NUTRACEUTICALS

IN
LIVESTOCK AND POULTRY
 NUTRACEUTICALS
IN
LIVESTOCK AND POULTRY

Amitav Bhattacharyya
&
Debashis Roy

NEW INDIA PUBLISHING AGENCY

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 Contents

1. Introduction
2. Antibiotics
Poultry
Antibiotic as growth promoter in poultry feeds
Ruminants
References
3. Probiotics
Characteristics of an ideal probiotic
Poultry
Use of probiotics as growth promoter
Effect of probiotics on gut micro flora
Health and production performance
Carcass quality characteristics of the broilers
Ruminants and pigs
Bacterial DFM
Fungal DFM
Practical Considerations for DFM/Probiotics
Conclusion
References
4. Prebiotics
Characteristics of Ideal Prebiotics
Beneficial Effects of Prebiotics
Classification
Mechanism of Action of Prebiotics
Ruminants
Pigs
Poultry
Lactose
Mannan-oligosaccharide
Fructo oligosaccharides
Miscellaneous prebiotics
Partially Hydrolysed Guargum (PHGG)
Lactosucrose
Isomalto oligosaccharide
Conclusion
References
5. Enzymes
Poultry
Phytates
Non starch polysaccharides (NSP)
 Phytase
Adding Enzymes (dry) to Animal Feed
Spraying Enzymes (liquid) Directly onto Animal Feeds
References
6. Organic Acids
Poultry
Butyric acid
Acetic acid
Citric acid
Propionic acid
Lactic acid
Role of organic acid in egg production and quality parameters
Role of organic acid in immunity of poultry
Pigs
Organic acid on pig performance
Salmonella control in fattening pigs
Ruminants
References
7. Fatty Acids
Poultry
Ruminants
Biohydrogenation and Digestion of Fat
Fat Sources
Natural fats
Commercial fats
Quality Factors
Feeding Practices
Effects of Supplemental Fat on Milk Composition
Fat Analysis
Conjugated linoleic acid
CLA Chemistry
CLA Synthesis
Boosting concentrations of CLA in milk
Potential health benefits
Importance of dietary CLA in pig feeding
Conclusion
References
8. Phytobiotics
Curcuma longa (Haldi)
Terminalia arjuna
Emblica officinalis (Amla)
Azadirachta indica (Neem)
Ocimum sanctum (Tulsi)
Plant secondary metabolites
Saponins
Effects on animal growth and feed intake
Effects on the immune system
Antioxidant effects
Tannins
Definition and classification
The distribution of tannins in nature
Effect of tannins in ruminant nutrition
Practical use of tannins Treatments to protect dietary protein from ruminal degradation
Bloat prevention
Control of internal parasites
Essential oils
References
 1 Introduction

The term “nutraceutical” was coined from “nutrition” and “pharmaceutical” in 1989 by Stephen DeFelice, MD, founder and
chairman of the Foundation for Innovation in Medicine (FIM), Cranford, NJ. According to DeFelice, nutraceutical can be defined as,
“a food (or part of a food) that provides medical or health benefits, including the prevention and/or treatment of a disease”. In other
words, nutraceuticals are chemical or natural feed ingredients which may enhance health by providing a physiological benefit by the
provision of basic nutrients. Enzymes, prebiotics, probiotics, yeast and fungal extracts, predigested hydrolyzed carbohydrates and
fats, phytogenic additives, acids etc are included in nutraceuticals. However, the term nutraceutical as commonly used in marketing
has no regulatory definition.
A nutraceutical is a product isolated or purified from foods that is generally sold in medicinal forms not usually associated with food.
A nutraceutical is demonstrated to have physiological benefit or provide protection against chronic disease. On the other hand,
“functional feed” is similar in appearance to a conventional food that is consumed as a part of usual diet, and is demonstrated to
have physiological benefits to reduce the risk of chronic disease beyond basic nutritional function, i.e. they contain bioactive
compound. The different types of nutraceuticals have been classified in Table 1.1. However, only few of them have been studied in
detail pertaining to their specific mode of action in different species, synergistic action etc. Studies on nutraceuticals have gained
importance after the ban on sub-therapeutic use of antibiotics in several countries in the last century. It is expected that the trend will
continue in the present century too along with a search for newer nutraceuticals for optimum production in livestock and poultry.
Table 1.1. Different types of nutraceuticals used in livestock, poultry and pig

