external preference mapping techniques were applied to
interpret consumer data. Key discriminating sensory characteristics
of butters included color intensity; diacetyl, cooked, grassy, and milk
fat flavors; and salty taste. From focus groups and quantitative
consumer testing, the key butter features were a desirable flavor and
a natural image. Negative aspects included price and cholesterol.
Five consumer clusters with distinct butter and spread flavor likes
and dislikes were identified. Butter is a desirable product to
consumers. Sensory expectations of butter vary among consumers,
and butters with specific sensory characteristics could be marketed
to specific target market segments (2).

References
1. Countries Who Consume the Most Butter. Available at
worldatlas.com/articles/countries-who-consume-the-most-butter.html.
2. Krause AJ, Lopetcharat K, Drake MA. Identification of the
characteristics that drive consumer liking of butter. J Dairy Sci. 2007;
90(5):2091-102.

BUTTER ADULTERATION
Mabood & al. (1) reported that Cows' butterfat may be
adulterated with animal fat materials like tallow which causes
increased serum cholesterol and triglycerides levels upon
consumption. There is no reliable technique to detect and quantify
tallow adulteration in butter samples in a feasible way. In this study,
a highly sensitive Near-infrared (NIR) spectroscopy combined with
chemometric methods was developed to detect as well as quantify
the level of tallow adulterant in clarified butter samples. For this
investigation, the pure clarified butter samples were intentionally
adulterated with tallow at the following percentage levels: 1%, 3%,
5%, 7%, 9%, 11%, 13%, 15%, 17% and 20% (wt/wt). Altogether 99
clarified butter samples were used including nine pure samples (un-
adulterated clarified butter) and 90 clarified butter samples
adulterated with tallow. Each sample was analyzed by using NIR
spectroscopy in the reflection mode in the range 10,000-4000 cm-1,
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at 2 cm-1 resolution and using the transflectance sample accessory

which provided a total path length of 0.5 mm. Chemometric models
including principal components analysis (PCA), partial least-squares
discriminant analysis (PLSDA), and partial least-squares regressions
(PLSR) were applied for statistical treatment of the obtained NIR
spectral data. The PLSDA model was employed to differentiate pure
butter samples from those adulterated with tallow. The employed
model was then externally cross-validated by using a test set which
included 30% of the total butter samples. The excellent performance
of the model was proved by the low RMSEP value of 1.537% and the
high correlation factor of 0.95. This newly developed method is
robust, non-destructive, highly sensitive, and economical with very
minor sample preparation and good ability to quantify less than 1.5%
of tallow adulteration in clarified butter samples (1).
Lohumi & al. (2) mentioned that spectroscopic techniques for
food quality analysis are limited to surface inspections and are highly
affected by the superficial layers (skin or packaging material) of the
food samples. The ability of spatially offset Raman spectroscopy
(SORS) to obtain chemical information from below the surface of a
sample makes it a promising candidate for the non-destructive
analysis of the quality of packaged food. In the present study, a line-
scan SORS technique was developed for obtaining the Raman spectra
of packaged-food samples. This technique was used to quantify
butter adulteration with margarine through two different types of
packaging. Further, the significant commercial potential of the
developed technique was demonstrated by its being able to
discriminate between ten commercial varieties of butter and
margarine whilst still in their original, unopened packaging. The
results revealed that, while conventional backscattering Raman
spectroscopy cannot penetrate the packaging, thus preventing its
application to the quality analysis of packaged food, SORS analysis
yielded excellent qualitative and quantitative analyses of butter
samples. The partial least-square regression analysis predictive
values for the SORS data exhibit correlation coefficient values of 0.95
and 0.92, associated with the prediction error 3.2% and 3.9% for
cover-1 & 2, respectively. The developed system utilizes a laser line
(ca. 14-cm wide) that enables the simultaneous collection of a large
number of spectra from a sample. Thus, by averaging the spectra
collected for a given sample, the signal-to-noise ratio of the final
 36

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spectrum can be enhanced, which will then have a significant effect

on the multivariate data analysis methods used for qualitative and/or
qualitative analyses. This recently presented line-scan SORS
technique could be applied to the development of high-throughput
and real-time analysis techniques for determining the quality and
authenticity various packaged agricultural products (2).
Kim & al. (3) assessed the potential application of gas
chromatography (GC) in detecting Milk Fat (MF) adulteration with
vegetable oils and animal fats and of characterizing samples by fat
source. One hundred percent pure MF was adulterated with
different vegetable oils and animal fats at various concentrations
(0%, 10%, 30%, 50%, 70%, and 90%). GC was used to obtain the fatty
acid (FA) profiles, triacylglycerol (TG) contents, and cholesterol
contents. The pure MF and the adulterated MF samples were
discriminated based on the total concentrations of saturated FAs and
on the 2 major FAs (oleic acid [C18:1n9c] and linoleic acid [C18:2n6c],
TGs [C52 and C54], and cholesterol contents using statistical analysis
to compared difference. These bio-markers enabled the detection of
as low as 10% adulteration of non-MF into 100% pure MF. The study
demonstrated the high potential of GC to rapidly detect MF
adulteration with vegetable and animal fats, and discriminate among
commercial butter and milk products according to the fat source.
These data can be potentially useful in detecting foreign fats in these
butter products. It is important to consider that several individual
samples should be analyzed before coming to a conclusion about MF
authenticity (3).
Fadzillah & al. (4) stated that the authentication of food products
from the presence of non-allowed components for certain religion
like lard is very important. In this study, proton Nuclear Magnetic
Resonance ((1)H-NMR) spectroscopy was used for the analysis of
butter adulterated with lard by simultaneously quantification of all
proton bearing compounds, and consequently all relevant sample
classes. Since the spectra obtained were too complex to be analyzed
visually by the naked eyes, the classification of spectra was carried
out. The multivariate calibration of Partial Least Square (PLS)
regression was used for modeling the relationship between actual
value of lard and predicted value. The model yielded a highest
regression coefficient (R(2)) of 0.998 and the lowest root mean
square error calibration (RMSEC) of 0.0091% and root mean square
 37

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error prediction (RMSEP) of 0.0090, respectively. Cross validation

testing evaluates the predictive power of the model. PLS model was
shown as good models as the intercept of R(2)Y and Q(2)Y were
0.0853 and -0.309, respectively (4).

This chapter (1-4) demonstrates cows' butterfat may be

adulterated with animal fat materials like tallow which causes
increased serum cholesterol and triglycerides levels upon
consumption.
There are various methods that can detect butter adulteration.

References
1. Mabood F, Abbas G, Jabeen F, et al. Robust new NIRS coupled with
multivariate methods for the detection and quantification of tallow
adulteration in clarified butter samples. Food Addit Contam Part A Chem
Anal Control Expo Risk Assess. 2018;35(3):404-1.
2. Lohumi S, Lee H, Kim MS, e al. Through-packaging analysis of butter
adulteration using line-scan spatially offset Raman spectroscopy. Anal
Bioanal Chem. 2018;410(22):5663-73.
3. Kim JM, Kim HJ, Park JM. Determination of milk fat adulteration
with vegetable oils and animal fats by gas chromatographic analysis. J
Food Sci. 2015;80(9):C1945-51.
4. Fadzillah NA, Man Yb, Rohman A, et al. Detection of butter
adulteration with lard by employing (1)H-NMR spectroscopy and
multivariate data analysis. J Oleo Sci. 2015;64(7):697-703.

HEALTH EFFECTS
METABOLIC SYNDROME
Hosseinpour-Niazi & al. (1) investigated the association between
Hydrogenated- (HVOs) and non-Hydrogenated Vegetable Oils (non-
HVOs) and butter and the Metabolic Syndrome (MetS) after 3-years
of follow-up in adults. This study was conducted between 2006-2008
and 2009-2011 within the framework of the Tehran Lipid and Glucose
Study, on 1,582 adults, aged 19-84 years. Intakes of HVOs, non-HVOs
and butter were assessed by a validated semi-quantitative food
frequency questionnaire. Based on the consumption of food rich in
fat including HVOs, non-HVOs and butter, participants were
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categorized to consumers and non-consumers. Of 1,582 participants

during a 3-year follow-up, 15.2% developed MetS. Non-consumption
of butter was associated with lower MetS risk compared with its
consumption. Among consumers of food rich in fat, intake of HVOs
and butter were associated with an increased risk of MetS; ORs in the
final multivariate model were 2.70 (95% CI 1.52-4.78) for HVOs and
2.03 (95% CI 1.20-3.41) for butter, in the highest, compared to the
lowest category of dietary intakes. Intake of non-HVOs was not
associated with risk of MetS. The data show that consumption of
HVOs and butter were positively associated with an increased risk of
MetS (1).
Werner & al. (2) reported that there is considerable interest in
dairy products from low-input systems, such as mountain-pasture
grazing cows, because these products are believed to be healthier
than products from high-input conventional systems. This may be
due to a higher content of bioactive components, such as phytanic
acid, a PPAR-agonist derived from chlorophyll. However, the effects
of such products on human health have been poorly investigated.
This study compared the effect of milk-fat from mountain-pasture
grazing cows (G) and conventionally fed cows (C) on risk markers of
the MetS. In a double-blind, randomized, 12-week, parallel
intervention study, 38 healthy subjects replaced part of their habitual
dietary fat intake with 39 g fat from test butter made from milk from
mountain-pasture grazing cows or from cows fed conventional winter
fodder. Glucose-tolerance and circulating risk markers were analyzed
before and after the intervention. No differences in blood lipids,
lipoproteins, hsCRP, insulin, glucose or glucose-tolerance were
observed. Interestingly, strong correlations between phytanic acid at
baseline and total (p<0.0001) and Low Density Lipoprotein
Cholesterol (LDL-C) (p=0.0001) were observed. The data
demonstrate that lack of effects on blood lipids and inflammation
indicates that dairy products from mountain-pasture grazing cows
are not healthier than products from high-input conventional
systems. Considering the strong correlation between LDL-C and
phytanic acid at baseline, it may be suggested that phytanic acid
increases total and LDL-C (2).

This chapter (1-2) shows that consumption of HVOs and butter

are associated with an increased risk of MetS.
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Dairy products from mountain-pasture grazing cows are not

healthier than products from high-input conventional systems.

References
1. Hosseinpour-Niazi S, Mirmiran P, Hosseini-Esfahani F, Azizi F. Is the
metabolic syndrome inversely associates with butter, non-hydrogenated-
and hydrogenated-vegetable oils consumption: Tehran lipid and glucose
study. Diabetes Res Clin Pract. 2016;112:20-9.
2. Werner LB, Hellgren LI, Raff M, et al. Effects of butter from
mountain-pasture grazing cows on risk markers of the metabolic syndrome
compared with conventional Danish butter: a randomized controlled
study. Lipids Health Dis. 2013 Jul 10;12:99.

LIPOPROTEINS. Engel & Tholstrup (1) noticed that butter is

known to have a cholesterol-raising effect and, therefore, has often
been included as a negative control in dietary studies, whereas the
effect of moderate butter intake has not been elucidated to the
knowledge. The effects of moderate butter intake, moderate olive oil
intake, and a habitual diet were compared on blood lipids, High-
Sensitivity C-reactive Protein (hsCRP), glucose, and insulin. The study
was a controlled, double-blinded, randomized 2 × 5-wk crossover
dietary intervention study with a 14-d run-in period during which
subjects consumed their habitual diets. The study included 47
healthy men and women (mean ± SD total cholesterol (TC): 5.22 ±
0.90 mmol/L) who substituted a part of their habitual diets with 4.5%
of energy from butter or refined olive oil. Study subjects were 70%
women with a mean age and body mass index (kg/m²) of 40.4 y and
23.5, respectively. Butter intake increased TC and low density
lipoprotein cholesterol (LDL-C) more than did olive oil intake (p<0.05)
and the run-in period (p<0.005 and p<0.05, respectively) and
increased high density lipoprotein cholesterol (HDL-C) compared with
the run-in period (p<0.05). No difference in effects was observed for
triacylglycerol, hsCRP, insulin, and glucose concentrations. The
intake of saturated fatty acids was significantly higher in the butter
period than in the olive oil and run-in periods (p<0.0001). Moderate
intake of butter resulted in increases in TC and LDL-C compared with
the effects of olive oil intake and a habitual diet (run-in period).
Furthermore, moderate butter intake was also followed by an
increase in HDL-C compared with the habitual diet. The data show
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that hypercholesterolemic people should keep their consumption of

butter to a minimum, whereas moderate butter intake may be
considered part of the diet in the normocholesterolemic population
(1).
Brassard & al. (2) noticed that controversies persist concerning
the association between intake of dietary saturated fatty acids (SFAs)
and cardiovascular disease risk. The impact of consuming equal
amounts of SFAs from cheese and butter were compared on
cardiometabolic risk factors. In a multicenter, crossover, randomized
controlled trial, 92 men and women with abdominal obesity and
relatively low HDL-C concentrations were assigned to sequences of 5
predetermined isoenergetic diets of 4 weeks each separated by 4-
week washouts: 2 diets rich in SFAs (12.4-12.6% of calories) from
either cheese or butter; a monounsaturated fatty acid (MUFA)-rich
diet (SFAs: 5.8%, MUFAs: 19.6%); a polyunsaturated fatty acid
(PUFA)-rich diet (SFAs: 5.8%, PUFAs: 11.5%); and a low-fat, high-
carbohydrate diet (fat: 25%, SFAs: 5.8%). Serum HDL-C
concentrations were similar after the cheese and butter diets but
were significantly higher than after the carbohydrate diet (+3.8% and
+4.7%, respectively; p< 0.05 for both). LDL-C concentrations after the
cheese diet were lower than after the butter diet (-3.3%, p<0.05) but
were higher than after the carbohydrate (+2.6%), MUFA (+5.3%), and
PUFA (+12.3%) diets (p<0.05 for all). LDL-C concentrations after the
butter diet also increased significantly (from +6.1% to +16.2%,
p<0.05) compared with the carbohydrate, MUFA, and PUFA diets.
The LDL-C response to treatment was significantly modified by
baseline values (p-interaction = 0.02), with the increase in LDL-C
being significantly greater with butter than with cheese only among
individuals with high baseline LDL-C concentrations. There was no
significant difference between all diets on inflammation markers,
blood pressure, and insulin-glucose homeostasis. The results of this
study suggest that the consumption of SFAs from cheese and butter
has similar effects on HDL-C but differentially modifies LDL-C
concentrations compared with the effects of carbohydrates, MUFAs,
and PUFAs, particularly in individuals with high LDL-C. In contrast,
SFAs from either cheese or butter have no significant effects on
several other nonlipid cardiometabolic risk factors (2).
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Motte de Beurre
Antoine Vollon. c.1880, National Gallery of Art

