Sagay National High School

Science Technology & Engineering

Lactobacillus from Milk, Rice and Sugar as Probiotic for the Growth Performance of Broiler Chicken

MA. VICTORIA A. JIMENEA

BLESSY GRACE G. OLASIMAN

PATRICK ANGELOU F. PEREZ

Researchers

Reynaldo L. Huervana Jr.

Research Adviser

Sagay National High School

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 TABLE OF CONTENTS

INTRODUCTION 1

 Background of the Study 2

 Statement of Objective 2

 Hypothesis 2

 Significance of the Study 3

 Scope and Limitation 3

 Definition of Terms 4

REVIEW OF RELATED LITERATURE 5-8

METHODOLOGY

 Equipment/materials 9

 Fermentation of lactobacillus 10

 Brooding 11

 Feeding and watering 12

 Sanitation 13

 Clearing-disposal of waste 13

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Chapter I

Introduction

Lactobacillus is a rod-shaped bacterium that produces lactic acid from the fermentation of carbohydrates. It

is a microorganism that can be used around the house, for animals and even for gardens. It is a genus of beneficial

bacteria, they are found everywhere and there are even some inside your digestive system. Lactobacillus bacteria

are in many probiotic supplements and lactobacillus species plays an important role in the production of many

common foods. In the garden, lactobacillus serum can be used as a digester, helping break down organic matter

and turn it into a form that is available to your plants. Lactobacilli were the first genus of bacteria proved to have

beneficial health effects. They have been shown to be present in the gastrointestinal tract of most animals and

birds. It is one of the many friendly species of intestinal microflora considered as beneficial bacteria in its ability

to aid in breakdown of proteins, carbohydrates and fats in food and help absorption of necessary elements and

nutrients. They were also referred to as “live enzyme factory” as they produce wide range of enzymes, which can

breakdown even complex carbohydrates, hence beneficial to the host (Anonymous 2002).

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 1.1 ‘Background of the Study

Probiotics are beneficial organisms like bacteria and yeast that are believed to improved health when

consumed. Both plants and animals need these beneficial microbes to stay healthy. Without the teeming good

bacteria in the intestine of man and animals, digestion will not occur. Probiotics are to reduce foul odor in

animal houses, homes, etc. It can also help to free clogged drains. The good bacteria in probiotics eat up the

bad bacteria that cause smell and diseases. The researchers conduct this study to find out if lactobacillus has an

effect to the growth of broiler chickens.

1.2 Statement of the Objective

 To determine the effect of lactobacillus as probiotic on broiler chicken

 To test how effective is the lactobacillus on broiler chicken

 To measure the weight of broiler chickens after feeding with lactobacillus

Hypothesis

There is no significant effect of Lactobacillus out of rice, milk and sugar as probiotic to the growth

performance of broiler chickens.

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1.3 Significance of the Study

The Philippine broiler industry is now faced with the challenge of beefing up production through importation

of chicken meat and chicken meat cuts. The apparent reduction of chicken meat consumption is brought by the

major outbreak of Highly Pathogenic Avian Influenza (HPAI) and Ebola Virus. These events caused the

exceedingly high prizes of broiler meat and a built up in inventory. This study may lead up to a better meat quality

in the country. This study can help chicken breeders to breed broilers healthier.

1.4 Scope and Limitation

This study will be conducted in Sagay National High School mini laboratory. The

lactobacillus culture will be made out of rice, milk and sugar for one week. The lactobacillus culture will be put

in a jar and will be use as probiotic to the growth if broiler chickens. The lactobacillus will be used as a probiotic

to enhance the growth of broiler chickens. The broiler chickens will stay in a chicken cage.
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Definition of Terms

Term Conceptual Definition Operational Definition

Lactobacillus The one that is being fed to A rod-shaped bacterium that

the broiler chickens produces a lactic acid from

the fermentation of

carbohydrates

Probiotic The use of lactobacillus Denoting a substance that

stimulates the growth of

microorganisms, especially

those with beneficial

properties

Broiler A young chicken suitable for A young chicken suitable for

roasting, grilling, barbecuing. roasting, grilling, barbecuing.

The one that is being fed by

lactobacillus

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Chapter II

Review of Related Literature

Metchinkoff (1907) suggested that the health status and longevity could be improved by consumption of

milk fermented by lactobacilli, as he observed it in some of the rural inhabitants of the Caucasus who consumed

fermented milk throughout their life (Mackenzie 1925).