Nutraceutical Examples Expected Actions Research Support/Status

Type
Antioxidants Vitamin E & Selenium; Minimize A significant number of
Vitamin C; Grape seed oxidative studies have been published
extract; lipoic acid etc. damage to addressing various aspects
reactive oxygen of antioxidant effects. Study
species results have been mixed, but
generated during most suggested of some
intense exercise positive effect.
or inflammatory
conditions.
Antimicrobials Antibiotics; Provide therapy Highly regulated by FDA
Anthelmintics etc. or prevention of and must be used
specific pathogen accordingly.
associated
diseases and act
as growth
promoter.
Herbal additives Wide range of whole Anti- Lot of studies have been
plants or Extracts. methanogenic conducted recently on their
antiinflammatory; effect as rumen fermentation
metabolic modulator.
effects.
Enzymes Phytase; Cellulase; Improve Fiber digestibility is
Hemicellulase digestive reported to be increased
capacity of with enzymes.
specific
compounds,
nutrients within
consumed diet
Ergogenic Chromium; Creatine; Improve muscle A limited number of studies
Agents Betaine (TMG); mass and have been published on
Dimethyl-glycine function. various agents in case of
(DMG) Increase muscle pigs. Most have shown no
capacity for effect of supplement
aerobic compared to controls.
metabolism and
facilitate
recovery
Oligosaccharides Fructooligosaccharides; Alter intestinal Limited studies published.
mannan- microbial Potential for use in pigs and
oligosaccharides etc. populations by calves. But needs further
either stimulating study.
growth of
beneficial
bacteria or
inhibiting
pathogens
Organic Chelates; Improve Studies varied in type of
Minerals Proteinates;Complexes; bioavailability of organic source versus
Polysaccharides mineral elements inorganic source. Limited
from the diet effects were seen with
organic mineral
supplementation and mostly
with younger animals.
Omega-3 Fatty EPA, DHA, Flaxseed, Anti- Studies showed alteration in
Acids Fishoils inflammatory either plasma or membrane
properties, fatty acids profile based on
counter source. Most studies
regulatory showed anti-inflammatory
 metabolically to effects with n-3 fatty acids.
n-6 fatty acid Clinical studies suggest
derived reduced disease skin
eicosanoids allergies with n-3
compounds supplementation.
Organic acids Acetic acid, Propionic It reduces Studies on various organic
acid, Butyric acid, intracellular pH acids conducted in
citric acid and thereby ruminants, pigs and poultry
reduces enzyme
activity leading
to death of
bacteria
Probiotics Various lactic acid Alter ruminal Mixed results reported and
producing bacteria fermentation suggested beneficial effect
(live cultures). pattern. Alter as rumen modulator.
intestinal
microbial
populations by
providing live
beneficial
bacteria that
inhibit pathogen
growth.
Yeast and Live or culture extracts Improve fiber Most studies showed some
Yeastextracts Saccharomyces digestibility and improvement in nutrient
cerevisiea dietary nutrient availability. Some studies
availability; have documented improved
Alter intestinal performance.
microbial flora.
 2 Antibiotics

Feed accounts for a major portion of farm expenses in livestock and poultry production. Thus, to obtain maximum profitability, it
must be ensured that the feed should not only be well balanced nutritionally but economical too. Rising prices of feeds certainly have
reduced the profitable nature of broiler farming. For better utilization of feed and to improve feed efficiency, antibiotics were used at
sub-therapeutic levels in the animal ration.
Development of antibacterials progressed very quickly and as a result, certain new antibacterial substances were discovered and
developed for use as growth promoting agents in livestock and poultry. The antibiotics act by reducing nutrient destruction in the
intestine, increasing microbial synthesis of certain vitamins and amino acids, reducing the rate of passage of nutrients in the
gastrointestinal tract and reducing the thickness of the intestinal wall. This thinner wall and reduced rate of passage have been
suggested as factors favoring efficient absorption of nutrients. It is also reported that the antibiotics also inhibit the bacterial
production of ammonia and other harmful nitrogenous compounds such as trimethylamine that reduce the growth of chickens.

Poultry
Antibiotics have been used at sub-therapeutic levels for promoting the growth and immunity of birds. The advantages of using
antibiotics as feed supplements in terms of growth stimulation and improvement of feed efficiency have been well documented
(Ensminger et al., 1990; Peterson et al., 1991).

Antibiotic as growth promoter in poultry feeds

George et al. (1982) noted that birds fed virginiamycin had significantly less mortality and lower intestinal lesion scores compared to
non-medicated birds when experimentally infected with necrotic enteritis. Abou et al. (1983) reported that virginiamycin
administered to the experimentally infected group for 8 weeks in the feed of broilers at a concentration of 25 g/ton had no beneficial
effect compared to an infected control group not receiving the antibiotic.
Body weight and bone ash at 21-days of age were improved by the addition of virginiamycin in the diet of broilers. Also, the amount
of phosphorus required to produce a gram of body weight was decreased by the addition of virginiamycin in the diet (Buresh et al.,
1985).
Bartov et al. (1992) reported that virginiamycin had no significant effect on food intake or weight gain, but significantly improved
food efficiency up to 28 days of age in broiler chicks. Further, virginiamycin significantly decreased fat excretion and improved fat
relative retention. Similarly, Al-Batshan et al. (1992) noted that virginiamycin improved body weight and feed efficiency from 1 to 29
days of age in turkey poults, irrespective of type of litter or disease condition such as stunting syndrome.
Proudfoot et al. (1990) noted that virginiamycin supplementation had no significant effect on mortality or feed conversion ratios in
male chicken broilers, regardless of the mode of administration. However, body weights at 21 days of age but not at 42 days of age
were significantly heavier for broilers receiving virginiamycin via the drinking water. The inclusion of virginiamycin in the feed failed
to improve body weights at either 21 or 42 days of age. Miles et al. (1984) reported that diet supplemented with virginiamycin
resulted in an increase in xanthophyll utilization and feed efficiency was significantly improved by the supplementation of 5 and 10
ppm of virginiamycin in the broiler diet. Salmon et al. (1990) noted that the combination of virginiamycin and monensin resulted in
superior overall feed efficiency to that obtained with either additive alone in turkey broilers.
Tokosova et al. (1990) studied the effect of cyadox and virginiamycin on Marek’s disease in chickens. The weight gains on the 56th
day and dressing percentage were highest, in comparison to the control group, in the virginiamycin-treated group. Decrease in heart
weight occurred in the cyadox treated group where as increase in liver and heart weight and intestine shortening occurred in the
virginiamycin treated group as compared to the control group. Woodward et al. (1988) noted that the body weights were higher at
higher energy levels with addition of virginiamycin to the diets. Further, yield was increased from 63.3 to 64.0% by dietary
virginiamycin.
Henry et al. (1986) reported that the virginiamycin decreased relative intestinal tract weight and increased kidney and bone
manganese indicating that virginiamycin increased absorption of manganese.