Zock & Katan (3) mentioned that intake of trans fatty acids
unfavorably affects blood lipoproteins. As margarines are a major
source of trans, claims for the advantages of margarines over butter
need to be scrutinized. Here dietary trials that directly compared the
effects of butter and margarine on blood lipids were reviewed.
Twenty studies in which subjects had stable body weights, and
margarine and butter were exchanged in the diet at constant energy
and fat intake were identified. The changes in average blood lipid
levels between study diets (49 comparisons) as a function of the
percentage of calories as margarine substituted for butter were
calculated. Replacing 10% of calories from butter by hard high-trans
stick margarines lowered TC by 0.19, LDL by 0.11, and HDL by 0.02
mmol/l, and did not affect the total/HDL–C ratio. Soft low-trans tub
margarines decreased TC by 0.25 and LDL by 0.20 mmol/l, did not
affect HDL, and decreased the TC/HDL-C ratio by 0.20. Based on the
total/HDL-C ratio, replacement of 30 g of butter per day by soft tub
margarines would theoretically predict a reduction in coronary heart
disease (CHD) risk of 10%, while replacement of butter by hard, high-
trans margarines would have no effect. Replacing butter by low-
trans soft margarines favorably affects the blood lipoprotein profile
and may reduce the predicted risk of CHD, but high-trans hard
margarines probably confer no benefit over butter (3).
Denke (4) stated that it has been known for some time that cocoa
butter, although rich in saturated fatty acids, does not raise total
serum cholesterol concentrations as much as expected from its total
saturated fatty acid content. Whether the effect of cocoa butter
feeding on LDL-C concentrations was also less than predicted by its
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total saturated fatty acid content needed to be tested. In a recent

experiment cocoa butter did not raise LDL-C as much as predicted by
its total saturated fatty acid content. However, because of its
significant palmitic acid content, cocoa butter did raise LDL-C
concentrations more than do most liquid vegetable oils (4).
Sharma & al. (5) mentioned that Ghee, also known as clarified
butter, has been utilized for thousands of years in Ayurveda as a
therapeutic agent. In ancient India, ghee was the preferred cooking
oil. In the last several decades, ghee has been implicated in the
increased prevalence of coronary artery disease (CAD) in Asian
Indians due to its content of saturated fatty acids and cholesterol
and, in heated ghee, cholesterol oxidation products. The previous
research on Sprague-Dawley outbred rats, which serve as a model for
the general population, showed no effect of 5% and 10% ghee-
supplemented diets on serum cholesterol and triglycerides.
However, in Fischer inbred rats, which serve as a model for genetic
predisposition to diseases, results of the previous research showed
an increase in serum TC and triglyceride levels when fed a 10% ghee-
supplemented diet. In the present study, the effect of 10% dietary
ghee was investigated on microsomal lipid peroxidation, as well as
serum lipid levels in Fischer inbred rats to assess the effect of ghee
on free radical mediated processes that are implicated in many
chronic diseases including cardiovascular disease. Results showed
that 10% dietary ghee fed for 4 weeks did not have any significant
effect on levels of serum total cholesterol, but did increase
triglyceride levels in Fischer inbred rats. Ghee at a level of 10% in the
diet did not increase liver microsomal lipid peroxidation or liver
microsomal lipid peroxide levels. Animal studies have demonstrated
many beneficial effects of ghee, including dose-dependent decreases
in serum TC, LDL, Very Low Density Lipoprotein (VLDL), and
triglycerides; decreased liver TC, triglycerides, and cholesterol esters;
and a lower level of nonenzymatic-induced lipid peroxidation in liver
homogenate. Similar results were seen with heated (oxidized) ghee
which contains cholesterol oxidation products. A preliminary clinical
study showed that high doses of medicated ghee decreased serum
cholesterol, triglycerides, phospholipids, and cholesterol esters in
psoriasis patients. A study on a rural population in India revealed a
significantly lower prevalence of CHD in men who consumed higher
amounts of ghee. Research on Maharishi Amrit Kalash-4 (MAK-4), an
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Ayurvedic herbal mixture containing ghee, showed no effect on levels

of serum cholesterol, HDL, LDL, or triglycerides in hyperlipidemic
patients who ingested MAK-4 for 18 weeks. MAK-4 inhibited the
oxidation of LDL in these patients. The data available in the literature
do not support a conclusion of harmful effects of the moderate
consumption of ghee in the general population. Factors that may be
involved in the rise of CAD in Asian Indians include the increased use
of vanaspati (vegetable ghee) which contains 40% trans fatty acids,
psychosocial stress, insulin resistance, and altered dietary patterns.
Research findings in the literature support the beneficial effects of
ghee outlined in the ancient Ayurvedic texts and the therapeutic use
of ghee for thousands of years in the Ayurvedic system of medicine
(5).
Khaw & al. (6) stated that high dietary saturated fat intake is
associated with higher blood concentrations of LDL-C, an established
risk factor for CHD. However, there is increasing interest in whether
various dietary oils or fats with different fatty acid profiles such as
extra virgin coconut oil may have different metabolic effects but trials
have reported inconsistent results. This study (6) compared changes
in blood lipid profile, weight, fat distribution and metabolic markers
after four weeks consumption of 50 g daily of one of three different
dietary fats, extra virgin coconut oil, butter or extra virgin olive oil, in
healthy men and women in the general population. This was
randomized clinical trial conducted over June and July 2017 in
general community in Cambridgeshire, UK. Volunteer adults were
recruited by the British Broadcasting Corporation through their
websites. Eligibility criteria were men and women aged 50-75 years,
with no known history of cancer, cardiovascular disease or diabetes,
not on lipid lowering medication, no contraindications to a high-fat
diet and willingness to be randomized to consume one of the three
dietary fats for 4 weeks. Of 160 individuals initially expressing an
interest and assessed for eligibility, 96 were randomized to one of
three interventions; 2 individuals subsequently withdrew and 94 men
and women attended a baseline assessment. Their mean age was 60
years, 67% were women and 98% were European Caucasian. Of
these, 91 men and women attended a follow-up assessment 4 weeks
later. Participants were randomized to extra virgin coconut oil, extra
virgin olive oil or unsalted butter and asked to consume 50 g daily of
one of these fats for 4 weeks, which they could incorporate into their
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usual diet or consume as a supplement. The primary outcome was

change in serum LDL-C; secondary outcomes were change in TC and
HDL-C, TC/HDL-C ratio and non-HDL-C; change in weight, body mass
index (BMI), waist circumference, per cent body fat, systolic and
diastolic blood pressure, fasting plasma glucose and C reactive
protein. LDL-C concentrations were significantly increased on butter
compared with coconut oil (+0.42, 95% CI 0.19-0.65 mmol/L,
p<0.0001) and with olive oil (+0.38, 95% CI 0.16-0.60 mmol/L,
p<0.0001), with no differences in change of LDL-C in coconut oil
compared with olive oil (-0.04, 95% CI -0.27-0.19 mmol/L, p=0.74).
Coconut oil significantly increased HDL-C compared with butter
(+0.18, 95% CI 0.06-0.30 mmol/L) or olive oil (+0.16, 95% CI 0.03-
0.28 mmol/L). Butter significantly increased TC/HDL-C ratio and non-
HDL-C compared with coconut oil but coconut oil did not significantly
differ from olive oil for TC/HDL-C and non-HDL-C. There were no
significant differences in changes in weight, BMI, central adiposity,
fasting blood glucose, systolic or diastolic blood pressure among any
of the three intervention groups. Two different dietary fats (butter
and coconut oil) which are predominantly saturated fats, appear to
have different effects on blood lipids compared with olive oil, a
predominantly monounsaturated fat with coconut oil more
comparable to olive oil with respect to LDL-C. The effects of different
dietary fats on lipid profiles, metabolic markers and health outcomes
may vary not just according to the general classification of their main
component fatty acids as saturated or unsaturated but possibly
according to different profiles in individual fatty acids, processing
methods as well as the foods in which they are consumed or dietary
patterns. These findings do not alter current dietary
recommendations to reduce saturated fat intake in general but
highlight the need for further elucidation of the more nuanced
relationships between different dietary fats and health (6).
Brassard & al. (7) revealed that recent evidence suggests that the
association between dietary Saturated Fatty Acids (SFAs) and CAD
risk varies according to food sources. How SFAs from butter and
cheese influence HDL-mediated Cholesterol Efflux Capacity (CEC), a
key process in reverse cholesterol transport, is currently unknown. In
a predefined secondary analysis of a previously published trial, this
study has examined how diets rich in SFAs from either cheese or
butter influence HDL-mediated CEC, compared with diets rich in
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either MonoUnsaturated Fatty Acids (MUFAs) or PolyUnsaturated

Fatty Acids (PUFAs). In a randomized crossover controlled
consumption trial, 46 men and women with abdominal obesity
consumed 5 isocaloric diets, each for 4 weeks. Two diets were rich in
SFAs either from cheese (CHEESE) or butter (BUTTER) [12.4-12.6% of
energy (%E) as SFAs, 32%E as fat, 52%E as carbohydrates]. In 2 other
diets, SFAs (5.8%E) were replaced with either MUFAs from refined
olive oil (MUFA) or PUFAs from corn oil (PUFA). Finally, a lower fat
and carbohydrate diet was used as a control (5.8%E as SFAs, 25.0%E
as fat, 59%E as carbohydrates; CHO). Post-diet HDL-mediated CEC
was determined ex vivo using radiolabelled J774 macrophages
incubated with apolipoprotein B-depleted serum from the
participants. Mean (±SD) age was 41.4 ± 14.2 years, and waist
circumference was 107.6± 11.5 cm in men and 94.3 ± 12.4 cm in
women. BUTTER and MUFA increased HDL-mediated CEC compared
with CHEESE (+4.3%, p=0.026 and +4.7%, p=0.031, respectively).
Exploring the significant diet × sex interaction (p=0.044) revealed that
the increase in HDL-mediated CEC after BUTTER compared with
CHEESE was significant among men (+6.0%, p=0.047) but not women
(+2.9%, p=0.19), whereas the increase after MUFA compared with
CHEESE was significant among women (+9.1%, p=0.008) but not men
(-0.6%, p=0.99). These results provide evidence of a food matrix
effect modulating the impact of dairy SFAs on HDL-mediated CEC
with potential sex-related differences that deserve further
investigation (7).
Tonstad & al. (8) mentioned that margarine leads to lower TC and
LDL-C levels than butter but may contain trans fatty acids that
increase atherogenic lipids. A food company has used data
concerning the cholesterolemic effects of individual fatty acids,
including trans fatty acids, to develop a commercially available and
virtually trans-free margarine. The effect of this novel margarine on
serum lipids and lipoproteins was compared with that of butter in
free-living, hypercholesterolemic subjects. This was a two-period,
outpatient cross-over trial at a university hospital lipid clinic. The
study involved 77 subjects, and was completed by 53 men and 19
women aged 35-65 years with serum TC levels of between 6.0 and
7.9 mmol/L. Two 23-day regimens, separated by a 4-week washout
period, included individualized dietary prescriptions supplemented
with butter or margarine designed to provide 15% of total dietary
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energy. In comparison with butter, margarine intake lowered total

and LDL-C levels by respectively 11.1% (99% CI 8.1-14.1) and 11.3%
(99% CI 7.6-15.1). The reduction in LDL-C was < 3% in nearly one-fifth
of the subjects despite appropriate changes in serum triglyceride
fatty acids. Of the tested clinical and demographic variables, only the
percentage of energy obtained from saturated fat during the
margarine intake period was associated with dietary responsiveness
(explaining 12% of the variation; p<0.01). The results suggest that a
margarine designed to meet nutritional recommendations for
hypercholesterolemia is more efficacious than butter in reducing
atherogenic lipid levels in hypercholesterolemic subjects (8).
de Almeida & al. (9) stated that evidence from in vitro and animal
studies indicates that Conjugated Linoleic Acid (CLA) possesses anti-
diabetic properties, which appear to be attributed to cis-9, trans-11
CLA, the major CLA isomer in ruminant fat. However, there is a
shortage of studies addressing CLA from natural source. The present
study aimed to evaluate the effects of butter naturally enriched in
cis-9, trans-11 CLA on parameters related to glucose tolerance,
insulin sensitivity and dyslipidemia in rats. Forty male Wistar rats
were randomly assigned to the following dietary treatments
(n=10/group), for 60 days: 1) Normal fat-Soybean oil (NF-So): diet
containing 4.0% soybean oil (SO); 2) High Fat-Control Butter (HF-Cb):
diet containing 21.7% control butter and 2.3% SO; 3) High Fat-CLA
enriched Butter (HF-CLAb): diet containing 21.7% cis-9, trans-11 CLA-
enriched butter and 2.3% SO; and 4) High fat-Soybean oil (HF-So):
diet containing 24.0% SO. HF-Cb and HF-CLAb diets contained 0.075%
and 0.235% of cis-9, trans-11 CLA, respectively. HF-CLAb-fed rats had
lower serum insulin levels at fasting than those fed with the HF-Cb
diet, while the PPARγ protein levels in adipose tissue was increased in
HF-CLAb-fed rats compared to HF-Cb-fed rats. R-QUICK was lower in
HF-Cb than in NF-So group, while no differences in R-QUICK were
observed among NF-So, HF-CLAb and HF-So groups. Serum HDL-C
levels were higher in HF-CLAb-fed rats than in those fed NF-So, HF-Cb
and HF-So diets, as well as higher in NF-So-fed rats than in HF-Cb and
HF-So-fed rats. HF-CLAb, HF-Cb and HF-So diets reduced serum LDL-C
levels when compared to NF-So, whereas serum triacylglycerol levels
were increased in HF-CLAb. The data show that feeding rats on a
high-fat diet containing butter naturally enriched in cis-9, trans-11
CLA prevented hyperinsulinemia and increased HDL-C, which could
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be associated with higher levels of cis-9, trans-11 CLA, vaccenic acid,

oleic acid and lower levels of short and medium-chain saturated fatty
acids from butter naturally modified compared to control butter. On
the other hand CLA-enriched butter also increased serum
triacylglycerol levels, which could be associated with concomitant
increases in the content of trans-9 and trans-10 C18:1 isomers in the
CLA-enriched butter (9).
Wood & al. (10) determined the effect of diets containing 50% of
fat calories from butter, butter enriched with mono- and
polyunsaturated fatty acids, and margarines with and without trans
fatty acids on the serum lipids of 38 healthy men in a free-living
condition. Serum lipid responses to the high level of individual
dietary fats were unexpectedly small. The butter diet produced a
small, but significant rise (5%) in the TC and LDL-C, relative to all
other diets. Enrichment of butter with either olive oil (50/50) or
sunflower oil (50/50) failed to reduce serum lipid levels below
habitual diet values. Hard margarine, containing 29% trans fatty
acids, caused a decrease in apolipoprotein A-I and B levels, but did
not change total serum cholesterol or LDL-C levels, relative to
habitual diet values. A soft margarine, high in linoleate, with no trans
fatty acids reduced TC, LDL-C, and apolipoprotein B significantly,
relative to all diets. Soft margarine HDL-C levels remained
unchanged, but apolipoprotein A-I values were decreased relative to
habitual and butter diets. The quantities of saturated fatty acids and
the sum of monounsaturated and polyunsaturated fatty acids
consumed on the hard and soft margarines were equal; therefore,
the different response of serum cholesterol and LDL-C between these
two diets is attributable to the Trans fatty acids in the hard
margarine. The data indicate that Trans fatty acids are not
metabolically equivalent to the natural cis isomers and that they
affect the serum lipid profile adversely (10).

This chapter (1-10) demonstrates that replacing butter by low-

trans soft margarines favorably affects the blood lipoprotein profile
and may reduce the predicted risk of CHD, but high-trans hard
margarines probably confer no benefit over butter.
LDL-C concentrations were significantly increased on butter
consumption.
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The butter diet produced a small, but significant rise (5%) in the
TC and LDL-C, relative to all other diets.
Dietary ghee 10% fed for 4 weeks did not have any significant
effect on levels of serum TC, but did increase triglyceride levels in
Fischer inbred rats.

References
1. Engel S, Tholstrup T. Butter increased total and LDL cholesterol
compared with olive oil but resulted in higher HDL cholesterol compared
with a habitual diet. Am J Clin Nutr. 2015;102(2):309-15.
2. Brassard D, Tessier-Grenier M, Allaire J, et al. Comparison of the
impact of SFAs from cheese and butter on cardiometabolic risk factors: a
randomized controlled trial. Am J Clin Nutr. 2017;105(4):800-9.
3. Zock PL, Katan MB. Butter, margarine and serum lipoproteins.
Atherosclerosis. 1997;131(1):7-16.
4. Denke MA. Effects of cocoa butter on serum lipids in humans:
historical highlights. Am J Clin Nutr. 1994;60(6 Suppl):1014S-6S.
5. Sharma H, Zhang X, Dwivedi C. The effect of ghee (clarified butter)
on serum lipid levels and microsomal lipid peroxidation. Ayu. 2010;
31(2):134-40.
6. Khaw KT, Sharp SJ, Finikarides L, et al. Randomised trial of coconut
oil, olive oil or butter on blood lipids and other cardiovascular risk factors
in healthy men and women. BMJ Open. 2018;8(3):e020167.
7. Brassard D, Arsenault BJ, Boyer M, et al. Saturated fats from butter
but not from cheese increase HDL-mediated cholesterol efflux capacity
from J774 macrophages in men and women with abdominal obesity. J
Nutr. 2018;148(4):573-80.
8. Tonstad S, Strøm EC, Bergei CS, et al. Serum cholesterol response to
replacing butter with a new trans-free margarine in hypercholesterolemic
subjects. Nutr Metab Cardiovasc Dis. 2001;11(5):320-6.
9. de Almeida MM, Luquetti SC, Sabarense CM, et al. Butter naturally
enriched in cis-9, trans-11 CLA prevents hyperinsulinemia and increases
both serum HDL cholesterol and triacylglycerol levels in rats. Lipids Health
Dis. 2014 Dec 22;13:200.
10. Wood R, Kubena K, O'Brien B, et al. Effect of butter, mono- and
polyunsaturated fatty acid-enriched butter, trans fatty acid margarine, and
zero trans fatty acid margarine on serum lipids and lipoproteins in healthy
men. J Lipid Res. 1993;34(1):1-11.
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MORTALITY, CARDIOVASCULAR DISEASE,