Lilley and Stillwell (1965) first introduced the term “Probiotic” to described, “growth promoting factors”

produced by microorganisms. Parker (1974) used the term probiotics for microorganisms or substances that

contribute to intestinal microbial balance. Fuller (1989) redefined the probiotic as “a live microbial feed

supplement, which beneficially affects the host animal by improving the intestinal microbial balance”.

Havenaar et al. (1992) pointed out that the definition given by Fuller (1989) was restricted to feed

supplements, animals and the intestinal tract. Hence, they redefined the probiotic considering both man and

animals as a “mono or mixed culture of living microorganisms which beneficially affect the host by improving

the properties of indigenous microflora”.

Tortuero (1973) observed increased weight gain, better feed conversion and although not statistically

significant difference in fat digestibility and nitrogen retention in chicks fed with the probiotics.

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Lactic acid bacteria

Lactic acid bacteria (LAB) are group of Gram-positive bacteria that are devoid of cytochromes and preferring

anaerobic conditions, fastidious, acid-tolerant and strictly fermentative. They are usually non-motile and non-

sporulating bacteria that produce lactic acid. This bacterial group contains both rods (Lactobacilli and

Carnobacteria) and cocci (Streptococci). Different species of lactic acid bacteria (such as Streptococcus,

Leuconostoc, Pediococcus, Aerococcus, Enterococcus, Vagococcus, Lactobacillus, Carnobacterium) have

adapted to grow under widely different environmental conditions. They are found in the gastrointestinal tract of

various animals, dairy products, seafood products, soil and on some plant surfaces (Ring & Gatesoupe, 1998).

Although lactic acid bacteria are not dominant in the normal intestinal microbiota, several trials have been

undertaken to induce an artificial dominance of lactic acid bacteria (Verschuere er al., 2000). Based on theie

carbohydrate metabolism LAB are divided into two distinct groups. The homo-fermentative group utilizes he

Embden-Meyerhof-Parnas (glycolyctic) pathway to transform a carbon source chiefly into lactic acid. Hetero-

fermentative bacteria produce equimolar amounts of lactate, carbon dioxide, ethanol or acetate from glucose

exploiting phosphoketolase pathway. Homo-fermentative group consist of Lactococcus, Pediococcus,

Enterococcus, Streptococcus. Hetero-fermentative group include Leuconostoc, Weisella (Vasiljevik &Shah,

2008)

In commercial broiler production, Couch (1978) recorded an increase in average body weight of 47 grams

by using lactobacillus supplements at a rate of 454 g. per ton of feed.

Adler and DaMassa (1980) reported that feeding a lactobacillus culture to chicks resulted in an

improvement in body weights and reduced the occurrence of pasted vent.

Contrary to the reports of various beneficial responses of probiotic supplementation in broiler diet,

Bruenrostra and Kratzer (1983) found that the lactobacillus supplement group of broiler chicken did not show as

much as good result compared to antibiotic supplemented and control groups.

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 Watkins and Kratzer (1983) recommended that there was possibly an optimum level of lactobacilli

required by the chicken that provided most of the benefits to the host. Inclusion of the lactobacilli above the

recommended levels resulted in undesired effect such as bacterial competition for biotin and low levels were not

enough to show the desired benefits.

As reported by Jernigan et al. (1985) the research related to the use of lactic acid producing bacteria as

growth stimulants in broilers was not sufficient. According to them, lactobacilli had an advantage that the

Lactobacillus species were capable of producing large amount of lactate from simple carbohydrates, can withstand

a high degree of acidity which was usually fatal to other bacteria and have long been considered as desirable

inhabitants of digestive tract.

Franceschi and Stocker (19910 reviewed the mechanism of action with respect to the beneficial effects of

lactic acid producing bacteria on digestive system in broilers and layers.

Supplementation of lactobacillus cultures at the level of 0.5 gram per kg. of broiler starter increased daily

weight gain and feed conversion efficiency and decreased the incidence of diarrhea and mortality in broiler

chicken (Hussein and Ashrey, 1991).