Ruminants
The ruminant animal does not efficiently convert feedstuffs into meat or milk. As a result, several strategies have been used to
improve ruminant feed efficiency. One of the techniques to improve the efficiency of the fermentation has included the addition of
antimicrobial compounds in the diet to alter the ruminal microbial ecosystem. Ionophores, antibiotics e.g. monensin, lasalocid,
laidlomycin, salinomycin, narasin etc. and conventional antibiotics e.g. chlortetracycline, oxytetracycline, bacitracin, tylosin etc. are
antimicrobial compounds that are commonly fed to ruminant animals to improve feed efficiency.
Ionophores are antibiotic class that alters rumen fermentation characteristics. Ionophores cause cattle to grow more efficiently
(Russell and Strobel, 1989) but were originally used to control intestinal parasites (coccidiostat) in poultry (Bergen and Bates, 1984).
Monensin has been marketed for cattle as a methane inhibitor and propionate (the most efficiently utilized gluconeogenic VFA)
enhancer (Dinius et al., 1976; Richardson et al., 1976; Russell and Strobel, 1989). Additional benefits of monensin usage include a
reduction of dietary protein deamination, resulting in less ammonia urinary excretion (Russell and Strobel, 1989) and a decrease in
lactic acid production (Dennis et al., 1981) which results in a reduction in ruminal acidosis (Russell and Strobel, 1989) and liver
abscesses (Nagaraja and Chengappa, 1998). The increases in energy availability and nitrogen retention improve the efficiency of
feed utilization by the ruminant animal, and thus improve animal productivity and production profitability (Potter et al., 1976; Russell
and Strobel, 1989). Monensin treatment reduces morbidity and mortality among feedlot animals by reducing the incidence of acute
and sub-acute ruminal acidosis, bloat, and bovine emphysema (Galyean and Owens, 1988). Dietary carbohydrates are rapidly
fermented in the rumen which can result in an accumulation of lactic acid, resulting in a lowered ruminal pH, ruminal dysfunction and
acidosis (Nagaraja et al., 1982; Burrin and Britton, 1986; Russell and Rychlik, 2001). Ruminal acidosis is associated with reduced
feed intake, lowered feed efficiency and cyclic feeding, as well as the death of some animals. Monensin reduces acidosis by directly
inhibiting the lactate-producing bacteria (e.g., Streptococcus bovis, lactobacilli) (Dennis et al., 1981).
All improvements in animal productivity caused by ionophore treatment represent a secondary effect caused by the disruption of
normal bacterial membrane physiology (Bergen and Bates, 1984). Ionophores are moderate molecular weight compounds (~700
MW) that are mobile ion carriers (Pressman, 1976). Because ionophores are highly lipophilic, they rapidly dissolve into bacterial cell
membranes (Pressman, 1976). Ionophores bind ions, shield the ionic charges and translocate ions across the bacterial membrane,
disrupting crucial ion gradients (Pressman, 1976). Because ionophores are lipophilic compounds that exert their effects at the
membrane level, they are most effective against gram-positive bacteria. The peptidoglycan layer that surrounds gram-positive
bacteria is porous, and allows small molecules to pass through, reaching the cytoplasmic membrane where the lipophilic ionophore
rapidly dissolves into the membrane. Conversely, gram-negative bacteria, are separated from the environment and antimicrobial
agents by a lipopolysaccharide layer, outer membrane and periplasmic space. Monensin is bound by both grampositive and gram-
negative bacteria (Chow et al., 1994).
Ionophores are potent antimicrobial compounds that improve production efficiency and health in cattle by altering the composition of
the ruminal microbial ecosystem, thereby reducing the incidence of illnesses related to the ruminal fermentation (e.g., bloat, bovine
emphysema). However, like all other antimicrobial compounds, concerns have been raised about the development of antimicrobial
resistance and the potential for the transfer of cross resistance to antibiotics used in human medicine.
Conventional antibiotics have been used in animal feed for about 50 years ever since the discovery not only as an anti-microbial
agent, but also as a growth-promoting agent and improvement in performance. Tetracyclines, penicillin, streptomycin and bactrican
soon began to be common additives in feed for livestock. Currently, the following antibiotics are used in livestock: chlortetracycline,
procaine penicillin, oxytetracycline, tylosin, bacitracin, neomycin sulfate, streptomycin, erythromycin, linomycin, oleandomycin,
virginiamycin and bambermycins. In addition to these antibiotics, which are of microbial origin, there are other chemically
synthesized antimicrobial agents that are also sometimes used in animal feeds. These include three major classes of compounds:
arsenical, nito-furan and sulfa compounds. Arsenical compounds include arsanilic acid, 3-nitro-4-hydroxy phenylarsonic acid and
sodium arsanilate; nitro-furan compounds include furazolidone and nitro-furazone; sulpha compounds include sulfamethazine,
sulfathiazole, and sulfaquinoxaline. Antibiotics are used regularly in animal feed in many countries at a rate of 2 to 50 grams per ton
for improved performance in the animals (McDonald et al., 1997). The reasons include a more efficient conversion of feed to animal
products, an increased growth rate and a lower morbidity/mortality rate in general. The levels of antibiotics are often increased to
50-200 grams/ton or more when specific diseases are being targeted as when the spread of a particular disease is rampant. The
levels are also increased in times of stress. This increased amount is often decreased when the threat of a disease is gone.
After animals have been fed antibiotics over a period of time, they retain the strains of bacteria which are resistant to antibiotics
(FDA, 1995). These bacteria proliferate in the animal. Through interaction, the resistant bacteria are transmitted to the other
animals, thus forming a colonization of antibiotic resistant bacteria. The bacteria flourish in the intestinal flora of the animal, as well
as, in the muscle. As a result, the feces of the animal often contain the resistant bacteria. Transfer of the bacteria from animal to
human is possible through many practices. The primary exposure of humans to resistant bacteria occurs in farms and
slaughterhouses.
The possibility of developing resistant population of bacteria and side effects of using antibiotics as growth promoters in farm animals
have been of immense concern, principally with regard to the loss of efficacy of antibiotic as growth stimulant and for controlling an
outbreak of bacterial disease (Hinton, 1988). Some early studies showed that continuous feeding of antibiotics to chickens resulted in
a decrease growth response. The possible emergence of antibiotic-resistant strains of bacteria on the premises where growing birds
are fed on low concentration of antibiotics has been reported (Narayanankutty et al., 1992). Following the discovery of resistance
transfer factor, there has been growing concern about public health risk resulting from antibiotic resistance, carcinogenic responses
and other side effects of residues in food. The most important potential route by which humans become infected with resistant
bacteria is through meat, milk and eggs (Hinton, 1988). As poultry meat is one of the most important sources of animal protein
throughout the world, even a low incidence of cross infection could be important. The possibility of developing resistant bacteria
besides other side effects when antibiotics are used as growth promoters have led to the recent EU ban on the use of most
antibiotics on farm animals as growth promoters. This ought to have serious consequences in growth performance of poultry.
Therefore, an intensive search for alternative feed additives started in the last decade.