GLYCEMIC RESPONSE & DIABETES
van den Brandt & Schouten (1) noticed that nut intake has been
associated with lower mortality, but few studies have investigated
causes of death other than cardiovascular disease (CVD), and dose-
response relationships remain unclear. The relationship of nut (tree
nut, peanut) and peanut butter intake with overall and cause-specific
mortality was investigated. In the Netherlands Cohort Study, 120,852
men and women aged 55-69 years provided information on dietary
and lifestyle habits in 1986. Mortality follow-up until 1996 consisted
of linkage to Statistics Netherlands. Multivariate case-cohort
analyses were based on 8,823 deaths and 3,202 subcohort members
with complete data on nuts and potential confounders. Meta-
analyses of the results with those published from other cohort
studies were conducted. Total nut intake was related to lower
overall and cause-specific mortality (cancer, diabetes, cardiovascular,
respiratory, neurodegenerative diseases, and other causes) in men
and women. When comparing those consuming 0.1-<5, 5-<10 and
10+ g nuts/day with non-consumers, multivariable hazard ratios for
total mortality were 0.88, 0.74 and 0.77 [95% confidence interval (CI),
0.66-0.89+, respectively (p trend = 0.003). Cause-specific hazard
ratios comparing 10+ vs. 0 g/day varied from 0.56 for
neurodegenerative to 0.83 for CVD mortality. Restricted cubic
splines showed nonlinear dose-response relationships with mortality.
Peanuts and tree nuts were inversely related to mortality, whereas
peanut butter was not. In meta-analyses, summary hazard ratios for
highest vs. lowest nut consumption were 0.85 for cancer, and 0.71
for respiratory mortality. The data show that nut intake was related
to lower overall and cause-specific mortality, with evidence for
nonlinear dose-response relationships. Peanut butter was not
related to mortality (1).
Pimpin & al. (2) noticed that dietary guidelines recommend
avoiding foods high in saturated fat. Yet, emerging evidence suggests
cardiometabolic benefits of dairy products and dairy fat. Evidence on
the role of butter, with high saturated dairy fat content, for total
mortality, CVD, and type 2 diabetes remains unclear. The authors
aimed to systematically review and Meta-analyze the association of
butter consumption with all-cause mortality, CVD, and diabetes in
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general populations. Nine databases were searched from inception

to May 2015 without restriction on setting, or language, using
keywords related to butter consumption and cardiometabolic
outcomes. Prospective cohorts or randomized clinical trials providing
estimates of effects of butter intake on mortality, CVD including
coronary hear disease (CHD) and stroke, or diabetes in adult
populations were included. One investigator screened titles and
abstracts; and two reviewed full-text articles independently in
duplicate, and extracted study and participant characteristics,
exposure and outcome definitions and assessment methods, analysis
methods, and adjusted effects and associated uncertainty, all
independently in duplicate. Study quality was evaluated by a
modified Newcastle-Ottawa score. Random and fixed effects meta-
analysis pooled findings, with heterogeneity assessed using the I2
statistic and publication bias by Egger's test and visual inspection of
funnel plots. Nine publications were identified including 15 country-
specific cohorts, together reporting on 636,151 unique participants
with 6.5 million person-years of follow-up and including 28,271 total
deaths, 9,783 cases of incident CVD, and 23,954 cases of incident
diabetes. No RCTs were identified. Butter consumption was weakly
associated with all-cause mortality (n=9 country-specific cohorts; per
14g (1 tablespoon)/day: RR = 1.01, 95% CI 1.00-1.03, p=0.045); was
not significantly associated with any CVD (n=4; RR = 1.00, 95% CI
0.98-1.02; p= 0.704), CHD (n=3; RR = 0.99, 95% CI 0.96-1.03;
p=0.537), or stroke (n=3; RR = 1.01, 95% CI 0.98-1.03; p=0.737), and
was inversely associated with incidence of diabetes (n=11; RR = 0.96,
95% CI 0.93-0.99; p=0.021). Evidence for heterogeneity nor
publication bias was not identified. This systematic review and
meta-analysis suggests relatively small or neutral overall associations
of butter with mortality, CVD, and diabetes. These findings do not
support a need for major emphasis in dietary guidelines on either
increasing or decreasing butter consumption, in comparison to other
better established dietary priorities; while also highlighting the need
for additional investigation of health and metabolic effects of butter
and dairy fat (2).
Liu & al. (3) stated that several recent articles have called into
question the deleterious effects of high animal fat diets due to mixed
results from epidemiologic studies and the lack of clinical trial
evidence in meta-analyses of dietary intervention trials. The authors
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were interested in examining the theoretical effects of substituting

plant-based fats from different types of margarine for animal-based
fat from butter on the risk of atherosclerosis-related CVD. Totally
71,410 women, aged 50-79 years were prospectively studied, and
their risk was evaluated for clinical myocardial infarction (MI), total
CHD, ischemic stroke, and atherosclerosis-related CVD with an
average of 13.2 years of follow-up. Butter and margarine intakes
were obtained at baseline and year 3 by means of a validated food
frequency questionnaire. Cox proportional hazards regression using
a cumulative average diet method was used to estimate the
theoretical effect of substituting 1 teaspoon/day of three types of
margarine for the same amount of butter. Substituting butter or
stick margarine with tub margarine was associated with lower risk of
MI (HRs 0.95 and 0.91). Subgroup analyses, which evaluated these
substitutions among participants with a single source of spreadable
fat, showed stronger associations for MI (HRs 0.92 and 0.87).
Outcomes of total CHD, ischemic stroke, and atherosclerosis-related
CVD showed wide CIs but the same trends as the MI results. This
theoretical dietary substitution analysis suggests that substituting
butter and stick margarine with tub margarine when spreadable fats
are eaten may be associated with reduced risk of MI (3).
Jiang & al. (4) stated that nuts are high in unsaturated
(polyunsaturated and monounsaturated) fat and other nutrients that
may improve glucose and insulin homeostasis. This study examined
prospectively the relationship between nut consumption and risk of
type 2 diabetes. This was a prospective cohort study of 83,818
women from 11 states in the Nurses' Health Study. The women were
aged 34 to 59 years, had no history of diabetes, CVD, or cancer,
completed a validated dietary questionnaire at baseline in 1980, and
were followed up for 16 years. Totally 3,206 new cases of type 2
diabetes were documented. Nut consumption was inversely
associated with risk of type 2 diabetes after adjustment for age, body
mass index (BMI), family history of diabetes, physical activity,
smoking, alcohol use, and total energy intake. The multivariate
Relative risks (RRs) across categories of nut consumption
(never/almost never, <once/week, 1-4 times/week, and ≥5
times/week) for a 28-g (1 oz) serving size were 1.0, 0.92 (95% CI 0.85-
1.00), 0.84 (0.95% CI 0.76-0.93), and 0.73 (95% CI 0.60-0.89) (p for
trend <0.001). Further adjustment for intakes of dietary fats, cereal
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fiber, and other dietary factors did not appreciably change the
results. The inverse association persisted within strata defined by
levels of BMI, smoking, alcohol use, and other diabetes risk factors.
Consumption of peanut butter was also inversely associated with
type 2 diabetes. The multivariate RR was 0.79 (95% CI 0.68-0.91; p
for trend <0.001) in women consuming peanut butter 5 times or
more a week (equivalent to ≥140 g [5 oz] of peanuts/week)
compared with those who never/almost never ate peanut butter.
The findings suggest potential benefits of higher nut and peanut
butter consumption in lowering risk of type 2 diabetes in women. To
avoid increasing caloric intake, regular nut consumption can be
recommended as a replacement for consumption of refined grain
products or red or processed meats (4).
Lilly & al. (5) determine whether supplementation of a high-
glycemic index breakfast meal with peanut butter attenuates the
glycemic response. Sixteen healthy adults, aged 24.1 ± 3.5 years,
reported in the morning to a nutrition assessment laboratory for two
days of data collection, having fasted 8 to 12 hours. On day 1
(control), fasting blood glucose (BG) was measured using
glucometers, then participants consumed two slices of white bread
and 250 mL apple juice (60 g carbohydrate) within 15 minutes. BG
was measured again at 15, 30, 60, 90, and 120 minutes after the first
bite of the meal. On day 2, the protocol was repeated, except 32 g (2
tbsp) of peanut butter was added to the meal (treatment). The spike
in BG was significantly lower on the treatment versus control day
(35.8 ± 16.4 vs. 51.0 ± 20.8 mg/dL, respectively; p<0.01), and BG was
significantly lower on the treatment day at 15, 30, and 60 minutes
post-meal consumption (p<0.05). This study indicates that
supplementation with 32 g (2 tbsp) peanut butter attenuates the
magnitude of BG spike and overall glycemic response to high-
glycemic index meal and may be a practical, beneficial strategy to
prevent undesirable elevations in BG (5).

This chapter (1-5) suggests relatively small or neutral overall

associations of butter of with mortality, CVD, and diabetes.
There is potential benefit of higher nut and peanut butter
consumption in lowering risk of type 2 diabetes in women.
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Substituting butter and stick margarine with tub margarine when

spreadable fats are eaten may be associated with reduced risk of
myocardial infarction.

References
1. van den Brandt PA, Schouten LJ. Relationship of tree nut, peanut
and peanut butter intake with total and cause-specific mortality: a cohort
study and meta-analysis. Int J Epidemiol. 2015;44(3):1038-49.
2. Pimpin L, Wu JH, Haskelberg H , et al. Is butter back? A systematic
review and meta-analysis of butter consumption and risk of cardiovascular
disease, diabetes, and total mortality. PLoS One. 2016;11(6):e0158118.
3. Liu Q, Rossouw JE, Roberts MB, et al. Theoretical effects of
substituting butter with margarine on risk of cardiovascular disease.
Epidemiology. 2017;28(1):145-56.
4. Jiang R, Manson JE, Stampfer MJ, et al. Nut and peanut butter
consumption and risk of type 2 diabetes in women. JAMA. 2002;
288(20):2554-60.
5. Lilly LN, Heiss CJ, Maragoudakis SF, et al. The effect of added peanut
butter on the glycemic response to a high-glycemic index meal: a pilot
study. J Am Coll Nutr. 2018 Nov 5:1-7.

OBESITY
Maioli & al. (1) developed a novel diet based on standard AIN93G
diet that would be able to induce experimental obesity and impair
immune regulation with high concentrations of both carbohydrate
and lipids. This study compared the effects of this high sugar and
butter (HSB) diet with other modified diets, male C57BL/6 mice were
fed either mouse chow, or AIN93G diet, or high sugar (HS) diet, or
high-fat (HF) diet, or high sugar and butter (HSB) diet for 11 weeks ad
libitum. HSB diet induced higher weight gain. Therefore, control
AIN93G and HSB groups were chosen for additional analysis.
Regulatory T cells were studied by flow cytometry, and cytokine
levels were measured by ELISA. Although HF and HSB diets were able
to induce a higher weight gain compatible with obesity in treated
mice, HSB-fed mice presented the higher levels of serum glucose
after fasting and the lowest frequency of regulatory T cells in adipose
tissue. In addition, mice that were fed HSB diet presented higher
levels of cholesterol and triglycerides, hyperleptinemia, increased
resistin and leptin levels as well as reduced adiponectin serum levels.
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It was found increased frequency of CD4(+)CD44(+) effector T cells,

reduction of CD4(+)CD25(+)Foxp3(+) and Th3 regulatory T cells as
well as decreased levels of IL-10 and TGF-β in adipose tissue of HSB-
fed mice. Therefore, HSB represents a novel model of obesity-
inducing diet that was efficient in triggering alterations compatible
with metabolic syndrome as well as impairment in immune
regulatory parameters (1).
Wirths & al. (2) studied the effect of a fat-modified diet with
1,100 kcal (4,600 kJ) on the reduction of body weight and body fat. A
low-carbohydrate, high-fat diet with predominant animal fat (diet T)
was compared with a low-carbohydrate, high-fat diet with
predominant vegetable fat (diet P). Diet T was composed of 52.1% of
the energy as fat, 20.7% as protein and 27.2% as carbohydrates. Diet
P was composed of 54.1% of the energy as fat, 18.9% as protein and
27.0% as carbohydrates. More than two-thirds of the fat in diet T
was butter, in diet P margarine. The study was carried out with 30
subjects (8 men) over two 21-day periods. With diet T, men had a
mean weight loss of 7.1 kg = 338 g/d, of which 3.2 kg = 152 g/d were
proved to be body fat, while women had a mean weight loss of 4.4 kg
= 210 g/d, of which 2.3 kg = 110 g/d were proved to be body fat.
With diet P, men had a mean weight loss of 7,6 kg = 362 g/d, of which
were 3.9 kg = 186 g/d body fat, while women lost 3.8 kg = 181 g/d of
body weight on average, of which were 2.0 kg = 95 g/d body fat. An
improvement of blood pressure was also found. Significant
differences of the reduction of body weight and body fat between
the diet with mostly animal fat and the diet with mostly vegetable fat
were not found (2).

This chapter (1-2) shows that high sugar and butter (HSB) diet
induces obesity and metabolic syndrome with decrease in regulatory
T cells in adipose tissue of mice.
There were insignificant differences of the reduction of body
weight and body fat between the diet with mostly animal fat and the
diet with mostly vegetable fat.

References
1. Maioli TU, Gonçalves JL, Miranda MC et al. High sugar and butter
(HSB) diet induces obesity and metabolic syndrome with decrease in
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regulatory T cells in adipose tissue of mice. Inflamm Res. 2016;65(2):169-

78.
2. Wirths W, Rehage C, Bönnhoff N. Effect of a diet formula with high
butter or margarine content on body weight reduction. Z Ernahrungswiss.
1983;22(3):157-68.

BRONCHIOLITIS
Cavalcanti & al. (1) reported the cases of four patients with
bronchiolitis caused by exposure to artificial butter flavoring at a
cookie factory in Brazil. The clinical, tomographic, and spirometric
findings in the four patients, as well as the lung biopsy findings in one
of the patients were described. All four patients were young male
nonsmokers and developed persistent airflow obstruction (reduced
FEV1/FVC ratio and FEV1 at 25-44% of predicted) after 1-3 years of
exposure to diacetyl, without the use of personal protective
equipment, at a cookie factory. The HRCT findings were indicative of
bronchiolitis. In one patient, the surgical lung biopsy revealed
bronchiolitis obliterans accompanied by giant cells. Bronchiolitis
resulting from exposure to artificial flavoring agents should be
included in the differential diagnosis of airflow obstruction in workers
in Brazil (1).
Egilman & Schilling (2) mentioned that respiratory exposure to
diacetyl and diacetyl-containing flavorings used in butter-flavored
microwave popcorn (BFMP) causes lung disease, including
Bronchiolitis Obliterans (BO), in flavorings and popcorn
manufacturing workers. However, there are no published reports of
lung disease among BFMP consumers. A case series was presented
of three BFMP consumers with biopsy-confirmed BO. Data were
reviewed relating to consumer exposures, estimate case exposures,
and compare them to diacetyl-containing flavoring-exposed
manufacturing workers with lung disease. These consumer cases'
exposure levels are comparable to those that caused disease in
workers. The authors were unable to identify any other exposures or
diseases known or suspected to cause BO in these cases. BFMP
poses a significant respiratory risk to consumers. Some
manufacturers have substituted diacetyl with other alpha-diketones
that are likely to pose a similar risk. Simple consumer practices such
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as cooling the popcorn bag would eliminate the risk of severe lung
disease (2).

This chapter (1-2) shows that artificial butter flavoring are related
to the development of bronchiolitis.

References
1. Cavalcanti Zdo R, Albuquerque Filho AP, Pereira CA, Coletta EN.
Bronchiolitis associated with exposure to artificial butter flavoring in
workers at a cookie factory in Brazil. J Bras Pneumol. 2012;38(3):395-9.
2. Egilman DS, Schilling JH. Bronchiolitis obliterans and consumer
exposure to butter-flavored microwave popcorn: a case series. Int J Occup
Environ Health. 2012;18(1):29-42.

GASTROINTESTINAL
Barros & al. (1) mentioned that mucositis is the most common
side effect due to chemotherapy or radiotherapy. It refers to the
inflammation of intestinal mucous membranes, and it is associated
with complications such as diarrhea, weight loss, and increased
intestinal permeability (IP). This study was designed to evaluate the
effect of diet containing Conjugated linoleic acid (CLA)-enriched
butter on intestinal damage and inflammatory response after 24 h of
5-fluorouracil (5-FU)-induced mucositis. Mice were divided into four
groups: CTL; CLA; 5-FU, and CLA 5-FU, and they were fed for 31 days.
On the 30th experimental day, mucositis was induced by unique
injection of 300 mg/kg of 5-FU. After 24 hours (31st experimental
day), IP was evaluated; ileum and fecal material were collected to
determine cytokine level and MyeloPerOxidase (MPO) activity and
secretory immunoglobulin A (sIgA). The 5-FU group showed an
increase in IP and MPO activity (CTL vs. 5-FU: p<0.05). Additionally,
increased levels of IP and MPO were observed in CLA 5-FU group
compared to those in the test groups (p<0.05). Animals in the CLA 5-
FU group showed reduced concentrations of sIgA (CTL vs. CLA 5-FU:
p<0.05). CLA-enriched butter exacerbating the 5-FU-induced
intestinal damage. Safety concerns regarding the use of CLA require
further investigation (1).
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Butter
Conor Walter

Minckler & al. (2) stated that Jojoba butter is cyanogenic and has
gained attention among herbal supplement consumers due to claims
that it may aid in weight loss. Jojoba butter is extracted from the
seeds of Jojoba shrubs found in the Sonoran Desert. The seeds have
long been recognized as inedible; however clinical symptoms
following ingestion are not well documented. This report describes a
patient who developed restlessness and gastrointestinal complaints
following ingestion of homemade Jojoba seed butter. The patient's
presentation following ingestion is discussed, as well as effective
workup and treatment. In this case, the patient was monitored and
received fluid resuscitation, lorazepam, and diphenhydramine for
symptomatic therapy. This case describes the gastrointestinal
sequela and effective management following ingestion of Jojoba
butter (2).

This chapter (1-2) shows that conjugated linoleic acid-enriched

butter after 24 hours of intestinal mucositis induction exacerbated
the 5-FU-induced intestinal damage.
A case is reported who developed restlessness and
gastrointestinal complaints following ingestion of homemade Jojoba
seed butter.

