Digestion and absorption of nutrients

Dr. Shaharior Hashem

Associate Professor
Department of Aquaculture
Bangladesh Agricultural University
 ❑Outline of Presentation

• Introduction

• Digestive fluids and enzymes

• Absorption and assimilation of protein, fat and carbohydrate

• Rate of digestion, apparent and true digestibility

• Direct and indirect methods of digestibility determination

• Factors affecting digestibility

Introduction
 Digestion
• Process where ingested food nutrients (proteins, lipids and complex
carbohydrates) are broken down into units that are small enough to be
absorbed across the gut wall.

• Accomplished through to the action of digestive enzymes.

• Fish food digest ability depends on the secretion of quantities of

enzymes.

• Enzymes are stored in an inactive or proenzyme form.

• Inactive enzymes are converted to the active form which is influenced

by pH.
 Absorption
• Several mechanisms- Simple diffusion, active transport, pinocytosis.

• In Simple diffusion, solutes pass through the membrane from an

environment of high to low solute concentration without using energy.

• Active transport requires a continuous supply of energy and transport

solutes only in one direction from low to high solute concentration.

• Carrier system that utilizes Na+ and ATPase activity is needed for the
active transport of glucose and some amino acids.

• Pinocytosis also known as fluid endocytosis and bulk-phase pinocytosis.

Different absorption
 Pinocytosis
• Materials are taken into the cell through an invagination and
subsequent dissolution of a part of the cell membrane.

• Enables the cell to absorb some proteins and lipid in intact form.
Digestive fluids and enzymes
 Digestive fluids and enzymes
• Part of gastric secretions, pancreatic secretions, bile and intestinal
secretions.

• Acid gastric fluid production probably occurs in most fish, except in

those with out a stomach when neither HCl nor pepsin is formed in the
gut.

1. Stomach (gastric secretions):

Enzyme: HCl

Functions: reduce gut pH and allows pepsinogens to act.

 Digestive fluids and enzymes
2. Gastric glands:

Enzyme:
• Zymozen

Functions: Proteolytic enzymes (break down protein)

• Pepsinogen, HCl, pepsin

Functions: Cleave peptide linkages at the NH2 groups of aromatic and
acidic amino acids. Attack most proteins.
• Amylase, Lipase, Esterases, Chitinase
Functions: Acts on carbohydrates, lipids, esters, chitin
 Digestive fluids and enzymes
3. Pancreas:

• Enzymes
Functions: stores as zymogens. Proteases produced by the intestine
convert trypsinogen into trypsin.
• Bicarbonate
Functions: Neutralizes HCl entering intestine and prepare intestine for
alkali digestion.
• Proteases (trypsin, chymotripsin, carboxypeptidases and elastase)
Functions: Optimal action of enzymes at pH 7.0

• Chymotrypsin
Functions: Attacks peptide with carboxyls from aromatic side chains.
 Digestive fluids and enzymes
• Trypsin
Functions: Peptide linkages at carboxyl groups of lysine or arginine.
• Elastase
Functions: Attacks peptide bonds on elastin.
• Carboxy peptidases
Functions: Hydrolyses the terminal peptide bond of their substrates.

• Amylase
Functions: Carbohydrate digestion at nonacidic pH.

• Lipases
Functions: Hydrolyse triglycerides, fats, phospholipids.
 Digestive fluids and enzymes
4. Bile (secreted by the liver):

• Bile salts, cholesterol, phospholipids, inorganic ions

Functions: Make the intestinal medium alkaline, emulsify lipids.
5. Intestinal enzymes:

• Aminopeptidases, Polynucleotidase, Lecithinase

Functions: Split nucleosides, split nucleic acids, splits phospholipids into
glycerol, fatty acids.

• Various carbohydrate digesting enzymes

Functions: split carbohydrate.
Absorption and assimilation of protein, fat and
carbohydrate
 Digestion and absorption of Proteins
1. Pepsin:

• Play important roles in protein digestion in fish stomachs.

• pH optimum for pepsin is about pH 2.0

• Synthesized in the gastric gland in the inactive form called pepsinogen.

• HCl converts the inactive pepsinogen to the active pepsin.

• Pepsin is an endopeptidase and cleaves or cuts most peptide bonds in

amino acids and acidic amino acids.

 Digestion and absorption of Proteins
2. Trypsin and Chymotrypsin:

• Involved in the alkaline digestion of proteins.

• These enzymes are synthesized, stored and secreted in an inactive form

by the pancreas and transported to the midgut.

• Trypsinogen is activated in the intestine by enterokinase, an enzyme

selected from the intestinal mucosa which activated trypsinogen

become trypsin.
 Digestion and absorption of Proteins

• Both trypsin and chymotrypsin are endopeptidases but cleave different

linkages in a protein.