References
Abou Youssef, M. H., Di Cuollo, C. J., Free, S. M. and Scott, G. C. 1983. The influence of a feed additive level of virginiamycin on
the course of an experimentally induced Salmonella typhimurium infection in broilers. Poultry Science, 62: 30-37.
Al-Batshan, H. A., Sell, J. L., Piquer, J., Mallarino, E., Soto-Salanova, M.F. and Angel, C.R. 1992. Responses of turkey poults to
virginiamycin as influenced by litter condition and experimentally induced stunting syndrome. Poultry Science, 71: 894-904.
Bartov, I. 1992. Lack of effect of dietary energy-to-protein ratio and energy concentration on the response of broiler chickens to
virginiamycin. British Poultry Science, 33: 381-391.
Bergen, W.G., and Bates, D.B. 1984. Ionophores: Their effect on production efficiency and mode of action. Journal of Animal
Science, 58: 1465-1483.
Buresh, R. E., Miles, R. D. and Harms, R. H. 1985. Influence of virginiamycin on phosphorus utilization by broiler chicks. Poultry
Science, 64: 757-758.
Burrin, D.G. and Britton, R.A. 1986. Response to monensin in cattle during subacute acidosis in cattle. Journal of Animal Science,
63: 888-893.
Chow, J.M., Van Kessel, J.A.S. and Russell, J.B. 1994. Binding of radiolabeled monensin and lasalocid to ruminal microorganisms
and feed. Journal of Animal Science, 72: 1630-1635.
Dennis, S.M., Nagaraja, T.G. and Bartley, E.E. 1981. Effects of lasalocid or monensin on lactate-producing or -using rumen
bacteria. Journal of Animal Science, 52: 418-426.
Dinius, D.A., Simpson, M.E. and Marsh, P.B. 1976. Effect of monensin fed with forage on digestion and the ruminal ecosystem of
steers. Journal of Animal Science, 42: 229-234.
Ensminger, M. E., Oldfield, J. E., and Heinemann, W. 1990. Feeds and Nutrition. Ensminger Publishing Company, USA.
FDA. 1995. The Rise of Antibiotic-Resistant Infections. Consumer, 29. http://www.fda.gov/fdac/features/795_antibiotic.html
Galyean, M.L., and Owens, F.N. 1988. Effects of monensin on growth, reproduction, and lactation in ruminants. In: ISI Atlas of
Science: Animal and Plant Science, ed. ISI Press, Philadelphia, PA. p. 71-75.
George, B. A., Quarles, C. L. and Fagerberg, D. J. 1982. Virginiamycin effects on controlling necrotic enteritis infection in chickens.
Poultry Science, 61: 447-450.
Henry, P. R., Ammerman, C. B. and Miles, R. D. 1986. Influence of virginiamycin and dietary manganese on performance,
manganese utilization, and intestinal tract weight of broilers. Poultry Science, 65: 321-324.
Hinton, M. H. 1988. Antibiotics, poultry production and public health. World’s Poultry Science Journal, 44: 67-69.
McDonald, L. C., Matthew J. K., Fred C. T. and William R. J. 1997. Vancomycin-Resistant Enterococci Outside the Health-Care
Setting: Prevalence, Sources, and Public Health Implications. Centers of Disease Control and Prevention, Alanta,
Georgia, 3. wysiwyg://67/http://www.cdc.gov/ncidod/EID/vol3no3/mcdonald.htm
Miles, R. D., Janky, D. M. and Harms, R. H. 1984. Virginiamycin and broiler performance. Poultry Science, 63: 1218-1221.
Nagaraja, T.G. and Chengappa, M.M. 1998. Liver abscesses in feedlot cattle: A review. Journal of Animal Science, 76: 287-298.
Nagaraja, T.G., Avery, T.B., Bartley, E.E., Roof, S.K. and Dayton, A.D. 1982. Effect of lasalocid, monensin or thiopeptin on lactic
acidosis in cattle. Journal of Animal Science, 54: 649-658.
Narayanankutty, K., Ramakrishnan, A. and Viswannath, A. 1992. Efficacy of virginiamycin as growth promoter in commercial
 broiler chicks. Journal of Veterinary and Animal Sciences, 23: 96-97.
Peterson, R. A., Carpenter, G. H. and Jones, W. T. 1991. Effect of tetracycline hydrochloride and oxytetracycline hydrochloride
given via drinking water on early mortality of broiler chicks from twenty- eight-week-old dams. Poultry Science, 70: 1040-
1042.
Potter, E.L., Raun, A.P., Cooley, C.O., Rathmacher, R.P. and Richardson, L.F. 1976. Effect of monensin on carcass
characteristics, carcass composition and efficiency of converting feed to carcass. Journal of Animal Science, 43: 678-683.
Pressman. 1976. Biological applications of ionophores. Annual Review of Biochemistry, 45: 501-530.
Proudfoot, F. G., Jackson, E. D. and Hulan, H. W. 1990. The response of male chicken broilers to the dietary addition of
virginiamycin. Poultry Science, 69: 1713-1717.
Richardson, L.F., Raun, A.P., Potter, E.L., Cooley, C.O. and Rathmacher, R.P. 1976. Effect of monensin on rumen fermentation in
vitro and in vivo. Journal of Animal Science, 43: 657- 664.
Russell, J.B. and Rychlik, J.L. 2001. Factors that alter rumen microbial ecology. Science, 292: 1119-1122.
Russell, J.B. and Strobel, H.J. 1989. Effect of ionophores on ruminal fermentation. Applied and Environmental Microbiology, 55:
1-6.
Salmon, R. E. and Stevens, V. I. 1990. Virginiamycin and monensin, alone or in combination, in turkey broiler diets. Poultry
Science, 69: 1016-1019.
Tokosova, M. 1990. Observations of the effects of cyadox and virginiamycin on the course of Marek’s disease in chickens.
Veterinarni Medicina, 35: 105-112.
Woodward, S. A., Harms, R. H., Miles, R. D., Janky, D. M. and Ruiz, N. 1988. Influence of Virginiamycin on yield of broilers fed
four levels of energy. Poultry Science, 67: 1222-1224.
 3 Probiotics