References
1. Barros PA, Generoso SV, Andrade ME, et al. Effect of conjugated
linoleic acid-enriched butter after 24 hours of intestinal mucositis
induction. Nutr Cancer. 2017;69(1):168-75.
2. Minckler MR, Fisher J, Bowers R, Amini R. Unusual etiology of
gastrointestinal symptoms: the case of jojoba butter. Open Access Emerg
Med. 2017;9:27-9.
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LIVER STEATOSIS
Nalloor & al. (1) noticed that butter is one of the widely used fats
present in the diet. However, there is no satisfactory study available
that evaluates the effect of a high-fat diet containing butter as the
principal fat on the development of Non-alcoholic Fatty Liver Disease
(NAFLD). In the present study, butter was used for the development
of steatosis in Chang liver cells in vitro study and Swiss albino mice in
vivo study. In vitro steatosis was established, and butter was
compared with oleic acid in Chang liver cells using an oil red O (ORO)-
based colorimetric assay. In the in vivo study, a butter-rich special
diet was fed for 15 weeks to mice, which showed no significant
change in body weight. The expression pattern of phosphatase and
tensin homolog (PTEN) and miR-21 was compared by reverse
transcriptase-PCR. Special diet-fed animals showed downregulated
PTEN compared to normal diet-fed animals, while levels of miR-21
remained the same. Elevations in biochemical parameters, viz.,
triglycerides and liver function tests showed symptoms of onset of
NAFLD. Histophathological study of livers of test animals confirmed
mild-to-moderate degree of NAFLD (1).

This chapter (1) shows that long-term exposure to a butter-rich

diet induces mild-to-moderate steatosis in Chang liver cells and Swiss
albino mice models.

Reference
1. Nalloor TJP, Kumar N , Narayanan K, Palanimuthu VR. Long-term
exposure to a butter-rich diet induces mild-to-moderate steatosis in Chang
liver cells and Swiss albino mice models. J Basic Clin Physiol Pharmacol.
2017;28(3):257-65.

ANTI-INFLAMMATORY
Verma & al. (1) mentioned that Shea butter is traditionally used in
Africa for its anti-inflammatory and analgesic effects. In this study
the anti-inflammatory activities of the methanolic extract of shea
butter (SBE) were investigated using lipopolysaccharide (LPS)-induced
murine macrophage cell line J774. It was observed that SBE
significantly reduced the levels of LPS-induced nitric oxide, Tumor
necrosis factor-α (TNF-α), interleukins, 1β (IL-1β), and -12 (IL-12) in
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the culture supernatants in a dose dependent manner. Expression of

pro-inflammatory enzymes, inducible nitric oxide synthase (iNOS),
and cyclooxygenase-2 (COX-2) were also inhibited by SBE. These
anti-inflammatory effects were due to an inhibitory action of SBE on
LPS-induced iNOS, COX-2, TNF-α, IL-1β, and IL-12 mRNA expressions.
Moreover, SBE efficiently suppressed IκB phosphorylation and NF-κB
nuclear translocation induced by LPS. The findings explain the
molecular bases of shea butter's bioactivity against various
inflammatory conditions and substantiate it as a latent source of
novel therapeutic agents (1).
Penedo & al. (2) conducted a conjugated linoleic acid (CLA)
depletion-repletion study to investigate the effects of dietary c9,t11
CLA on C-reactive protein, transcription factor NFκB,
metalloproteinases 2 and 9, inflammatory mediators (adiponectin,
TNFα, IL-2, IL-4, IL-8, IL-10), body composition, and erythrocyte
membrane composition in healthy normal-weight human adults. CLA
depletion was achieved through an 8-week period of restricted dairy
fat intake (depletion phase; CLA intake was 5.2±5.8 mg/day),
followed by an 8-week period in which individuals consumed 20
g/day of butter naturally enriched with c9,t11 CLA (repletion phase;
CLA intake of 1020±167 mg/day). The participants were 29 healthy
adult volunteers (19 women and 10 men, aged 22 to 36 years), with
body mass index between 18.0 and 29.9 kg m(-2). Blood samples
were collected at baseline and at the end of both depletion and
repletion phases. The content of CLA in erythrocytes decreased
during CLA-depletion and increased during CLA-repletion. Intake of
CLA-enriched butter increased the serum levels of anti-inflammatory
IL-10 but reduced transcription factor NFκB in blood and serum levels
of TNFα, IL-2, IL-8 and inactive metalloproteinase-9. Moreover,
reduced activity of metalloproteinases 2 and 9 in serum was
observed during the CLA-repletion period. In contrast, intake of CLA-
enriched butter had no effects on body composition (DXA analysis) as
well as on serum levels of adiponectin, C-reactive protein, and IL-4.
Taken together, the results indicate that the intake of a c9,t11 CLA-
enriched butter by normal-weight subjects induces beneficial
changes in immune modulators associated with sub-clinical
inflammation in overweight individuals (2).
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This chapter (1-2) demonstrates that anti-inflammatory effects of

shea butter is described through inhibition of iNOS, COX-2, and
cytokines via the Nf-κB pathway in LPS-activated J774 macrophage
cells.
Intake of butter naturally enriched with cis9,trans11 conjugated
linoleic acid reduces systemic inflammatory mediators in healthy
young adults.

References
1. Verma N, Chakrabarti R, Das RH, Gautam HK. Anti-inflammatory
effects of shea butter through inhibition of iNOS, COX-2, and cytokines via
the Nf-κB pathway in LPS-activated J774 macrophage cells. J Complement
Integr Med. 2012 Jan 12;9:Article 4.
2. Penedo LA, Nunes JC, Gama MA, et al. Intake of butter naturally
enriched with cis9,trans11 conjugated linoleic acid reduces systemic
inflammatory mediators in healthy young adults. J Nutr Biochem. 2013;
24(12):2144-51.

HYPERSENSITIVITY/ALLERGY
Sausenthaler & al. (1) noticed that it has been hypothesized that
margarine intake is associated with allergic diseases. However, the
epidemiological evidence in children is limited. The aim of the
present study was to assess the relationship between dietary intake
of margarine and butter with eczema and allergic sensitization in 2-
year-old children. Data of 2,582 children at the age of 2 years with
complete information on exposure to diet and allergic outcome were
analyzed in a German prospective birth cohort study (LISA).
Margarine and butter intake were estimated from a semiquantitative
food frequency questionnaire about general fat use at home
combined with questions on the child's spread intake. Multiple
logistic regression analysis was applied comparing predominant
margarine and predominant butter intake with consumption of both
butter and margarine. Predominant margarine intake was positively
associated with lifetime prevalence of symptomatic eczema (aOR
1.71; 95% CI 1.12-2.61) and doctor-diagnosed eczema (aOR 2.10; 95%
CI 1.36-3.25) and allergic sensitization against inhalant allergens (aOR
2.10; 95% CI 1.01-4.41) at the age of 2 years. No statistically
significant associations were found for butter intake. Stratification
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for parental history of atopic diseases indicated that children at high

risk of atopic diseases have higher effect estimates for margarine
intake compared to children without parental history of atopic
diseases. Stratification for sex also showed higher effect estimates in
boys. Children with predominant margarine consumption had an
increased risk for eczema and allergic sensitization, while butter
intake was no predictor for allergic diseases. However, the authors
could not determine whether margarine is a causal risk factor or
whether other lifestyle factors have influenced this association (1).
Calvani & al. (2) noticed that it has been suggested that changes
in dietary habits, particularly increased consumption of omega-6
polyunsaturated fatty acids (PUFA) and decreased consumption of
omega-3 PUFAs may explain the increase in atopic disease seen in
recent years. Furthermore, it seems possible that it is mainly
prenatal or very early life environmental factors that influence the
development of allergic diseases. It has also been suggested that
intrauterine risk factors may act differently if mother themselves
suffer from allergic disease. The aim of this study was to investigate
whether the consumption of fish, butter and margarine during
pregnancy might influence the development of allergic sensitizations
in the offspring. The study population was divided into the offspring
of allergic and non-allergic mothers. This was a retrospective cohort
study enrolling 295 offspring of allergic mothers and 693 of non-
allergic mothers. Information regarding maternal intake of fish,
butter and margarine during pregnancy as well as other prenatal and
perinatal confounding factors were retrospectively assessed by
parental report via a standardized questionnaire. Atopy was
determined by skin-prick tests (SPT) to eight prevalent inhalant
allergens and two foods. In the allergic mothers' group there is no
clear correlation between maternal intakes of fish, butter and
margarine and sensitizations to food or inhalants. In the non-allergic
mothers' group there was no correlation between butter and
margarine intake and food or inhalant sensitizations. On the
contrary, a protective effect of fish intake on SPT positivity was
observed. In particular, frequent maternal intake (2-3 times/wk or
more) of fish reduced the risk of food sensitizations by over a third
(aOR 0.23; 95% CI 0.08-0.69). A similar trend, even if not significant,
was found for inhalants. Finally, even in the whole study population,
i.e. allergic group plus non-allergic group, there was a similar trend
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between increased consumption of fish and decreased prevalence of

SPT positivity for foods. This study shows that frequent intake of fish
during pregnancy may contrast the development of SPT sensitizations
for foods in the offspring of mothers without atopic disease.
Therefore, larger prospective studies are needed, enrolling mothers
with and without allergic disease, to confirm these results (2).
Simonte & al. (3) mentioned that casual skin contact or inhalation
of peanut butter fumes is reported and feared to cause allergic
reactions in highly sensitive children with peanut allergy but has not
been systematically studied. The authors sought to determine the
clinical relevance of exposure to peanut butter by means of
inhalation and skin contact in children with peanut allergy. Children
with significant peanut allergy (recent peanut-specific IgE antibody
concentration >50 kIU/L or evidence of peanut-specific IgE antibody
and one of the following: clinical anaphylaxis, a reported inhalation-
contact reaction, or positive double-blind, placebo-controlled oral
challenge result to peanut) underwent double-blind, placebo-
controlled, randomized exposures to peanut butter by means of
contact with intact skin (0.2 mL pressed flat for 1 minute) and
inhalation (surface area of 6.3 square inches 12 inches from the face
for 10 minutes). Placebo challenges were performed by using soy
butter mixed with histamine (contact), and scent was masked with
soy butter, tuna, and mint (inhalation). Thirty children underwent
the challenges (median age, 7.7 years; median peanut IgE level, >100
kIU/L; 13 with prior history of contact and 11 with inhalation
reactions). None experienced a systemic or respiratory reaction.
Erythema (3 subjects), pruritus without erythema (5 subjects), and
wheal-and-flare reactions (2 subjects) developed only at the site of
skin contact with peanut butter. From this number of participants, it
can be stated with 96% confidence that at least 90% of highly
sensitive children with peanut allergy would not experience a
systemic-respiratory reaction from casual exposure to peanut butter.
The data show that casual exposure to peanut butter is unlikely to
elicit significant allergic reactions. The results cannot be generalized
to larger exposures or to contact with peanut in other forms (flour
and roasted peanuts) (3).
Lavine & Ben-Shoshan (4) hypothesized that household exposure
to allergenic proteins via an impaired skin barrier, such as atopic
dermatitis, may contribute to the development of IgE sensitization.
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Household presence of peanut is a risk factor for the development of

peanut allergy in children. Sunflower seed butter is a peanut-free
alternative to peanut butter, and sunflower seed allergy is an
uncommon but reported entity. A 3 years old boy presented with
oral discomfort that developed almost immediately after he ate
sunflower seeds for the first time. He was given a dose of
diphenhydramine. Subsequently he vomited, and his symptoms
gradually resolved. A similar episode occurred to a commercial snack
made with sunflower seed butter. Skin prick testing demonstrated a
large positive (10 mm wheal) wheal-and-flare response to slurry of
fresh sunflower seed within 3-4 minutes associated with severe
pruritus. This child has an older sibling with confirmed peanut allergy
(PNA). After the PNA diagnosis was made, the family home became
peanut-free. In lieu of peanut butter, sunflower butter was
purchased and eaten frequently by family members, but not by the
child reported herein. Subsequent to the episodes above, the child
ate a bread roll with visible poppy seeds and developed itchy throat,
dyspnea, and urticaria. Epicutaneous skin testing elicited a >10 mm
wheal size within 3-4 minutes in response to a slurry of whole poppy
seeds and 8 mm to fresh pumpkin seed, which had never been
consumed. The data demonstrate that a case of sunflower allergy in
the context of household consumption of sunflower butter has not
yet been reported. It has been suggested that homes which are
intentionally peanut-safe may provide an environment whereby
infants with impaired skin barrier are at increased risk of allergy to
alternative "butter" products being used, via cutaneous exposure to
these products preceding oral introduction to the child (4).
Yanagida & al. (5) performed an oral food challenge (OFC) with 10
g of butter (equivalent of 2.9 mL cow's milk) and 25-mL heated cow's
milk for 68 children with cow's milk-allergy. Thirty-eight children
reacted only to heated cow's milk. Twenty-four children reacted to
neither heated milk nor butter. Thirty-eight (86.4%) of 44 patients
with positive results to the OFC for heated milk could safely tolerate
butter. It is highly likely that even children with cow's milk-allergy
who show positive results to an OFC for heated milk can consume
butter. The milk-specific IgE value indicative of a negative predictive
value of over 95% was 17.8 kUA/L, and patients with low milk-specific
IgE values may be able to safely consume butter. Including butter in
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the diets of patients with milk-allergy after a butter challenge may

improve quality of life (5).
Mathias (6) reported a patient with multiple atopic allergies,
atopic facial dermatitis, and a generalized atopic skin diathesis who
developed 1] angioedema of the lips and tongue following ingestion
of peanut butter, and 2] localized urticarial reactions following direct
skin contact. Open testing with peanut butter demonstrated
probable immunologic contact urticaria (6).
Anderson & al (7) mentioned that concern has been raised over
the association of diacetyl with lung disease clinically resembling
bronchiolitis obliterans in food manufacturing workers. This has
resulted in the need for identification of alternative chemicals to be
used in the manufacturing process. Structurally similar chemicals,
2,3-pentanedione, 2,3-hexanedione, 3,4-hexanedione and 2,3-
heptanedione, used as constituents of synthetic flavoring agents
have been suggested as potential alternatives for diacetyl, however,
immunotoxicity data on these chemicals are limited. The present
study evaluated the dermal irritation and sensitization potential of
diacetyl alternatives using a murine model. None of the chemicals
was identified as dermal irritants when tested at concentrations up
to 50%. Similar to diacetyl (EC3=17.9%), concentration-dependent
increases in lymphocyte proliferation were observed following
exposure to all four chemicals, with calculated EC3 values of 15.4%
(2,3-pentanedione), 18.2% (2,3-hexanedione), 15.5% (3,4-
hexanedione) and 14.1% (2,3-heptanedione). No biologically
significant elevations in local or total serum IgE were identified after
exposure to 25-50% concentrations of these chemicals. These results
demonstrate the potential for development of hypersensitivity
responses to these proposed alternative butter flavorings and raise
concern about the use of structurally similar replacement chemicals.
Additionally, a contaminant with strong sensitization potential was
found in varying concentrations in diacetyl obtained from different
producers (7).

This chapter (1-7) shows that a 3 year old boy presented with oral
discomfort that developed after he ate sunflower seeds and
sunflower seed butter. Here allergy to sunflower seed and sunflower
butter are proposed vehicle for sensitization.
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Fortunately, casual exposure to peanut butter is unlikely to elicit

significant allergic reactions.
Children with cow's milk-allergy who show positive results to an
OFC for heated milk can consume butter.
There is the potential for development of hypersensitivity
responses to some alternative butter flavorings.

References
1. Sausenthaler S, Kompauer I, Borte M, et al. Margarine and butter
consumption, eczema and allergic sensitization in children. The LISA birth
cohort study. Pediatr Allergy Immunol. 2006;17(2):85-93.
2. Calvani M, Alessandri C, Sopo SM, et al.; Lazio Association of
Pediatric Allergology (APAL) Study Group. Consumption of fish, butter and
margarine during pregnancy and development of allergic sensitizations in
the offspring: role of maternal atopy. Pediatr Allergy Immunol. 2006;
17(2):94-102.
3. Simonte SJ, Ma S, Mofidi S, Sicherer SH. Relevance of casual contact
with peanut butter in children with peanut allergy. J Allergy Clin Immunol.
2003;112(1):180-2.
4. Lavine E, Ben-Shoshan M. Allergy to sunflower seed and sunflower
butter as proposed vehicle for sensitization. Allergy Asthma Clin Immunol.
2015;11(1):2.
5. Yanagida N, Minoura T, Kitaoka S. Butter tolerance in children
allergic to cow's milk. Allergy Asthma Immunol Res. 2015;7(2):186-9.
Erratum: [Allergy Asthma Immunol Res. 2016].
6. Mathias CG. Contact urticaria from peanut butter. Contact
Dermatitis. 1983;9(1):66-8.
7. Anderson SE, Franko J, Wells JR, et al. Evaluation of the
hypersensitivity potential of alternative butter flavorings. Food Chem
Toxicol. 2013;62:373-81.