• Trypsin cleaves peptide linkages which are formed by basic amino

acids, arginine, lysine, and histidine.

• PEPSINOGEN TRYPSINOGEN CHYMOTRIPSINOGEN

HCl enterokinase trypsin

PEPSIN TRYPSIN CHYMOTRIPSIN

 Digestion and absorption of Proteins
3. Carboxypeptidases:

• Also secreted from the pancreas in the inactive form.

• These are exopeptidases (Carboxypeptidases A and Carboxypeptidases B) which

cleave the C-terminal amino acid of peptides or proteins.

• Carboxypeptidases A is active towards proteins with aromatic C-terminal amino

acids (phenylamine, tyrosine, and tryptophan).

• Carboxypeptidases B acts preferentially on those peptides with basic amino acids

(lysine and arginine).

 Digestion and absorption of Proteins
4. Aminopeptidases:

• Exopeptidases that act on N-terminal peptide of proteins.

• Protein molecule is first hydrolysed into the relatively large polypeptide

fragments by endopeptidases.

• Fragments are then hydrolyzed by enzymes acting on the amine and

carboxyl bonds and finally dipeptidases reduce the protein to its amino

acids.

• Proteins can be absorbed as whole proteins, peptides or free amino acids.

 Digestion and absorption of Carbohydrates

• It digest complex carbohydrates and are found in fish intestine.

• The enzyme amylase catalyses the digestion of starch.

• Two forms: α- amylase and β-amylase.

• α- amylase which acts randomly cleaving the chain form.

• β-amylase which cuts the chain at every two glucose unit.

• At the branched point, another enzyme, dextrinase does the work.

• The amylase and dextrinase produce maltose.

 Digestion and absorption of Carbohydrates

• Maltase hydrolyses maltose to give glucose, the final product of starch

digestion.

• On milkfish (an omnivore), extracts from the intestines, pancreas and liver

showed high levels of amylase activity.

• Maltose, trehalose, dextrin, starch and glycogen are rapidly hydrolysed in

the presence of crude extracts from the intestines and pyloric caeca of

milkfish.
 Digestion and absorption of Carbohydrates
• Other complex carbohydrates that are potential sources of energy but are not
readily digested are cellulose.

• Cellulose is mediated by two enzymes: cellulase and cellobiase.

• Cellulase hydrolyses cellulose to disaccharide cellobiase, which is then acted
upon by cellobiase producing the final breakdown product, glucose.

• Cellulytic bacteria are widely distributed in nature.

• Carbohydrate absorption has been tested by measuring the uptake of glucose,
the usual final end product of carbohydrate digestion.
• Glucose transport appears to be lowest in carnivores and highest in herbivores.
 Digestion and absorption of Lipids
• The liver plays a very important role in fat digestion.
• The bile that is produced in the liver and stored in the gall bladder is released
when food arrives in the intestine.
• All fat digesting enzymes are classed as lipolytic enzymes or lipases.
• Lipases hydrolyze ester linkages in triglycerides.

• The end products of lipase activity are glycerol and fatty acids.

• Lipases are detected in the pancreases, pyloric caeca, intestine, and liver.

• Phospholipases are enzymes that hydrolyze phospholipids.

 Digestion and absorption of Lipids
• Major difference between fat hydrolysis and that of protein or carbohydrates
is that lipases show relatively little substrate specificity and many will
catalyse hydrolysis of almost any organic ester.
• The other differences between lipases and proteases and carbohydrates is that
lipases fairly readily catalyse the synthesized fat and other esters.
• Lipids are taken up by the intestinal, partly as fatty acids, mostly as mono-
glycerides and partly as droplets, and transferred into blood and lymph
vessels.
• The lipid absorption process in fish, although much slower than in mammals,
does not differ fundamentally.
Rate of digestion, apparent and true digestibility
 Rate of Digestion
• Digestion rate is the time it takes to digest the food.
• Gut transit time- Time interval between ingestion and first appearance of
faces.
• Gut evacuation time- Time taken for the entire quantity of food ingested to be
voided.
• Gastric evacuation time- Time taken between ingestion and emptying of the
stomach.
• Retention time- The difference between evacuation time and transit time is
referred to as the retention time.
 Methods to measure Rate of Digestion