The increasing use of antibiotics in livestock and poultry led to development of antibiotic resistant micro organisms and antibiotic
residues in livestock products besides other side effects. These caused alarm bells ringing all over the world and by the end of June,
1999 majority of antibiotics as growth promoters in monogastric diets had been banned within the EU. The ban on use of antibiotic at
sub therapeutic level in Europe and the potential for a ban in the United States have made scientists to seriously think of alternatives
to antibiotics. In fact, the use of probiotics for farm animals was already stimulated by the findings of the Schwann Committee in
1969 which recommended that antibiotics in animal feeds should be restricted. However, the concept of probiotics gained
momentum in later part of the last century. In recent years, with the public disapproval of using antibiotics in livestock and poultry
production due to their residual effects in their products, there has been a greater emphasis on the use of probiotics in poultry.
‘Probiotic’ is a word derived from Greek word ‘pro’ and ‘biotic’ meaning ‘for life’. Probiotics can be described as organisms and
substances which contribute to intestinal microbial balance (Sperti, 1971). Fuller (1989) revised the definition as a live microbial feed
supplement, which beneficially affects the host animal by improving the intestinal microbial balance.
The term probiotic was coined by Parker (1974) to describe organisms and substances, which contribute to intestinal microbial
balance. However, the use of probiotic was first suggested by Elias Metchnikoff (1907) as he realized the role of probiotics in
combating various disorders of gastrointestinal tract. He explained that the detrimental microbes in the intestinal tract excreted
substances that were harmful to the host, through the constant infusion of ‘friendly’ organisms in the diet. Colonization of the
gastrointestinal tract by disease causing (pathogenic) unfriendly organism was prevented and thus health and life expectancy
improved. Thus the concept of microbial inoculation, based on the principle of ‘competitive exclusion’ was established.
Marriott and Davidson (1923) reported that young infant fed similar quantities of fresh cow milk or milk to which lactic acid was
added had higher mean daily weight gains. Since then, the different types of probiotics have been tested like lactic acid producing
bacteria, live yeast culture etc. In fact, nearly all the probiotics available in the market today contain Saccharomyces cereviscae ,
bacteria or their combination.
Probiotic may be classified into two major types: viable microbial cultures viz. Lactobacilli, Streptococci, Bacillus etc and microbial
fermentation products. Basically a delicate balance exists between beneficial and healthy bacteria in a healthy bird and an imbalance
leads to dysbacteriosis. Optimum growth of both beneficial and pathogenic micro organism is controlled by pH of intestine. This
imbalance may be caused due to various factors like contaminated feed and water, environmental stress, overcrowding, prolonged
use of antibiotics, infection etc. Under these changed circumstances, the putrefactive bacteria predominate and impair the normal
gut function. Among all the species of bacteria that help in maintaining the normal gut function, Lactobacilli species is most affected
under the changed circumstances and ultimately fail to maintain proper pH (4.5).
Several mechanisms have been postulated to explain the mode of action of probiotics. They are as follows:
Probiotics cause beneficial changes in the gut flora with reduction in population of E.coli. Lactobacilli, the most
common microbe of probiotic compete with coliforms and pathogenic microorganisms for the site of adherence on the
intestinal surface.
Probiotics like lactobacillus increase lactic acid production with subsequent change in intestinal pH. Lactic acid
release in the intestine reduces the pH up to 4.5, which is lethal for pathogens but allow the growth of some strains of
Lactobacilli.
Probiotics produce antibiotic type substances. Live yeast culture is capable of releasing phytase for phosphorous
digestion, lipase for fat digestion and endotryptase for protein digestion. It triggers the growth of cellulolytic bacteria
resulting into digestion of fibre portion of feed. The yeast also facilitates the growth of lactobacillus. Yeast ferments
the carbohydrate. The fermentation products inactivate the intestinal toxins that reduce the appetite of poultry. Yeasts
 are also a rich source of vitamin B complex.
Probiotics reduce the release of toxins probably with suppression of E.coli.