DERMATOLOGICAL
Shea butter applications and benefits include the following (1):
 Excellent daily skin moisturizer for face and body
 Dramatically helps prevent and reduce stretch marks during
pregnancy and weight gain and loss
 Restores elasticity to skin and reduces blemishes and
wrinkles
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 Anti-aging and anti-free radical agent known to increase

circulation to the skin
 Helps reduces acne and scarring
 Excellent moisturizer for both eczema and psoriasis
 Helps soothe skin and diaper rash
 After sun conditioning
 Helps softens tough skin on hands and feet
 Dry, itchy skin and scalp relief
 Use after shaving to reduce razor irritation and bumps
 Provides healing benefits to skin wounds and cracks
 Restores luster to hair

Cope & al. (2) mentioned that dietary fats modulate a wide
variety of T cell functions in mice and humans. This study examined
the effects of four different dietary fats, predominantly
polyunsaturated sunflower oil, margarine, and predominantly
saturated butter, clarified butter, on the T cell-mediated, systemic
suppression of contact hypersensitivity by ultraviolet radiation in the
Skh:HR-1 hairless mouse. Diets containing either 200 g/kg or 50 g/kg
butter or clarified butter as the sole fat source protected against
systemic photoimmunosuppression, whether the radiation source
was unfiltered ultraviolet B (280-320 nm) or filtered solar simulated
ultraviolet radiation (290-400 nm), in comparison with diets
containing either 200 or 50 g/kg margarine or sunflower oil. There
was a linear relationship (r > 0.9) between protection against
photoimmunosuppression and the proportion of clarified butter in
mice fed a series of 200 g/kg mixed fat diets that provided varying
proportions of clarified butter and sunflower oil. The dietary fats did
not modulate the contact hypersensitivity reaction in unirradiated
animals. The observed phenomena were not primary due to the
carotene, tocopherol, cholecalciferol, retinol, lipid hydroperoxide or
the nonfat solid content of the dietary fats used and appeared to be a
result of the different fatty acid composition of the fats (2).
Warocquier-Clerout & al. (3) reported that non-saponifiable lipid
fraction (ICSB) extracted from cocoa shell butter was solubilized in
Dimethylformamide (DMF) and analysed for its biological activity on
growth of rat and human fibroblasts. Non-saponifiables (10 mug ml(-
1)) partially protected cells from toxicity of DMF (1%) and allowed the
growth of fibroblasts cultivated in optimal conditions (10% fetal calf
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serum-FCS, 37 degrees C) or improved the survival of cells

maintained in altered conditions (2.5% FCS, 35 degrees C). At higher
concentration (ICSB 50 mug ml(-1), DMF 1%), the protective effect
was suppressed. ICSB was fractionated by chromatography into four
compounds: sterols, terpenic alcohols, tocopherols and hydrocarbons
+/- carotenoids. This study found that biological activity of ICSB was
mostly due to the major fraction containing sterols (1-3).

This chapter (1-3) shows that butter has many positive effects on
human skin.

References
1. Why shea butter? Available at outofafricashea.com/the-benefits-of-
shea-butter/.
2. Cope RB, Bosnic M, Boehm-Wilcox C, et al. Dietary butter protects
against ultraviolet radiation-induced suppression of contact
hypersensitivity in Skh:HR-1 hairless mice. J Nutr. 1996;126(3):681-92.
3. Warocquier-Clerout R, Sigot M, Ouraghi M, Chaveron H. Non-
saponifiable fraction of cocoa shell butter: effect on rat and human skin
fibroblasts. Int J Cosmet Sci. 1992;14(1):39-46.

NEUROLOGICAL
Karandikar & al. (1) mentioned that the traditional texts
designate Cow Ghee as Medhya Rasayana, beneficial for mental
alertness and memory. There has been concern about increased risk
of cardiovascular disease due to its high percentage of saturated fatty
acids in ghee. Amongst all edible fats, nutrition composition of cow
ghee and butter is comparatively similar. Hence a study was planned
to assess effect of cow ghee on memory and lipid profile. So the aim
of this study is to assess the effect of cow ghee on memory and lipid
profile. Nootropic activity of test drugs was assessed by Elevated
Plus Maze (EPM) and Morris Water Maze (MWM) model. Rats were
divided into four groups namely control, Piracetam, cow ghee and
butter. All drugs were given orally for 21 days. Transfer latency was
measured in EPM model and probe test was done in MWM model.
Cow ghee and butter group showed no significant effect on memory
in EPM and MWM model. There was reduction in weight of animals
in Cow Ghee group and increase in weight with Butter. In both the
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models there was a significant increase in Triglyceride (TG) and Very

Low Density Lipoproteins (VLDL) levels of rats in butter groups and
increase in TG and VLDL of rats with cow ghee in EPM model. The
result of experiment suggests that no beneficial effect cow ghee and
butter on cognition was seen. However, ghee is relatively safer when
compared to Butter in considering lipid profile (1).
Gama & al. (2) noticed that reduced phospholipase A2 (PLA2)
activity has been reported in blood cells and in postmortem brains of
patients with Alzheimer disease (AD), and there is evidence that
Conjugated linoleic acid (CLA) modulates the activity of PLA2 groups
in non-brain tissues. As CLA isomers were shown to be actively
incorporated and metabolized in the brains of rats, the authors
hypothesized that feeding a diet naturally enriched in CLA would
affect the activity and expression of Pla 2 -encoding genes in rat brain
tissue, with possible implications for memory. To test this
hypothesis, Wistar rats were trained for the inhibitory avoidance task
and fed a commercial diet (control) or experimental diets containing
either low CLA- or CLA-enriched butter for 4 weeks. After this period,
the rats were tested for memory retrieval and killed for tissue
collection. Hippocampal expression of 19 Pla 2 genes was evaluated
by qPCR, and activities of PLA2 groups (cPLA2, iPLA2, and sPLA2)
were determined by radioenzymatic assay. Rats fed the high CLA diet
had increased hippocampal mRNA levels for specific PLA2 isoforms
(iPla 2 g6γ; cPla 2 g4a, sPla 2 g3, sPla 2 g1b, and sPla 2 g12a) and
higher enzymatic activity of all PLA2 groups as compared to those fed
the control and the low CLA diet. The increment in PLA2 activities
correlated significantly with memory enhancement, as assessed by
increased latency in the step-down inhibitory avoidance task after 4
weeks of treatment (rs = 0.69 for iPLA2, p<0.001; rs = 0.81 for cPLA2,
p<0.001; and rs = 0.69 for sPLA2, p<0.001). In face of the previous
reports showing reduced PLA2 activity in AD brains, the present
findings suggest that dairy products enriched in cis-9, trans-11 CLA
may be useful in the treatment of this disease (2).
Granic & al. (3) mentioned that healthy dietary patterns (DPs)
have been linked to better cognition and reduced risk of dementia in
older adults, but their role in cognitive functioning and decline in the
very old (aged ≥85 years) is unknown. This study investigated the
association between previously established DPs from the Newcastle
85+ Study and global and attention-specific cognition over 5 years.
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The authors followed up with 302 men and 489 women (1921 birth
cohort from Northeast United Kingdom) for change in global
cognition [measured by the Standardized Mini-Mental State
Examination (SMMSE)] over 5 years and attention (assessed by the
cognitive drug research attention battery) over 3 years. Two-step
clustering was used to derive DPs and mixed models to determine
the relation between DPs and cognition in the presence of the
dementia susceptibility gene. Previously, 3 DPs that differed in
intake of red meat, potato, gravy, and butter and varied with key
health measures were characterized. When compared with
participants in DP1 (high red meat) and DP3 (high butter),
participants in DP2 (low meat) had higher SMMSE scores at baseline
(p<0.001) and follow-ups, and better initial attention (p<0.05).
Membership in DP1 and DP3 was associated with overall worse
SMMSE scores (β = 0.09, p=0.01 and β = 0.08, p=0.02, respectively)
than membership in DP2 after adjustment for sociodemographic
factors, lifestyle, multimorbidity, and body mass index (BMI).
Additional adjustment for apolipoprotein (apoE) ε4 genotype
attenuated the association to nonsignificant in women but not in
men in DP1 (β = 0.13, p=0.02). Participants in DP1 and DP3 also had
overall worse concentration (β = 0.04, p=0.002 and β = 0.028, p=0.03,
respectively) and focused attention (β = 0.02, p=0.01 and β = 0.02,
p=0.03, respectively), irrespective of apoE ε4 genotype, but similar
rate of decline in all cognitive measures over time. The data show
that DPs high in red meat, potato, gravy (DP1), or butter (DP3) were
associated with poor cognition but not with the rate of cognitive
decline in very old adults (3).

This chapter (1-3) shows no beneficial effect cow ghee and butter
on cognition. While conjugated linoleic acid-enriched butter
improved memory and up-regulated phospholipase A2 encoding-
genes in rat brain tissues.

References
1. Karandikar YS, Bansude AS, Angadi EA. Comparison between the
effect of cow ghee and butter on memory and lipid profile of Wistar rats. J
Clin Diagn Res. 2016;10(9):FF11-FF15.
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2. Gama MA, Raposo NR, Mury FB, et al. Conjugated linoleic acid-
enriched butter improved memory and up-regulated phospholipase A2
encoding-genes in rat brain tissue. J Neural Transm (Vienna). 2015;122
(10):1371-80.
3. Granic A, Davies K, Adamson A, et al. Dietary patterns high in red
meat, potato, gravy, and butter are associated with poor cognitive
functioning but not with rate of cognitive decline in very old adults. J Nutr.
2016;146(2):265-74.

PHARMACOKINETICS OF PHENYTOIN
& CARBAMAZEPINE
Sidhu & al. (1) evaluate the effect of butter on the
pharmacokinetics of phenytoin and carbamazepine. In a crossover
study, phenytoin 30 mg/kg and carbamazepine 56 mg/kg were given
orally to New Zealand white rabbits (n=8 for each drug). Blood
samples were drawn at different time intervals from 0-24 hours from
the marginal ear vein after drug administration. After a washout
period of 7 days, butter (5 mg/kg) was administered for 7 days to the
animals. On the 8th day, butter and phenytoin or carbamazepine
were administered simultaneously and the blood samples were
withdrawn at the same time points. Plasma was separated and
stored at -20 degrees C until assayed for phenytoin and
carbamazepine by HPLC and different pharmacokinetic parameters
were calculated. Butter increased the absorption of both phenytoin
and carbamazepine, as there was a significant increase in the Cmax
and AUC(0-alpha) of both drugs after butter administration. No
significant difference in Tmax was observed. In this study, a high fat
diet increases the bioavailability of phenytoin and carbamazepine in
New Zealand white rabbits (1).

Reference
1. Sidhu S, Malhotra S, Garg SK. Influence of high fat diet (butter) on
pharmacokinetics of phenytoin and carbamazepine. Methods Find Exp
Clin Pharmacol. 2004;26(8):634-8.
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CANCER
Rani & Kansal (1) previously showed that cow ghee relative to
soybean oil had a protective effect against carcinogen induced
mammary cancer in rats. The objective of this study was to elucidate
its biochemical mechanism. Two groups of 21 day old rats (20 each)
were fed for 44 wk diet containing cow ghee or soybean oil (10%).
Five animals from each group were sacrificed at 0 day and at 5, 21
and 44 wk for analysis of phase I and phase II pathways enzymes of
carcinogen metabolism. Dietary cow ghee relative to soybean oil
decreased the activities of cytochrome P450 (CYP) enzymes, CYP1A1,
CYP1A2, CYP1B1 and CYP2B1, responsible for activation of carcinogen
in liver. Carcinogen detoxification activities of uridinediphospho-
glucuronosyl transferase (UDPGT) and quinone reductase (QR) in
liver, and γ-glutamyltranspeptidase (GGTP) and QR in mammary
tissue were significantly higher in cow ghee fed rats than in soybean
oil fed rats. The hepatic GGTP activity decreased on soybean oil diet;
while in cow ghee group it remained unaffected. The findings show
that dietary cow ghee compared to soybean oil down regulates the
enzyme activities responsible for carcinogen activation in liver and
upregulates carcinogen detoxification activities in liver and mammary
tissues (1).
More & al. (2) stated that Diacetyl (DA), a natural butter flavorant,
is a causative agent for the lung disease obliterative bronchiolitis.
Mutagenic properties of 1,2-dicarbonyls have previously been
empirically linked to their possible interaction with DNA nucleobases.
This study for the first time identifies chemically the adduct of DA
with 2-deoxyguanosine. Selective reactivity of DA with 5'-
TTTGTTTTT-3' over 5'-TTTTTTTTT-3' indicated its propensity to modify
specifically the guanosine residue. Treatment of plasmid DNA,
pBR322, with DA induced changes in electrophoretic mobility that are
typical of ternary structure disruption. Such DNA nucleobase
interaction of DA translated into increased apoptosis in DA-treated
SH-SY5Y cells in a dose-dependent manner (IC(50) = 0.114 ± 0.0421
mM). The traditional carbonyl scavengers metformin, 2-
thiobarbituric acid, and d-penicillamine protected cells from DA
toxicity in proportion to their rates of reaction with DA, with d-
penicillamine causing a maximal increase in the IC(50) to 5.23 ±
0.0992 mM when co-incubated with DA (2).
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Hashemian & al. (3) mentioned that nut consumption has been
associated with decreased risk of colorectal, endometrial, lung, and
pancreatic cancers. Polyphenols, fiber, vitamins, and minerals in nuts
may confer this observed protective effect. To the authors'
knowledge, no prospective study has evaluated the effect of nut
consumption on esophageal and gastric cancers. The objective was
to evaluate the associations between nut and peanut butter
consumption and the risk of esophageal and gastric cancers and their
different subtypes. In this study, data were used from the NIH-AARP
Diet and Health Study, which enrolled 566,407 persons who were 50-
71 yr old at baseline (1995-1996). The median follow-up time was
15.5 yr. Intakes of nuts and peanut butter were assessed through the
use of a validated food-frequency questionnaire. Cox proportional
hazard models were used to estimate HRs and 95% CIs for
esophageal and gastric cancers and their subtypes. Totally, identified
966 incident cases of esophageal adenocarcinomas, 323 cases of
esophageal squamous cell carcinoma, 698 cases of gastric cardia
adenocarcinoma, and 732 cases of gastric noncardia adenocarcinoma
were identified. Compared with those who did not consume nuts or
peanut butter [lowest category of consumption (C0)], participants in
the highest category of nut consumption (C3) had a lower risk of
developing gastric noncardia adenocarcinoma [C3 compared with C0,
HR 0.73 (95% CI 0.57-0.94)]. This inverse association was also seen
for peanut butter consumption [C3 compared with C0, HR 0.75 (95%
CI 0.60-0.94)]. No significant associations were observed between
the highest and lowest intakes of nuts or peanut butter and the risk
of gastric cardia adenocarcinoma, esophageal adenocarcinoma, or
esophageal squamous cell carcinoma. The data show that among
older American adults, both nut and peanut butter consumption
were inversely associated with the risk of gastric noncardia
adenocarcinoma (3).
Nieuwenhuis & van den Brandt PA (4) mentioned that nut
consumption has been associated with reduced cancer-related
mortality. However, it is unclear whether nut consumption also
reduces the risk of esophageal and gastric cancer subtypes. This
study prospectively investigated the relationship of tree nut, peanut,
and peanut butter intake with risk of esophageal squamous cell
carcinoma (ESCC), esophageal adenocarcinoma (EAC), gastric cardia
adenocarcinoma (GCA), and gastric non-cardia adenocarcinoma
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(GNCA) in the Netherlands Cohort Study. In 1986, 120,852 males and

females, aged 55-69 years, completed a baseline questionnaire on
diet and cancer risk factors. After 20.3 years of follow-up, 133 ESCC,
200 EAC, 191 GCA, and 586 GNCA cases, and 3,720 subcohort
members were available for multivariable Cox regression analyses,
using a case-cohort approach. Increased total nut consumption was
significantly associated with a decreased risk of ESCC and GNCA [HRs
(95% CIs) for 10 + g/day vs. nonconsumers = 0.54 (0.30-0.96) and 0.73
(0.55-0.97), respectively], but not with EAC and GCA risk. Similar
trends were observed for tree nut and peanut intake, which were
mostly nonsignificant. For peanut butter intake, no significant
associations were found. When excluding the first four years of
follow-up to reduce the possible influence of reversed causation, the
relation between nut consumption and ESCC risk attenuated, but
remained inverse. The findings suggest that increased tree nut and
peanut consumption is inversely associated with GNCA risk and
possibly with ESCC risk, but not with the risk of the other esophageal
and gastric cancer subtypes (4).
van den Brandt & Nieuwenhuis (5) noticed that nut intake has
been associated with reduced mortality and risk of cardiovascular
diseases, but there is only limited evidence on cancer. The
relationship between nut intake and risk of postmenopausal breast
cancer, and estrogen/progesterone receptor (ER/PR) subtypes was
investigated. In The Netherlands Cohort Study, 62,573 women aged
55-69 years provided information on dietary and lifestyle habits in
1986. After 20.3 years of follow-up, 2,321 incident breast cancer
cases and 1,665 subcohort members were eligible for multivariate
case-cohort analyses. Total nut intake was significantly inversely
related to ER negative (ER -) breast cancer risk, with HR 0.55 (95% CI
0.33-0.93) for those consuming at least 10 g nuts/day versus non-
consumers (p trend = 0.025). There were no significant inverse
associations with ER + or total breast cancer. While there was no
variation between PR subtypes, the ER-PR- subtype was also
significantly inversely associated with nut intake, with HR 0.53 (95%
CI 0.29-0.99), p trend = 0.037. Intake of peanuts and tree nuts
separately was also inversely related to ER - breast cancer subtypes,
while no associations were found with peanut butter intake. The
findings suggest an inverse association between nut intake and ER -
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breast cancer, and no association with total or hormone receptor-