1. Sacrifice Method
• Fish are fed a predetermined quantity of food, sacrificed at predetermined
interval and the amount of food remaining in the stomach are estimated.
• Samples are obtained periodically by serial slaughter to determine gastric
emptying rate.
• The amount remaining in the stomach can be estimated as a percentage of the
volume, weight or in calorific value of the amount ingested.
• One limitation of this method is that it assumes that all the fish ate exactly the
same amount.
 Methods to measure Rate of Digestion
2. X- radiography method
• Most commonly used method is to incorporate an isotope into the feed.
• This method generally involves considerable stressing of the fish, including
constant handling and force feeding.
3. Use of dyes
• Incorporated into diets and the time at which the dye appears first in the faeces
is determined.
4. Direct observation
• Applicable in larval stage when the gut are visible in transparent larvae.
• Fish tend to feed continuously, it is helpful to label a quantum of the diet in such
a way that this portion of the meal is detectable in faces.
Direct and indirect methods of digestibility
determination
 Determination of Digestibility
Direct determination
• The quantity ingested and faecal matter voided is determined.
• It is difficult to determine the quantity ingested and the total amount of faeces
voided accurately as such determination are subjects to many errors.
• Indirect determination
• First used by Swedish scientist Edin in 1918.
• A marker is a material, though to be ingestible, which is introduced in small
quantities and distributed evenly in the test diet.
• A marker should not influence the physiology of digestion of the experimental
animal, should move along the gut at the same rate as the rest of the food material
and should not be toxic.
 Determination of Digestibility

• Commonly used external markers, those introduces into the diet are Cr2O3
(Chromic Oxide) and polypropylene.

• By far the most commonly used external marker is Cr2O3 in fish as well as in
terrestrial animal.

• Endogenous or internal markers commonly utilized for digestibility estimations

are crude fiber, hydrolysis resistant organic matter (HROM) and hydrolysis
resistant ash (HRA).
 faeces

Where nutrient refers to any nutrient such as protein, lipid or amino acid.
 Determination of Digestibility

• Advantages of this indirect method:

➢ No need to determine the total amount of food ingested or the amount of faeces
ingested.
➢ Sufficient to analyze a sample of both and determine the percentage content of
nutrient and marker.

Disadvantages of this indirect method:

➢ In some studies, the faeces are extracted from the rectum by stripping.
➢ Since digestion and absorption continue all along the digestive tract, it makes a
significant difference which part of the intestine the faeces were stripped from.
 Determination of Digestibility

• Apparent digestibility:

• True digestibility:
➢ The amount of metabolic faecal protein should be first determined and
subtracted from the protein in the faeces.
➢ Metabolic faecal protein can be determined by measuring the amount of
nitrogen excreted in the faeces when the fish is fed a protein free diet.
 True nutrient digestibility

➢ True nutrient digestibility (%)

% nutrient in feed % nutrient in faeces g MFL/100 g feed

( ----------------------- - ------------------------- - ------------------------- )
% Cr203 in feed % Cr203 in faeces % Cr203 in feed
➢ 100* --------------------------------------------------------------------------------------------
% nutrient in feed
( ------------------------- )
% Cr203 in feed

➢ Where, MFL is metabolic faecal loss or amino acids.

Factors affecting Digestibility
 Factors affecting Digestibility

• Three main factors:

➢ The ingested food and the extent to which it is susceptible to the effect of the
digestive enzymes.
➢ The activity of the digestive enzymes.
➢ The length of time the food is expected to the action of the digestive enzymes.

Each of these main factors is affected by a number of exogenous and endogenous

factors.
 Factors affecting Digestibility
• Fish species:
➢ Digestibility co-efficient may vary among the species, due both to differences in
the digestive system.
➢ A wide variation in digestibility of carbohydrates, especially starch has been
observed.
➢ Carnivorous fish digest starch to a much lesser degree than omnivorous and
herbivorous fish.

• Fish size:
➢ Enzyme activity may vary with fish age.
➢ Digestion rate or gastric emptying time in influenced by size.
 Factors affecting Digestibility
• Physiological conditions:
➢ Stressed fish, due either to excessive handling or to disease, may have a disturbed
digestibility.
➢ A long period of starvation may also affect enzyme secretion and digestibility.
➢ Starvation generally reduces the hydrolytic capacity of the intestine.

• Temperature:
➢ Fish are poikilothermic, and therefore a temperature influence on the speed at
which food is processed is to be expected.
➢ Temperature may also affect the rate of absorption of digested nutrients.
➢ Also affects the rate of passage of food through the digestive tract.
 Factors affecting Digestibility
• Water salinity:
➢ Not much is known of the effect of salinity or other factors related to water
composition on digestibility.
➢ Nutrient digestibility of Rainbow trout fell linearly with increasing water salinity
from freshwater to sea water.
• Type of food:
➢ The influence of the type of food on gastric evacuation is well known.
➢ The digestibility of food will not only affect emptying rate from the stomach but,
may also determine the time after ingestion before meal weight decrease can occur.
➢ The presence of fat in the food may delay gastric emptying, possibly by a release
of hormones.
Thanks To All
For
Patience Hearing