Characteristics of an ideal probiotic

It should be capable to survive and metabolize in the gut environment i.e. it should be resistant to low pH.
It should multiply faster in the digestive tract.
It should be capable of exerting a beneficial effect on the host animal i.e. elicit growth and production.
It should not be pathogenic or toxic to the host.
The adhesive capacity of the probiotics should be firm and faster.

Poultry
Jin et al. (1997) concluded that the mode of action of probiotics in poultry includes (1) maintaining normal intestinal micro flora by
competitive exclusion and antagonism. (2) Altering metabolism by increasing digestive enzyme activity and decreasing bacterial
enzyme activity.

Use of probiotics as growth promoter

Fuller (1995) reported beneficial effects on growth rate, feed conversion efficiency and resistance to disease in broilers fed
probiotics. Since then, different types of probiotics have been used singly or in combination to improve growth and FCR in different
species of poultry.

Effect of probiotics on gut micro flora

Probiotics help in maintaining or restoring a stable micro flora in the gut of chickens that has been affected by pathogens and growth
depressing bacteria (Barrow, 1992). Thus, use of probiotics based on specially selected bacteria have been suggested to normalize
the intestinal micro flora and increase resistance to physical and physiological stress in intensively reared chickens (Tibiletti, 1993). It
has been seen that microbial combination of different strains of Lactobacilli, Streptococci and Sacchromyces cerevisae from
indigenous sources fed up to the grower phase were well established in the gastro intestinal tract of birds, eliciting beneficial effect
throughout the laying period (Sharma et al., 2001).

Health and production performance

It has been noted that dietary supplementation of probiotics in broilers resulted in a significant reduction on the total serum
cholesterol concentration and crop Escherichia coli count (Panda et al., 2001). Further, health and production traits of broilers are
positively influenced and improved by feeding probiotics. Addition of probiotic to broiler diets reduce mortality rate due to ascites
(Kahraman et al., 1997).
Use of probiotics in layer diets result in better hen body weight, egg production, egg weight, egg mass, egg component percentages,
egg shell thickness, egg shell weight, feed intake, feed conversion, livability, decrease in meat & blood spots and decreased egg yolk
and serum cholesterol (Huthail, 1996; Yakout et al., 2004 and Kurtoglu et al., 2004).
Use of probiotic in chicken diet results in reduced abdominal fat content, improved fatty acid digestibility, increased mineral utilization
including organic phosphorus utilization, increased tibial ash and enhanced feed efficiency.
Use of probiotic in poultry diets may elicit the immunocompetence traits in poultry (Panda et al., 2008 & Dalloul et al., 2003).

Carcass quality characteristics of the broilers

Supplementation of probiotics in broiler diets may result in significant improvement in gain, feathering, dressing and edible meat yields
without any undesirable change in the chemical composition and organoleptic quality of broiler meat (Krueger et al., 1990, Yalcin et
al., 1993 & Banday and Risam, 2001).