positive subtypes (5).
Boudewijns & al. (6) reported that the consumption of nuts has
been associated with a reduction of cancer risk, but only a few
studies have examined the effects of nuts on prostate cancer risk.
The current study prospectively investigated the association between
the consumption of total nuts, tree nuts, peanuts, and peanut butter
and the risk of total, advanced, and non-advanced prostate cancer.
The association between nuts and prostate cancer was evaluated in
the Netherlands Cohort Study, which was conducted among 58,279
men aged 55-69 year at baseline. A case-cohort approach was used
for data processing and analyses. After 20.3 years of follow-up, 3,868
incident prostate cancer cases and 1979 subcohort members were
available for multivariable Cox regression analyses. For total,
advanced, and non-advanced prostate cancer, no significant
associations were found for total nuts (total prostate cancer: hazard
ratio (HR) (95% CI) for 10+ g/day vs. non-consumers = 1.09 (0.92-
1.29), ptrend = 0.409). No significant associations were observed for
tree nuts and peanuts for total, advanced, and non-advanced
prostate cancer risk. Peanut butter consumption was associated with
a significantly increased risk of non-advanced prostate cancer (HR
(95% CI) for 5+ g/day vs. non-consumers = 1.33 (1.08-1.63),
ptrend = 0.008), but not with total or advanced prostate cancer. The
data demonstrate no significant associations were found between
total nut, tree nut, and peanut consumption and total, advanced, and
non-advanced prostate cancer. Peanut butter might be associated
with an increased non-advanced prostate cancer risk (6).
Nieuwenhuis & van den Brandt (7) noticed that nut intake has
been associated with decreased cancer-related mortality, but few
studies have examined the potential of nuts in the chemoprevention
of pancreatic cancer. This study prospectively investigated the
association of total nut, tree nut, peanut, and peanut butter
consumption with pancreatic cancer risk. In the Netherlands Cohort
Study, 120,852 men and women completed a baseline questionnaire,
including a food frequency questionnaire, in 1986. After 20.3 years
of follow-up, 583 incident pancreatic cancer cases, including 349
microscopically confirmed pancreatic cancer (MCPC) cases, were
included in multivariable case-cohort analyses. Increased total nut
consumption was associated with a nonsignificantly decreased MCPC
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risk in men [HR (95% CI) for 10+ g/d vs. nonconsumers = 0.72 (0.47-
1.11), ptrend= 0.163]. No clear association was found in women. For
tree nut and peanut consumption, nonsignificant inverse associations
were observed in men. In women, no or unclear associations were
found for tree nut and peanut consumption. Peanut butter intake
was related to a significantly reduced risk of MCPC in men [HR (95%
CI) for 5+ g/d vs. nonconsumers = 0.53 (0.28-1.00), ptrend= 0.047], but
this relation was not clear in women. Evidence for a nonlinear dose-
response relation with MCPC was found for tree nut intake only. The
associations were weaker when looking at total pancreatic cancer.
The results suggest that nuts and peanut butter might reduce
pancreatic cancer risk in men. In women, no or unclear associations
were found. Nut consumption might reduce the risk of pancreatic
cancer in men (7).
Omer & al. (8) reported that Hepatocellular Carcinoma (HCC) is
one of the major cancers in the world. In Sudan the incidence is
thought to be high and increasing. This study aims to assess the
association between peanut butter intake, as a source of aflatoxins,
and the GSTM1 genotype in the etiology of HCC. A case control study
was conducted among 150 patients and 205 controls from two
regions in Sudan. Food habits with special reference to peanut butter
consumption, as well as peanut storage systems, have been
investigated, as well as confounders such as hepatitis, drinking and
smoking habits, and demographic characteristics. GSTM1 genotype
was assessed in DNA extracted from blood samples (110 cases, 189
controls). A positive association was observed for highest vs. lowest
quartile of peanut butter intake, humid storage system and HCC, with
ORs (95% CI) being 3.0 (1.6-5.5) and 1.6 (1.1-2.5) respectively. The
positive association with peanut butter intake was essentially limited
to subjects with GSTM1 null genotype with OR for highest vs. lowest
quartile 16.7 (2.7-105). The data show that peanut butter
consumption has been identified as a strong risk factor of HCC in a
region with endemic aflatoxin contamination in Sudan and was
essentially limited to subjects with the GSTM1 null genotype (8).
Balasubramanian & al. (9) synthesized solid lipid nanoparticles
carrying a chemotherapeutic payload (i.e., temozolomide, TMZ) were
synthesized using ghee, a clarified butter commonly used in
traditional medicine and food products. Ghee solid lipid
nanoparticles (GSLN) were characterized through dynamic light
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scattering, scanning electron microscopy, and UV-visible

spectrometry. Formulations were generated with varying ratios of
surfactant to lipid, resulting in a maximum TMZ entrapment
efficiency of ˜70%. Optimal formulations were found to have an
average size and polydispersity of ˜220 nm and 0.340, respectively.
Release kinetics revealed TMZ-loaded GSLN (TMZ@GSLN) retained
10% of its pay-load at 2 hours with ˜53% released in 5 hours.
Metabolic activity on human umbilical vein endothelial cells (HUVEC)
revealed GSLN treatment resulted in an increase in viability following
3 days while treatment of glioblastoma LN-229 cells with TMZ@GSLN
resulted in a significant decrease. Evaluation of diffusion of TMZ
across a reconstructed HUVEC monolayer demonstrated TMZ@GSLN
resulted in a significantly higher diffusion of drug when compared to
free TMZ. This data suggests GSLN pose a promising delivery vehicle
for TMZ-based therapeutics. Collectively, this data demonstrates
GSLN exhibit favorable drug carrier properties with anti-proliferative
properties in glioblastoma cancer cells (9).

This chapter (1-9) shows that cow ghee relative to soybean oil
have a protective effect against carcinogen induced mammary cancer
in rats.
The butter flavorant, diacetyl, forms a covalent adduct with 2-
deoxyguanosine, uncoils DNA, and leads to cell death.
Both nut and peanut butter consumption is inversely associated
with the risk of gastric noncardia adenocarcinoma. Increased tree
nut and peanut consumption is inversely associated with GNCA risk
and possibly with ESCC risk, but not with the risk of the other
esophageal and gastric cancer subtypes. For peanut butter intake, no
significant associations are found.
Nuts and peanut butter might reduce pancreatic cancer risk in
men. In women, no or unclear associations are found.

References
1. Rani R, Kansal VK. Effects of cow ghee (clarified butter oil) &
soybean oil on carcinogen-metabolizing enzymes in rats. Indian J Med Res.
2012;136(3):460-5.
2. More SS, Raza A, Vince R. The butter flavorant, diacetyl, forms a
covalent adduct with 2-deoxyguanosine, uncoils DNA, and leads to cell
death. J Agric Food Chem. 2012;60(12):3311-7.
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3. Hashemian M, Murphy G, Etemadi A, et al. Nut and peanut butter

consumption and the risk of esophageal and gastric cancer subtypes. Am J
Clin Nutr. 2017;106(3):858-64.
4. Nieuwenhuis L, van den Brandt PA. Tree nut, peanut, and peanut
butter consumption and the risk of gastric and esophageal cancer
subtypes: the Netherlands Cohort Study. Gastric Cancer. 2018;27(3):274-
84.
5. van den Brandt PA, Nieuwenhuis L. Tree nut, peanut, and peanut
butter intake and risk of postmenopausal breast cancer: The Netherlands
Cohort Study. Cancer Causes Control. 2018;29(1):63-75.
6. Boudewijns EA, Nieuwenhuis L, Geybels MS, et al. Total nut, tree
nut, peanut, and peanutbutter intake and the risk of prostate cancer in the
Netherlands Cohort Study. Prostate Cancer Prostatic Dis. 2019 Jan 28.
[Epub ahead of print]
7. Nieuwenhuis L, van den Brandt PA. Total nut, tree nut, peanut, and
peanut butter consumption and the risk of pancreatic cancer in the
Netherlands Cohort Study. Cancer Epidemiol Biomarkers Prev. 2018;
27(3):274-84.
8. Omer RE, Verhoef L, Van't Veer P, et al. Peanut butter intake,
GSTM1 genotype and hepatocellular carcinoma: a case-control study in
Sudan. Cancer Causes Control. 2001;12(1):23-32.
9. Balasubramanian K, Evangelopoulos M, Brown BS, et al. Ghee
Butter as a Therapeutic Delivery System. J Nanosci Nanotechnol. 2017;
17(2):977-82.

CONTAMINATION
BACTERIAL AGENTS
Dong & al. (1) reported that in 2007, a nationwide Salmonella
Tennessee outbreak occurred via contaminated peanut butter. Here,
a single-nucleotide polymorphism (SNP)-typing method for S.
Tennessee was developed to determine the clonal subtypes of S.
Tennessee that were associated with the peanut butter outbreak.
One seventy-six S. Tennessee isolates from various sources, including
humans, animals, food, and the environment, were analyzed by using
the SNP technique. Eighty-four representative SNP markers were
selected by comparing the sequences of three representative S.
Tennessee strains with different multi-locus sequence typing and
variable number tandem repeats from the collection. The set of
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eighty-four SNP markers showed 100% typeability for the 176 strains,
with the nucleotide diversity ranging from 0.011 to 0.107 (mean =
0.049 ± 0.018, median = 0.044) for each marker. Among the four
clades and nine subtypes generated by the SNP typing, subtype 1,
which comprised 142 S. Tennessee strains, was the most
predominant. The dominance of single-strain clones in subtype 1
revealed that S. Tennessee is highly clonal regardless of outbreak-
association, source, or period of isolation, suggesting the presence of
an S. Tennessee strain prototype. Notably, a minimum 18 SNP set
was able to determine clonal S. Tennessee strains with similar
discrimination power, potentially allowing more rapid and economic
strain genotyping for both outbreaks and sporadic cases. The data
show that the SNP-typing method described here might aid the
investigation of the epidemiology and microevolution of pathogenic
bacteria by discriminating between outbreak-related and sporadic
clinical cases. In addition, this approach enables us to understand
the population structure of the bacterial subtypes involved in the
outbreak (1).

Peanut butter. Potential Salmonella Contamination in Peanut Butter

Leads to Recall

Wilson & al. (2) stated that establishing an association between

possible food sources and clinical isolates requires discriminating the
suspected pathogen from an environmental background, and
distinguishing it from other closely-related foodborne pathogens.
Whole genome sequencing (WGS) to Salmonella subspecies enterica
serotype Tennessee (S. Tennessee) was used to describe genomic
diversity across the serovar as well as among and within outbreak
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clades of strains associated with contaminated peanut butter.

Seventy one isolates of S. Tennessee from disparate food,
environmental, and clinical sources and 2 other closely-related
Salmonella serovars as outgroups (S. Kentucky and S. Cubana), which
were also shot-gun sequenced. A whole genome SNP analysis was
performed using a maximum likelihood approach to infer
phylogenetic relationships. Several monophyletic lineages of S.
Tennessee with limited SNP variability were identified that
recapitulated several food contamination events. S. Tennessee
clades were separated from outgroup salmonellae by more than
sixteen thousand SNPs. Intra-serovar diversity of S. Tennessee was
small compared to the chosen outgroups (1,153 SNPs), suggesting
recent divergence of some S. Tennessee clades. Analysis of all 1,153
SNPs structuring an S. Tennessee peanut butter outbreak cluster
revealed that isolates from several food, plant, and clinical isolates
were very closely related, as they had only a few SNP differences
between them. SNP-based cluster analyses linked specific food
sources to several clinical S. Tennessee strains isolated in separate
contamination events. Environmental and clinical isolates had very
similar whole genome sequences; no markers were found that could
be used to discriminate between these sources. Finally, SNPs were
identified within variable S. Tennessee genes that may be useful
markers for the development of rapid surveillance and typing
methods, potentially aiding in trace back efforts during future
outbreaks. Using WGS can delimit contamination sources for
foodborne illnesses across multiple outbreaks and reveal otherwise
undetected DNA sequence differences essential to the tracing of
bacterial pathogens as they emerge (2).
Shachar & Yaron (3) mentioned that recent large foodborne
outbreaks caused by Salmonella enterica serovars have been
associated with consumption of foods with high fat content and
reduced water activity, even though their ingredients usually
undergo pasteurization. The present study was focused on the heat
tolerance of Salmonella enterica serovars Agona, Enteritidis, and
Typhimurium in peanut butter. The Salmonella serovars in the
peanut butter were resistant to heat, and even at a temperature as
high as 90 degrees C only 3.2-log reduction in CFU was observed. The
obtained thermal inactivation curves were upwardly concave,
indicating rapid death at the beginning (10 min) followed by lower
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death rates and an asymptotic tail. The curves fitted the nonlinear
Weibull model with beta parameters < 1, indicating that the
remaining cells have a lower probability of dying. beta at 70 degrees
C (0.40 +/- 0.04) was significantly lower than beta at 80 degrees C
(0.73 +/- 0.19) and 90 degrees C (0.69 +/- 0.17). Very little decrease
in the viable population (less than 2-log decrease) was noted in
cultures that were exposed to a second thermal treatment. Peanut
butter is a highly concentrated colloidal suspension of lipid and water
in a peanut meal phase. It was hypothesized that differences in the
local environments of the bacteria, with respect to fat content or
water activity, explained the observed distribution and high portion
of surviving cells (0.1%, independent of the initial cell number).
These results demonstrate that thermal treatments are inadequate
to consistently destroy Salmonella in highly contaminated peanut
butter and that the pasteurization process cannot be improved
significantly by longer treatment or higher temperatures (3).
He & al. (4) found significant differences (p<0.05) between the
survival rates of Salmonella enterica and Escherichia coli O157:H7 in
peanut butter with different formulations and water activity. High
carbohydrate content in peanut butter and low incubation
temperature resulted in higher levels of bacterial survival during
storage but lower levels of bacterial resistance to heat treatment (4).
Lathrop & al. (5) stated that peanuts and peanut-based products
have been the source of recent Salmonella outbreaks worldwide.
Because peanut butter is commonly used as an ingredient in baked
goods, such as cookies, the potential risk of Salmonella remaining in
these products after baking needs to be assessed. This research
examines the potential hazard of Salmonella in peanut butter cookies
when it is introduced via the peanut-derived ingredient. The survival
of Salmonella during the baking of peanut butter cookies was
determined. Commercial, creamy-style peanut butter was artificially
inoculated with a five-strain Salmonella cocktail at a target
concentration of 10(8) CFU/g. The inoculated peanut butter was
then used to prepare peanut butter cookie dough following a
standard recipe. Cookies were baked at 350°F (177°C) and were
sampled after 10, 11, 12, 13, 14, and 15 min. Temperature profiles of
the oven and cookies were monitored during baking. The water
activity and pH of the inoculated and uninoculated peanut butter,
raw dough, and baked cookies were measured. Immediately after
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baking, cookies were cooled, and the survival of Salmonella was

determined by direct plating or enrichment. After baking cookies for
10 min, the minimum reduction of Salmonella observed was 4.8 log.
In cookies baked for 13 and 14 min, Salmonella was only detectable
by enrichment reflecting a Salmonella reduction in the range of 5.2 to
6.2 log. Cookies baked for 15 min had no detectable Salmonella.
Results of this study showed that proper baking will reduce
Salmonella in peanut butter cookies by 5 log or more (5).
Burnett & al. (6) reported that in 1996, the first documented
outbreak of salmonellosis associated with the consumption of peanut
butter was reported. This study was undertaken to determine
survival characteristics of high (5.68 log10 cfu g(-1)) and low (1.51
log10 cfu g(-1)) inocula of a five-serotype mixture of Salmonella in
five commercial peanut butters and two commercial peanut butter
spreads. Populations in samples inoculated with 5.68 log10 cfu g(-1)
and stored for 24 weeks at 21 or 5 degrees C decreased 4.14-4.50
log10 cfu g(-1) and 2.86-4.28 log10 cfu g(-1), respectively, depending
on the formulation. The order of retention of viability was: peanut
butter spreads > traditional (regular) and reduced sugar, low-sodium
peanut butters > natural peanut butter. Differences in rates of
inactivation are attributed to variation in product composition as well
as size and stability of water droplets in the colloidal matrix, which
may influence nutrient availability. With the exception of natural
peanut butter, products initially inoculated with 1.51 log10 cfu of
Salmonella g(-1) (32 cfu g(-1)) were positive for the pathogen after
storage for 24 weeks at 5 degrees C. At 21 degrees C, however, with
the exception of one peanut butter spread, all products were
negative for Salmonella after storage for 24 weeks. Post-process
contamination of peanut butter and spreads with Salmonella may to
result in survival in these products for the duration of their shelf life
at 5 degrees C and possibly 21 degrees C, depending on the
formulation (6).
Grasso & al. (7) mentioned that about 1.2 billion pounds of
peanut butter are consumed annually in the U.S. In 2008 to 2009, an
outbreak involving Salmonella Typhimurium in peanut butter led to a
recall of over 3,900 products by over 200 companies. More than 700
people became sick, 100 were hospitalized, and 9 people died from
this outbreak. This study examines the efficacy of high-pressure
processing (HPP) to decrease S. Typhimurium American Type Culture
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Collection (ATCC) 53,647 inoculated into peanut butter and model