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 Review of Related Studies

In 2016, Professor Dr. S. M. Lutfur Kabir in his study on isolation, identification, molecular

characterization of screening of probiotic activities of Lactobacillus species from poultry sources at live bird

markets in Mym ensingh, Bangladesh, found out that the results of antibiotic sensitivity showed that all of the

isolates tested were sensitive to tetracycline and gentamlycin. However, most of the isolates tested were resistant

to ampicillin and cefradine. Potential probiotic bacterial stains from poultry sources were identified as well as the

characterized in the study.

Kabir conducted a study to isolate and characterized Lactobacilli from the gastrointestinal (G1) tract of

the broiler chickens in different markets along with evaluation of probiotic ability and antibiotic sensitivity of

Lactobacillus isolates during the period of July, 2014 to December 2014. The caecum and cloacal contents of 100

broiler chickens from different markets were collected and cultured on Man Rogosa Sharp (MRS) both and agar.

Lactobacilli were isolated and characterized by using phenotypic (cell morphology, Gram’s staining.

Physiological and biochemical tests which are specific for Lactobacillus genus) and genotypic methods (PCR and

RAPD). The identified Lactobacilli species were screened for probiotic properties by in vitro tests like acid

tolerance and bile tolerance. Total eighty-two isolates were identified as Lactobacillus based on morphological,

physiological and biochemical tests which are specific for Lactobacillus genus. All of the Lactobacillus isolates

were amplified by using 16S RNA gene-based universal primers. Furthermore, out of 10 Lactobacillus isolates 4

molecular patterns method in this study. Out of 15 Lactobacillus isolates, 10 isolates demonstrated probiotic

ability as determined by bile tolerance test and acid tolerance test.

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Chapter III

Methodology

A. Equipment/materials

The following materials were used in the study. 20 pcs. chicks, water, rice, sugar, milk, glass jars,

strainer, airlock or 1-way venting lid for your container, 3 incandescent bulbs (100 watts each), newspapers.

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 B. Fermentation of lactobacillus

Gather the wild bacteria. Rinse a cup or sow of rice in a few cups of water. The water

will get cloudy as it picks up starches from the rice. Take the dish of starchy water. Leave it for 3-7 days.

Bacteria from the environment will be attracted to the starchy water and will colonize the liquid in the dish.

After a few days, the liquid will start to separate into 3 distinct layers. The middle layer is what the researchers

want. Separate it from the rest and move it to a larger jar or container. Add milk so the lactobacillus takes over

the culture. In a large container, combine about 10 parts of milk to 1 part of the native bacteria culture. Leave it

undisturbed for 5-10 days at room temperature. After few days, the bacteria and milk mixture will start to

separate into layers. The researchers are after the liquid layer. Add sugar to the lactobacillus culture to preserve

and stabilize it. Mix the lacto culture with a roughly equal amount of sugar and leave it for 2-3 weeks.

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 C. Brooding

All of the chicks were placed inside a “brooder”, covered with cardboard and floored

with 15 layers of used newspaper that serves as feeding area fro few days. One incandescent bulb (100 watts)

was placed inside the “brooder” which will stimulate heat during brooding to meet the needed heat for the

chicks to survive. The brooding stage lasted for 10 days. The “brooder’s” temperature was regulated

beforehand.

Feed the chicks with water mixed with sugar. Two linear feeders with chick booster mash were provided

to the chicks. At the start a few of feeds were scattered in the newspaper flooring for the chicks to familiarize

the feeding area.

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 D. Feeding and Watering

The chicks were provided with free access to food and water. The chicks were fed with

commercially available feed (B-meg: Chick Booster Mash). Water that was provided to each cages varied

differently, the experimental group were watered with a mix of distilled water and fermented lactobacillus using

the ratio of [1:15]. The control group was provided with ordinary tap water. Water was changed every morning.

Amount of water consumed by each group was measured daily.

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 E. Sanitation

Clearing and cleaning of dirt and droppings was done every 3 days. Equipment was cleaned

after every use. Sanitation include: cleaning of cages, groundwork, surroundings, cleaning and clearing

hindrances and problems that may threat the growth of chicks. The surrounding was regularly checked and

cleaned.

F. Clearing/Disposal of Waste

The droppings and dirt were used as an organic fertilizer to the existing plants.

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References

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ltemid=110

http://shodhganga.inflibnet.ac.in/bitstream/10603/5613/9/09_chapter%202.pdf

http://shodhganga.inflibnet.ac.in/bitsream/1063/6920/6/06_chapter%202.pdf

FarmersJoint.com

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www.pchrd.dost.gov.ph

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