Ruminants and pigs

Before birth, developing animals are sterile in the womb of their mothers. Upon birth, the digestive tracts of all animals are naturally
colonized by a variety of microorganisms (Savage, 1987). Under healthy and non-stressful conditions, ”beneficial” microflora
colonizes the rumen and lower gut in a symbiotic relationship with the host. Beneficial rumen and gut microorganisms supply
nutrients to the host, aid in digestion of dietary nutrients and compete with potential pathogens. In contrast, when young animals are
removed and raised under sterile conditions, microorganisms from the environment are prevented from colonizing their digestive
tracts. These animals often have increased nutritional needs (e.g., requiring more vitamin K in the diet) and abnormal immune
responses. Sterile animals also are more susceptible to bacterial infections, presumably due to rapid establishment of pathogens.
The original concept of administering direct fed microbial to animals was to feed large amounts of “beneficial” microbes to livestock
when they were “stressed.” This practice would prevent the establishment of pathogenic microorganisms and could help re-establish
normal gut microflora. This practice was termed “probiotic” or “for life.” The term “probiotic” implied a curative nature of these
products that would require government approval in order to make legal product claims (e.g., decreased mortality, fewer sick days,
and increased production).
One of the most common hypotheses that may explain how probiotics improve animal performance suggests that the addition of
beneficial bacteria exclude the establishment of pathogens (competitive exclusion). Production of antimicrobial end products such as
acids and antibiotics are also commonly discussed.

Bacterial DFM
Probiotics are live micro organisms fed directly to the animals. Thus, they are also known as direct fed microbials (DFM). In
general, most would agree that DFM based on bacteria must be “live.” Thus, they must survive processing, storage and the gut
environment. However, future research may prove that end products such as bacteriocins (narrow spectrum antimicrobial
substances) and not the actual organism itself may be beneficial. A list of some common bacteria that have potential as DFM
additives is shown in Table 3.1.
Lactobacillus acidophilus (and other Lactobacillus species), L. casei, Enterococcus diacetylactis, and Bacillus subtilis are
commonly used as DFM products for ruminants. These organisms appear to have little effect on ruminal fermentation (Ware et. al.,
1988) and the site of action from these organisms appears to be in the lower gut but solid and repeatable data is lacking. Initial
research with these organisms in ruminants was first centered on “stressed” animals with the general assumption that feeding
beneficial organisms would decrease or prevent intestinal establishment of pathogenic microorganisms (Vandevoorde et al., 1991).
In addition, it was hypothesized that massive doses of beneficial organisms would re-colonize a “stressed” intestinal environment and
return gut function to normal more quickly. In ruminants, much of this research involved feeding lactobacillus-based DFM to young
calves fed milk, calves being weaned or shipped cattle (Jenny et al., 1991; Hutcheson et al., 1980) because these conditions were
often classified as times of high stress. Calves fed L. acidophilus have been reported to have reduced incidence of diarrhea
(Beecham et al., 1977) and reduced counts of intestinal coliform bacteria (Bruce et al., 1979). Data summarizing more than 30 trials
with incoming feedlot cattle showed an advantage of 10.7 and 5.4% in average daily gain and feed efficiency, respectively, for cattle
fed a DFM (Pioneer Hi-Bred International, 1988). Only a few studies have documented positive effects of feeding bacterial DFM to
lactating dairy cows. High producing cows in early lactation would be the best candidates for such products because these cows are
in negative energy balance and have diets that contain highly fermentable carbohydrates that sometimes lead to acidosis. Jaquette et
al. (1988) and Ware et al. (1988) reported increased milk production from cows fed L. acidophilus (1 x 109 colony-forming units
per head per day). It has also been reported improvements in milk production when cows were fed a DFM containing yeast and 2
strains of bacteria. Supplementation of lactobacilli may be useful in the close-up dry period of lactation when intake is depressed and
animals are stressed. However, there is limited data to support this use.
To some extent, the practice of using DFM on farm is already being used on many dairies. Specifically, producers and veterinarians
have been inoculating sick ruminants with rumen fluid from healthy animals in hopes of stimulating normal rumen function for
improving dry matter intakes has been practiced for decades. Several attempts have been made to use bacteria to alter rumen
metabolism but only a few have been successful on a practical scale.
The detoxification of the 3-hydroxy-4(1H)-pyridone (DHP) by Synergistes jonesii, isolated from Hawaiian cattle, is probably one of
the most cited successes of manipulating ruminal fermentation with bacteria. The tropical forage Leucaena leucocephala (Subabul
in Hindi) contains mimosine, a non-protein amino acid. When consumed by ruminants in Australia and some parts of India, DHP
causes goitrogenic effects. Jones and Megarrity (1986) showed that rumen microbes, from cattle in Hawaii, were able to detoxify
DHP. The specific organism responsible for detoxification, S. jonesii (Allison et al., 1990), was inoculated and established itself in
the rumen of Australian cattle thus conferring protection from DHP toxicity. Another problem in feeding ruminants, identified in
Australia, is monofluroacetate. This compound is found in some Australian plants and can be toxic to ruminants at doses of about 0.3
mg/kg of body weight. Gregg et al. (1998) reported that they successfully inserted the gene encoding for fluoroacetate dehalogenase
into several strains of Butyrivibrio fibrisolvens and when sheep were inoculated with the altered microbes, they showed reduced
toxicological symptoms. However, use of the genetically modified rumen bacteria in the field is not currently approved.
Megasphaera elsdenii (ME) is the major lactate-utilizing organism in the rumen of adapted cattle fed high grain diets. However,
when cattle are abruptly shifted from a high-forage to high-concentrate diet, the numbers of ME are often insufficient to prevent
lactic acidosis. We have shown that during a challenge with highly fermentable carbohydrates, addition of Megasphaera elsdenii
B159 prevented an accumulation of lactic acid and shifted ruminal fermentation away from acetate and propionate towards butyrate
and valerate (Kung and Hession, 1995). Addition of ME has also experimentally prevented acidosis in steers (Robinson et al., 1992).
Development of this organism for feedlot cattle, and perhaps high producing dairy cows, should be continued with emphasis on
optimizing dose and timing of administration. Success with such an organism could allow feedlot producers to decrease the time it
takes to adapt cattle to a high concentrate diet. It could also be useful by reducing chronic acidosis in lactating cows.
Some Propionibacteria are naturally found in high numbers in the rumen of animals fed forage and medium concentrate diets.
These organisms convert lactate and glucose to acetate and propionate. Propionibacteria may be beneficial if inoculated into the
rumen (Kung et al., 1991) because higher concentrations of ruminal propionate would be absorbed into the blood and converted to
glucose by the liver of the host animal. Although Propionibacteria can metabolize lactic acid, they are probably too slow growing
and acid intolerant to prevent a challenge that would lead to acidosis. A commercially available product based on a strain of
Propionibacteria that naturally occurs in the rumen has been claimed to reduce the chance of nitrate toxicity but definitive data is
lacking. Recently, Swinney-Flyod et al. (1999) reported that feedlot cattle fed a diet containing Propionibacteria, strain P-63 (1 ×
109 cfu/head/day) and L. acidophilus, strain 5345, (1 × 108cfu/head/day) had better feed efficiencies during adaptation to a high
concentrate diet and during a 120-d feeding period. Similarly, Huck et al. (1999) reported that cattle fed L. acidophilus (5 × 108
cfu/head/day) strain BG2F04, and P. freudenrechii (1 × 109 cfu/head/day) had better feed efficiencies than those fed a control
diet. More research in these areas is warranted.