systems. The viability of S. Typhimurium in peanut butter stored at
room temperature was investigated. A culture of S. Typhimurium
(6.88 log CFU/g) was inoculated into peanut butter. Following 28 d at
20°C there was a 1.23-log reduction. Approximately 10(6) to 10(7)
CFU/g S. Typhimurium were inoculated into 4 brands of
peanut butter, 3 natural peanut butters and peanut flour slurries at
2, 5, and 10% peanut flour protein in peanut oil and in distilled water.
All were treated at 600 MPa for 5 min at 45°C. While significant
differences were found between natural peanut butter and peanut
protein mixtures, the reduction was <1.0 log. The peanut flour/oil
mixtures had a 1.7, 1.6, and 1.0-log reduction from HPP (2, 5, and
10% protein, respectively) whereas peanut flour/water mixtures had
a 6.7-log reduction for all protein levels. Oil had a protective effect
indicating HPP may not help the microbial safety of water-in-oil food
emulsions including peanut butter. There have been multiple
outbreaks of foodborne illness involving peanut butter products. This
study looks at the potential use of high-pressure processing to reduce
the bacteria that may be in peanut butter (7).
Matak & al. (8) studied the effect of electron beam (e-beam)
radiation on the recovery of Salmonella serotypes Tennessee (ATCC
10722) and Typhimurium (ATCC 14028) in creamy peanut butter over
a 14-d storage period at 22°C was studied. Each Salmonella type was
independently inoculated into peanut butter and subjected to e-
beam doses that ranged from 0 to 3.1 kGy, confirmed by film
dosimetry. After 2-, 4-, 6-, 8-, and 14-d of storage, microbial analyses
were conducted. Survivors were recovered on growth and selective
media using standard spread-plating methods. Microbial counts
(CFU/g) were log-converted and differences were determined by
ANOVA and Tukey's Honestly Significant Differences test. When
samples were not e-beam-treated, there were no significant changes
(p>0.05) in microbial numbers over time. In e-beamed samples,
microbial numbers decreased over time; however, reductions were
not always significant. Initial recovery rates (R-rates) 2 d after e-
beam treatment were significantly different for the 2 strains of
Salmonella and between recovery media (p<0.05); however, these
differences did not persist for the remainder of the storage period
(p>0.05) indicating that injured cells were not able to survive in the
high-fat, low-water activity peanut butter environment. R-rates for
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both strains of Salmonella were maintained until day 14 when there

were significant reductions in Salmonella Typhimurium p<0.05).
These results indicate that Salmonella Tennessee and Salmonella
Typhimurium will survive in peanut butter when exposed to
nonlethal doses of e-beam irradiation. Electron beam (e-beam)
irradiation is an alternative to thermal processing; this technique
inactivates microorganisms and insects that might be present in a
food by generating radiation by accelerated electrons that inactivate
organisms directly because of interaction with cell components and
indirectly by producing free radicals that disrupt integrity of the cell
membrane. E-beam radiation will reduce the number of probable
microbiological hazards that could be present while the food remains
generally unaffected in texture, taste, and nutritional value. A recent
study showed e-beam irradiation to be effective at reducing both
Salmonella Tennessee and Typhimurium in peanut butter by one log
after exposure to less than 1 kGy, highlighting the need to explore
this process further (8).
Ma & al. (9) determined the rates of thermal inactivation of three
Salmonella Tennessee strains in peanut butter associated with an
outbreak and to compare them to the rates of inactivation of
Salmonella strains of other serotypes (Enteritidis, Typhimurium, and
Heidelberg) (SSOS) and of clinical isolates of Salmonella Tennessee
from sporadic cases (STSC). Commercial peanut butter was
inoculated with Salmonella isolates and heated at 71, 77, 83, and 90
degrees C. The thermal inactivation curves were upwardly concave,
indicating rapid death at the beginning (20 min) of heating followed
by lower death rates thereafter. The first-order kinetics approach
and nonlinear Weibull model were used to fit the inactivation curves
and describe the rates of thermal inactivation of Salmonella in
peanut butter. The calculated minimum times needed to obtain a 7-
log reduction at 90 degrees C for the composited three outbreak-
associated strains were significantly greater (p<0.05) than those of
SSOS and STSC. Approximately 120 min were needed to reduce the
outbreak strains of Salmonella Tennessee by 7 log, whereas 86 and
55 min were needed for SSOS and STSC, respectively. These results
indicate that the outbreak-associated Salmonella strains were more
thermotolerant than the other Salmonella strains tested, and this
greater thermal resistance was not serotype specific. Thermal
treatments of peanut butter at 90 degrees C for less than 30 min are
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not sufficient to kill large populations (5 log CFU/g) of Salmonella in

highly contaminated peanut butter (9).
Kottapalli & al. (10) reported that recent recalls and outbreaks
due to foodborne pathogens in thermally processed low-moisture
foods highlight the need for the food industry to validate their
thermal process. The purpose of this study was to validate baking as
an adequate lethality step in controlling Salmonella and Listeria
monocytogenes during the production of peanut butter (PB)-filled
pretzels and whole wheat (WW) pita chips. Two dough types, PB-
filled pretzel and WW pita chip with varying water activities (0.96 to
0.98), were inoculated (target level, ∼108 to 109 CFU/g) with a
multistrain cocktail of Salmonella and L. monocytogenes in separate
trials and were baked at 300°F (148.9°C) and 350°F (176.6°C) for 0, 5,
10, 17, 25, and 30 min. Following baking, samples were rapidly
cooled and analyzed for Salmonella and L. monocytogenes by the
pour plate method. Uninoculated samples were analyzed for total
viable aerobic plate count (APC) and Enterobacteriaceae counts.
Water activity analysis was also performed. The experiment was
replicated three times. Nonlinear regression was used to estimate
the baking times required to achieve a minimum of 4- and 5-log
reduction in APC, Salmonella, and L. monocytogenes. A 4- and 5-log
reduction in APC was predicted following a treatment at 350°F for 3.3
and 5.6 min in WW pita chip product, respectively. Following a
treatment of 350°F for 10 and 25 min, Enterobacteriaceae and APC
counts were below the detection limit (<1 log CFU/g), respectively, in
all of the PB-filled pretzel samples. Salmonella and L. monocytogenes
counts decreased with increasing baking time regardless of the
temperature used. Significant reductions (≥5-log reduction) were
estimated in Salmonella and L. monocytogenes in product baked at
350°F for 15.5 and 17.5 min in WW pita chip dough and PB-filled
pretzel dough, respectively. Both pathogens were below the
detection limit (<1 log CFU/g) in PB-filled pretzel and WW pita chip
products under baking conditions of 350°F for 25 and 30 min,
respectively. This study demonstrates that PB-filled pretzel and WW
pita chip products, when baked to saleable quality, will not present a
public health risk from the standpoint of Salmonella or L.
monocytogenes (10).
Klu & Chen (11) observed the behavior of probiotics and selected
bacterial pathogens co-inoculated into peanut butter during
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gastrointestinal simulation. Peanut butter homogenates co-

inoculated with Salmonella/Listeria strains (5 log CFU ml(-1)) and
lyophilized or cultured probiotics (9 log CFU ml(-1)) were exposed to
simulated gastrointestinal conditions for 24 hours at 37°C. Sample
pH, titratable acidity and pathogen populations were determined.
Agar diffusion assay was performed to assess the inhibitory effect of
probiotic culture supernatants with either natural (3.80
(Lactobacillus), 3.78 (Bifidobacteirum) and 5.17
(Streptococcus/Lactococcus)) or neutralized (6·0) pH. Antibacterial
effect of crude bacteriocin extracts were also evaluated against the
pathogens. After 24 hours, samples with probiotics had lower pH
and higher titratable acidity than those without probiotics. The
presence of probiotics caused a significant reduction (p<0·05) in
pathogen populations. Supernatants of Bifidobacterium and
Lactobacillus cultures inhibited pathogen growth; however, the
elevation of pH diminished their antibacterial activities. Crude
bacteriocin extracts had a strain-specific inhibitory effect only
towards Listeria monocytogenes. Probiotics in 'peanut butter'
survived simulated gastrointestinal conditions and inhibited the
growth of Salmonella/Listeria. Peanut butter is a plausible carrier to
deliver probiotics to improve the gastrointestinal health of children in
developing countries (11).

This chapter (1-11) shows that various Salmonella types such as

Enteritidis, Typhimurium, and Heidelberg, Tennessee, and Listeria
monocytogenes involving peanut butter products can cause
foodborne illness.
There are a variety of methods to reduce the bacteria presence in
butter such as thermal inactivation, nonlethal doses of e-beam
irradiation, high-pressure processing, some effective some
ineffective, and the usage of probiotics.

References
1. Dong HJ, Cho S, Boxrud D, et al. Single-nucleotide polymorphism
typing analysis for molecular subtyping of Salmonella Tennessee isolates
associated with the 2007 nationwide peanut butter outbreak in the United
States. Gut Pathog. 2017 May 1;9:25.
2. Wilson MR, Brown E, Keys C, et al. Whole genome DNA sequence
analysis of Salmonella subspecies Enterica serotype Tennessee obtained
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from related peanut butter foodborne outbreaks. PLoS One. 2016;

11(6):e0146929.
3. Shachar D, Yaron S. Heat tolerance of Salmonella enterica serovars
Agona, Enteritidis, and Typhimurium in peanut butter. J Food Prot.
2006;69(11):2687-91.
4. He Y, Guo D, Yang J, e al. Survival and heat resistance of Salmonella
enterica and Escherichia coli O157:H7 in peanut butter. Appl Environ
Microbiol. 2011;77(23):8434-8.
5. Lathrop AA, Taylor T, Schnepf J. Survival of Salmonella during
baking of peanut butter cookies. J Food Prot. 2014;77(4):635-9.
6. Burnett SL, Gehm ER, Weissinger WR, Beuchat LR. Survival of
Salmonella in peanut butter and peanut butter spread. J Appl Microbiol.
2000;89(3):472-7.
7. Grasso EM, Somerville JA, Balasubramaniam VM, Lee K. Minimal
effects of high-pressure treatment on Salmonella enterica serovar
Typhimurium inoculated into peanut butter and peanut products. J Food
Sci. 2010;75(8):E522-6.
8. Matak KE, Hvizdzak AL, Beamer S, Jaczynski J. Recovery of
Salmonella enterica serovars Typhimurium and Tennessee in peanut
butter after electron beam exposure. J Food Sci. 2010;75(7):M462-7.
9. Ma L, Zhang G, Gerner-Smidt P, et al. Thermal inactivation of
Salmonella in peanut butter. J Food Prot. 2009;72(8):1596-601.
10. Kottapalli B, Nguyen SPV, Perez T, Cunningham A. Thermal
Inactivation of Salmonella and Listeria monocytogenes in Peanut butter-
filled pretzels and whole wheat pita chips. J Food Prot. 2019;82(2):238-46.
11. Klu YA, Chen J. Influence of probiotics, included in peanut butter,
on the fate of selected Salmonella and Listeria strains under simulated
gastrointestinal conditions. J Appl Microbiol. 2016;120(4): 1052-60.

VARIOUS POLUTANTS
Weiss & al. (1) mentioned that the main source of human
exposure to Persistent organic pollutants (POPs) is, in general, food.
In this study, 64 butter samples from 37 countries were analyzed to
assess the global contamination of polychlorinated dibenzo-p-dioxins
(PCDDs), dibenzofurans (PCDFs), biphenyls (PCBs),
hexachlorobenzene (HCB), and 2,2-bis (4-chlorophenyl)-1,1,1-
trichloroethane (DDT) together with its major metabolites. The
objectives of the study were to assess the presence of major
organohalogen contaminants in butter, to trace geographical
differences, and to determine toxic equivalents (TEQs) of PCDDs/Fs
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and dioxin-like PCBs in butter. The highest PCDD/F concentrations

were found in butter from Korea with an average of 1.4 pg TEQ g(-1)
lipid weight (l.w.). from PCDD/F and an additional contribution from
the non- and mono-ortho-PCBs of 0.55 pg TEQ g(-1) l.w. Belgian
butter showed average levels of 0.53 and 1.2 pg TEQ g(-1) l.w. for
PCDDs/Fs and PCBs, respectively, but one sample of Belgium butter
had a total TEQ level as high as 4.0 pg TEQ g(-1) l.w. Three out of five
butter samples from Portugal showed similarly high PCDD/F TEQ
levels. The sigmaPCB levels in European butter appeared to be
somewhat higher than in the samples from the rest of the world. The
average contribution of CB-153 to the total PCB concentration was
22% (SD 6.4, coefficient of variation 29%). Generally, the PCBs
contributed around 60% of the total TEQ value, with CB-126
contributing approximately half of this value. This shows the
important TEQ contribution from dioxinlike PCBs to the total TEQs.
The highest HCB levels were found in butter samples from Russia,
Ukraine, Belgium, and Slovenia. Low levels of HCB in butter were
generally found in the Southern Hemisphere. Butter samples from
countries from Eastern Europe had elevated sigmaDDT
concentrations, with a particularly high concentration in Ukraine
butter, followed by some Russian samples, Brazil, and the U.S. (1).
According to CDC (2), On November 25, 2008, an epidemiologic
assessment began of a growing cluster of Salmonella serotype
Typhimurium isolates that shared the same pulsed-field gel
electrophoresis (PFGE) pattern in PulseNet. As of January 28, 2009,
529 persons from 43 states and one person from Canada had been
reported infected with the outbreak strain. This report is an interim
summary of results from ongoing epidemiologic studies and recall
and control activities by CDC, the Food and Drug Administration
(FDA), and state and local public health agencies. Confirmed,
reported onset of illness dates have ranged from September 1, 2008,
to January 16, 2009. A total of 116 patients were hospitalized, and
the infection might have contributed to eight deaths. Sequential
case-control studies have indicated significant associations between
illness and consumption of any peanut butter (matched odds ratio
[mOR] = 2.53), and specific brands of prepackaged peanut butter
crackers (mOR = 12.25), but no association with national brand jarred
peanut butter sold in grocery stores. Epidemiologic and laboratory
findings indicate that peanut butter and peanut paste produced at
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one plant are the source of the outbreak. These products also are
ingredients in many foods produced and distributed by other
companies. This outbreak highlights the complexities of "ingredient-
driven" outbreaks and the importance of rapid outbreak detection
and investigation. Consumers are advised to discard and not eat
products that have been recalled (2).
According to CDC (3), in 2012, CDC collaborated with state health
and agricultural agencies and the Food and Drug Administration
(FDA) to investigate an outbreak of Salmonella Bredeney infections
associated with exposure to peanut products manufactured by
Sunland, Inc. of Portales, New Mexico (3).
Santillo & al. (4) reported that the potential for use of butter as a
widely available, relatively uniform lipid-rich matrix for the
determination of spatial distributions of persistent organic pollutants
has already been demonstrated. The present study determines the
contributions to toxicity equivalence (TEQ) from polychlorinated
dioxins and furans (PCDD/Fs) and polychlorinated biphenyls (PCBs)
using butter samples from 24 countries worldwide. Concentrations
of PCDD/Fs and dioxin-like PCBs ranged from 0.07 to 5.69 pg
SigmaWHO-TEQ g(-1) lipid. For most samples, PCDD/F TEQ fell within
ranges reported for European dairy products over the last decade
(0.3-2 pg x g(-1) lipid I-TEQ), though a single sample from Spain was a
notable exception. Other than this sample, the highest values were
recorded for samples from the Netherlands and Italy, with those
from India, China and Tunisia also being relatively high. The
contribution from non-ortho-PCBs was particularly significant in
samples from Germany, Austria, Italy, the Czech Republic, Tunisia,
India and Argentina. Although overall TEQs were generally highest in
European and Mediterranean butters, elevated levels were also
apparent in industrializing regions of Asia (India, China) and Latin
America (Argentina). More detailed regional studies would be
necessary to identify likely dioxin and PCB sources in each case.
Nevertheless, this study supports the utility of butter as a monitoring
matrix that may be especially applicable in regions for which
monitoring programs are currently lacking (4).
Waliszewski & al. (5) analyzed organochlorine pesticide residues
in 345 samples of butter purchased from Mexican supermarkets in
1994. Three national brands and one foreign brand were analyzed.
Most samples contained residues of gama-HCH (91%), HCB (90%),
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and p,p'-DDE (88%). Residues of alpha-HCH (63%), p,p'-DDT (42%),

beta-HCH (38%), o,p'-DDT (17%), heptachlor epoxide (7%), and
endosulfane sulfate (3%) were also detected. Mean values of
pesticide residues determined were 0.093 mg/kg fat for total HCH
and 0.056 mg/kg for total DDT. Mean values of organochlorine
pesticide residues in Mexican butter were comparable with those in a
foreign bran (5).
Schecter & al. (6) reported the first known incidence of U.S.
butter contamination with extremely high levels of polybrominated
diphenyl ethers (PBDEs). Ten butter samples were individually
analyzed for PBDEs. One of the samples and its paper wrapper
contained very high levels of higher-brominated PBDEs. Dietary
estimates were calculated using the 2007 U.S. Department of
Agriculture Loss-Adjusted Food Availability data, excluding the
elevated sample. The highly contaminated butter sample had a total
upper bound PBDE level of 42,252 pg/g wet weight (ww). Levels of
brominated diphenyl ether (BDE)-206, -207, and -209 were 2,000,
2,290, and 37,600 pg/g ww, respectively. Its wrapping paper
contained a total upper-bound PBDE concentration of 804,751 pg/g
ww, with levels of BDE-206, -207, and -209 of 51,000, 11,700, and
614,000 pg/g, respectively. Total PBDE levels in the remaining nine
butter samples ranged from 180 to 1,212 pg/g, with geometric mean
of 483 and median of 284 pg/g. Excluding the outlier, total PBDE
daily intake from all food was 22,764 pg/day, lower than some
previous U.S. dietary intake estimates. The data show that Higher-
brominated PBDE congeners were likely transferred from
contaminated wrapping paper to butter. A larger representative
survey may help determine how frequently PBDE contamination
occurs. Sampling at various stages in food production may identify
contamination sources and reduce risk (6).
Roszko & al. (7) reported that various statistical methods have
been employed to analyze in details seasonal diversification of
polychlorinated biphenyl (PCB)/polybrominated diphenyl ether
(PBDE) congener profiles found in butter fat. The variability of the
PCB/PBDE congener profiles indicates the presence of various
sources of the milk fat contamination. The obtained results suggest
that the environmental chemical background has the highest share in
the contamination sources pattern. Ion trap mass spectrometry
coupled to high-resolution gas chromatography with semi-permeable
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membrane dialysis sample cleanup was used for determination of

PCBs and PBDEs in milk fat. Determined butter fat PCB profiles were
similar to the profiles characteristic for Aroclor 1254 technical
mixture. The data indicate that dietary intake of PCB/PBDE with milk
and milk products may be estimated to be about 717.5 pg kg b.w.(-1)
day(-1) for six-indicator PCBs, 0.329 (7).
Aksoy & al. (8) evaluated the levels of 9 organochlorine
compounds (aldrin, hexachlorobenzene, 2,4-DDE, 4,4-DDE, 2,4-DDT,
4,4-DDT, and α-, β-, and γ-HCH) in butter samples collected in the
Eastern, Middle and Western Black Sea Regions of Turkey between
October 2009 and June 2010. The liquid-liquid extraction method
was used to extract the organochlorine compounds from the samples
and the measurements were performed by using a gas
chromatograph-electron capture detector system. DDT metabolites,
aldrin, hexachlorobenzene (HCB), and α-, and γ-HCH were not
detected in the samples but β-HCH was detected in 3 of a total of 88
samples. In the first period, only one sample from the West Black
Sea Region was β-HCH positive (0.014 mg kg(-1)). The other β-HCH
positive samples collected in Middle and West Black Sea Regions in
the second period had a concentration of 0.066 and 0.019 mg kg(-1),
respectively. All concentrations of the detected compounds
exceeded the legal limits of 0.003 mg kg(-1) for β-HCH, as prescribed
by the Turkish Food Codex, and therefore pose a potential health risk
for consumers. The contamination detected is most likely due to the
past usage of β-HCH in agriculture and its long term persistence in
the environment. These results strongly suggest that further
research should be focused on the detection of pesticide residues in
agricultural areas across the nation (8).
Dervisoglu & al. (9) purchased 88 randomly selected samples of
butter produced in the Black Sea region of Turkey from different
retail markets during different periods and investigated for toxic and
essential elements content. Quantitative analyses of elements in the
samples were performed using an inductively coupled plasma-mass
spectroscopy (ICP-MS). Mean concentrations of As, Cr, Cu, Fe, Mn,
Ni, Pb, Se and Zn in the butter samples were 18.93, 100.32, 384.66,
4199.1, 887.47, 168.64, 56.13, 16.34 and 384.66 µg kg(-1),
respectively. Cd and Co were detected in 19 (mean content 0.29 µg
kg(-1)) and 81 (mean content 3.81 µg kg(-1)) samples of 88 butter
samples, respectively. However, the dietary intake of these elements
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by the population of the Black Sea region is currently well below the
dietary reference intake (DRI) and provisional tolerable weekly intake
(PTWI) levels of essential and toxic elements (9).
Lenardón & al. (10) analyzed organochlorine pesticide residues in
150 samples of butter collected from several places in the cities of
Santa Fe and Rosario (Argentine Republic) over a period of 18
months. Pesticides analyzed in butter were: HCH (alpha and gamma
isomers), heptachlor and its epoxide, aldrin, dieldrin, and DDT (p,p'
DDE, o,p' DDT and p,p' DDT). Most samples contained residues of
gamma-HCH (Lindane) and heptachlor (92% and 78%, respectively);
alpha-HCH and aldrin were detected in 58% and 55% of samples;
dieldrin, heptachlor epoxide, DDT isomers were found in few cases
(30%). Mean values of pesticides residues in butter fat were: sigma
HCH 0.029 ppm; sigma heptachlor 0.064 ppm; sigma aldrin 0.11 ppm
and sigma DDT 0.024 ppm. Residue levels exceeded the limits
prescribed by the Food and Agricultural Organization/World Health
Organization in only very few cases (10).
Bedi & al. (11) monitored organochlorine, organophosphate, and
synthetic pyrethroid pesticide residues in butter (n=55) and ghee
(n=56) samples collected from three different regions of Punjab. The
estimation of pesticide residues was done by multiple residue
analytical technique using gas chromatography equipped with GC-
ECD and GC-FTD. The confirmation of residues was done on gas
chromatography mass spectrometry in both selective ion monitoring
(SIM) and scan mode. Results indicated the presence of
hexacholorocyclohexane (HCH) and p,p' DDE as predominant
contaminant in both butter and ghee. Residues of HCH were
detected in 25 and 23% samples of butter and ghee, respectively,
while residues of p,p' DDE were recorded in 29 and 25% of butter and
ghee samples, respectively. None of the butter and ghee sample
violated the MRL values of 200 ng g(-1) for HCH and 1250 ng g(-1) for
dichorodiphenyl tricholorethane (DDT). The presence of endosulfan,
cypermethrin, fenvalerate, deltamethrin, and chlorpyrifos were
observed in a few butterand ghee samples at traces. The spatial
variation for comparative occurrence of pesticide residues indicated
higher levels in the south-western region of Punjab. Additionally, the
temporal variation indicated the significant reduction of HCH and
DDT levels in butter and ghee in Punjab (11).
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Kumari & al. (12) collected butter (45) and ghee (55) samples
from rural and urban areas of cotton growing belt of Haryana and
analyzed for detecting the residues of organochlorine, synthetic
pyrethroid and organophosphate insecticides. The estimation was
carried out by using multi residue analytical technique employing GC-
ECD and GC-NPD systems equipped with capillary columns. Butter
samples were comparatively more contaminated (97%) than ghee
(94%), showing more contamination with organochlorine insecticides
from urban samples. About 11% samples of butter showed
endosulfan residues above MRL value and 2% samples had residues
of synthetic pyrethroids and organophosphates each above their
respective MRL values. In ghee, residues of HCH & DDT both and of
endosulfan exceeded the MRL values in 5 and 20% samples,
respectively. Among organophosphates, only chlorpyriphos was
detected with 9% samples showing its residue above MRL value.
Irrespective of contamination levels, residues above the MRL values
were more in ghee. More extensive study covering other agricultural
regions/zones of Haryana has been suggested to know the overall
scenario of contamination of milk products (12).
Njoroge & al. (13) conducted a 3-year comprehensive analysis of
aflatoxin contamination in peanut butter in Zambia, sub-Saharan
Africa. The study analyzed 954 containers of 24 local and imported
peanut butter brands collected from shops in Chipata, Mambwe,
Petauke, Katete, and Nyimba districts and also in Lusaka from 2012
to 2014. For analysis, a sample included six containers of a single
brand, from the same processing batch number and the same shop.
Each container was quantitatively analyzed for aflatoxin B1 (AFB1) in
six replicates by using competitive enzyme-linked immunosorbent
assay; thus, aflatoxin contamination level of a given sample was
derived from an average of 36 test values. Results showed that 73%
of the brands tested in 2012 were contaminated with AFB1 levels >20
μg/kg and ranged up to 130 μg/kg. In 2013, 80% of the brands were
contaminated with AFB1 levels >20 μg/kg and ranged up to 10,740
μg/kg. Compared with brand data from 2012 and 2013, fewer brands
in 2014, i.e., 53%, had aflatoxin B1 levels >20 μg/kg and ranged up to
1,000 μg/kg. Of the eight brands tested repeatedly across the 3-year
period, none consistently averaged ≤20 μg/kg. The survey clearly
demonstrates the regular occurrence of high levels of AF B1 in
peanut butter in Zambia. Considering that some of the brands tested
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originated from neighboring countries such as Malawi, Zimbabwe,

and South Africa, the current findings provide a sub-Saharan regional
perspective regarding the safety of peanut butter (13).
Elzaput & al. (14) analyzed 43 peanut butter samples from
Khartoum State, Sudan, for aflatoxins (AFs, AFB1 + AFB2 + AFG1 +
AFG2) using high performance liquid chromatography (HPLC) with
fluorescence detection after extraction with methanol: water (8:1,
v/v) and clean-up using chloroform. All samples were contaminated
with AFs, with total AF levels ranging between 26.7 and 853 μg/kg,
and a mean total AF level of 287 ± 200.5 μg/kg. The highest
concentrations were found for AFB1, (28 positive samples, maximum
534 μg/kg), while AFG1 was most frequently detected (43 positive
samples, maximum 401 μg/kg). AFB2 (42 positive samples, maximum
3.2 μg/kg) and AFG2 (4 positive samples, maximum 30 μg/kg) were
also present in these samples. The mean AF contamination levels
found in this study exceeded by far all international regulations
concerning maximum levels for this group of toxins. The data show
that the levels of AF contamination in peanut butter from the
Kartoum area are quite alarming, and may pose serious health
hazards to consumers. Therefore, an intervention strategy to
manage AF in peanut butter is urgently needed (14).
Elshafie & al. (15) carried out a survey to detect the presence of
aflatoxin B(1) in 60 duplicated samples (120 samples) of peanuts
butter purchased from the local markets and other traditionally
prepared and distributed by the street sellers in Khartoum state,
Sudan. AflaTest-P affinity column was used to extract the toxin from
the samples, and the concentration was measured by calibrated
Vicam fluorometer. Aflatoxin B(1) was detected at variable levels in
100% of the screened samples. Traditionally prepared samples
showed the highest incidence of aflatoxin B(1) which is above the
internationally regulated tolerance levels (5-20 ppb). The means and
the ranges of the aflatoxin B(1) recovered were as follows: 63.9 ppb
(29-128 ppb), 54.5 ppb (21-131 ppb) and 101 ppb (17-170 ppb) for
samples collected from Khartoum, Khartoum North and Omdurman
areas, respectively. Samples from retail stores presented relatively
low aflatoxin B(1) incidences 14.5 ppb (1-57 ppb), but only 30% of the
samples revealed aflatoxin level below 10 ppb. Laboratory
segregated and carefully prepared butter from good grade nuts
showed the lowest levels of this toxin (3.3 ppb; 2-6 ppb). The results
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showed that peanuts butter prepared by the street sellers and

distributed by the retail stores are evidently hazardous to human
health. There is therefore urgent need for strong form of quality
control measures and public awareness. The use of excellent grade
peanuts and care during processing and storage are priority (15).
Vega (16) developed a rapid extraction technique was developed
for the isolation and subsequent liquid chromatographic
determination of aflatoxins B1, B2, G1, and G2 in creamy and crunchy
peanut butter. Peanut butter samples were extracted with a
methanol 15% sodium chloride (7 + 3) solution followed by a second
extraction with methanol. The extract was subjected to a cleanup
using a Vicam Aflatest immunoaffinity column. Control samples for
both smooth and crunchy peanut butter were fortified at 4 different
levels for aflatoxin B1, B2, G1, and G2. The average aflatoxin B1, B2,
G1, and G2 recoveries from smooth peanut buffer were 95.2, 89.9,
94.1, and 62.4%, respectively, and 92.4, 84.3, 85.5, and 53.7%,
respectively, from crunchy peanut butter. This extraction method
and the official AOAC Method 991.31 produced comparable results
for peanut butter samples. This method provides a rapid, specific,
and easily controlled assay for the analysis of aflatoxins in peanut
butter with minimal solvent usage. Organic solvent consumption was
decreased by 85% and hazardous waste production was decreased by
80% in comparison with the AOAC method. Along with the
decreased solvent consumption, significant savings in time were
observed (16).

This chapter (1-16) shows that butter can be contaminated with a

variety of pollutants and carcinogenic aflatoxin that are dangerous to
human health.

References
1. Weiss J, Päpke O, Bergman A. A worldwide survey of
polychlorinated dibenzo-p-dioxins, dibenzofurans, and related
contaminants in butter. Ambio. 2005;34(8):589-97.
2. Centers for Disease Control and Prevention (CDC). Multistate
outbreak of Salmonella infections associated with peanut butter and
peanut butter-containing products - United States, 2008-2009. MMWR
Morb Mortal Wkly Rep. 2009;58(4):85-90.
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3. Centers for Disease Control and Prevention (CDC). Notes from the
field: Salmonella Bredeney infections linked to a brand of peanut butter -
United States, 2012. MMWR Morb Mortal Wkly Rep. 2013;62(6):107.
4. Santillo D, Fernandes A, Stringer R, et al. Butter as an indicator of
regional persistent organic pollutant contamination: further development
of the approach using polychlorinated dioxins and furans (PCDD/Fs), and
dioxin-like polychlorinated biphenyls (PCBs). Food Addit Contam. 2003;
20(3):281-90.
5. Waliszewski SM, Pardío VT, Waliszewski KN, et al. Levels of
organochlorine pesticides in Mexican butter. J AOAC Int. 1996;79(3):784-
6.
6. Schecter A, Smith S, Colacino J, et al. Contamination of U.S. butter
with polybrominated diphenyl ethers from wrapping paper. Environ
Health Perspect. 2011;119(2):151-4.
7. Roszko M, Szymczyk K, Rzepkowska M, Jędrzejczak R. Seasonal
variability of polychlorinated biphenyls (PCBs) and polychlorinated
diphenyl ethers (PBDEs) congener profiles in butter in Poland: dietary risk
evaluation. J Environ Sci Health B. 2014;49(3):182-99.
8. Aksoy A, Dervisoglu M, Guvenc D, et al. Levels of organochlorine
pesticide residues in butter samples collected from the Black Sea Region of
Turkey. Bull Environ Contam Toxicol. 2013;90(1):110-5.
9. Dervisoglu M, Gul O, Yazici F, et al. Toxic and essential elements in
butter from the Black Sea region, Turkey. Food Addit Contam Part B
Surveill. 2014;7(1):49-53.
10. Lenardón A, Maitre de Hevia MI, Enrique de Carbone S.
Organochlorine pesticides in Argentinian butter. Sci Total Environ. 1994;
144(1-3):273-7.
11. Bedi JS, Gill JP, Aulakh RS, Kaur P. Occurrence and spatial
distribution of pesticide residues in butter and ghee (clarified butter fat) in
Punjab (India). Environ Monit Assess. 2016;188(2):100.
12. Kumari B, Singh J, Singh S, Kathpal TS. Monitoring of butter and
ghee (clarified butter fat) for pesticidal contamination from cotton belt of
Haryana, India. Environ Monit Assess. 2005;105(1-3):111-20.
13. Njoroge SM, Matumba L, Kanenga K, et al. A case for regular
aflatoxin monitoring in peanut butter in Sub-Saharan Africa: lessons from
a 3-Year Survey in Zambia. J Food Prot. 2016;79(5):795-800.
14. Elzupir AO, Salih AO, Suliman SA, et al. Aflatoxins in peanut butter
in Khartoum State, Sudan. Mycotoxin Res. 2011;27(3):183-6.
15. Elshafie SZ, ElMubarak A, El-Nagerabi SA, Elshafie AE. Aflatoxin B1
contamination of traditionally processed peanuts butter for human
consumption in Sudan. Mycopathologia. 2011;171(6):435-9.
16. Vega VA. Rapid extraction of aflatoxin from creamy and crunchy
peanut butter. J AOAC Int. 2005;88(5):1383-6.
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SUMMARY
This study shows that butter, a delicious nutritional item, was
widely used on a variety of occasions in Biblical times.
Over the years, scientific study has revealed a variety of positive
and negative effects of butter. While there are numerous positive
effects, there are relatively few negative or neutral overall
associations of butter with conditions such as cardiovascular disease.
Moreover, higher peanut butter consumption has the potential
benefit of lowering the risk of type 2 diabetes in women.
Substituting butter and solid margarine with soft margarine for use as
spreadable fats may be associated with reduced risk of myocardial
infarction.
It is recommended that hypercholesterolemic individuals should
keep their consumption of butter to a minimum, whereas moderate
butter intake may be considered part of the diet in the
normocholesterolemic population.
Some negative effects include contamination with bacteria such
as types of Salmonella like Enteritidis, Typhimurium, and Heidelberg,
Tennessee, and Listeria monocytogenes, and contamination with
various toxic pollutants.
In spite of some negative effects, the ancient food of butter can
be considered a useful element of human nutrition.

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