Fungal DFM
A variety of mechanisms have been put forth to explain changes in ruminal fermentations and improvements in performance when
ruminants are fed fungal-based DFM. For example, yeast may have a buffering effect in the rumen by mediating the sharp drops in
rumen pH, which follows feeding. Martin and Streeter (1995) suggested that fungal cultures improve the use of lactate by the
ruminal organism Selenomonas ruminantium by providing a source of dicarboxcylic acids (e.g., malic acid) and other growth
factors. Thus, yeast may help to buffer excess lactic acid production when ruminants are fed high concentrate diets. The effects on
buffering are subtle; as added yeast cannot prevent lactic acidosis if the rumen is challenged with a diet rich in fermentable
carbohydrates (Aslan et al., 1995; Dawson and Hopkins, 1991). However, a higher pH may be one reason for the finding of
increased numbers of rumen cellulolytic bacteria and improvements in fiber digestion with fungal cultures (Arambel et al., 1987).
Newbold et al. (1995b) reported that the stimulation of rumen bacteria by Saccharomyces cerevisiae different with specific strains.
Some fungal extracts have been suggested to contain esterase enzymes that may improve fiber digestion (Varel et al., 1993). Yeast
may also stimulate rumen fermentation by scavenging excess oxygen from the rumen (Newbold et al., 1996). They have also been
shown to stimulate acetogenic bacteria in the presence of methanogens (Chaucheryas et al., 1995). The effect of fungal cultures on
ruminal VFA has been inconsistent. Newbold (1995a) summarized the literature and reported that fungal extracts had no effect or
tended to increase the rumen acetate:propionate ratios while active yeast either had no effect or decreased the acetate:propionate
ratio. Arizona researchers reported that feeding AO to cows in hot environments decreased rectal temperatures in some but not all
studies (Huber et al., 1994). There is no direct evidence that yeast or fungal extracts affect digestion or metabolism in the lower gut.
However, the potential for such effects have not been well studied.
The need for high numbers of live fungal organisms in fungal DFM additives has been the subject of many debates. Some products
guarantee live yeast cells (e.g., 1 x 109 cfu per g) and are fed at low inclusion rates (only 10-20 grams per day) but other products
suggest that live organisms are not required for beneficial effects because end products present in the additives are the “active”
ingredients. Newbold et al. (1991) reported that autoclaving, but not irradiation, decreased the ability of an AO extract to stimulate
rumen bacterial growth and activity. Dawson et al. (1990) reported that the stimulatory effect of yeast on numbers of rumen
cellulolytic bacteria was negated when yeasts were autoclaved. Martin and Nibs (1992) reported that unpublished data from their lab
showed enhanced uptake of D-lactate by S. ruminantiumwas enhanced by a filtrate from AO but not from SC. Although there
have been implications that suggests yeasts were able to grow in continuous rumen cultures (Dawson et al., 1990) others have
observed that live yeasts are essentially washed out of ruminal fermentations. We reported that Saccharomyces cerevisiae did not
multiply in sterile ruminal fluid, but they did survive and were metabolically active (Kung et al., 1996).
In contrast to research with bacterial DFM, there is much data on the effect of feeding fungal DFM to lactating cows. In a review
of 32 lactation comparisons conducted with yeast between 1986 and 1997, these supplements increased milk production on average
by more than 1.13 kg (2.49 lb.) per day with the response being greater for cows in early lactation. Response appeared to be
consistent over the years. In a summary of 26 comparisons where fungal extracts (from Aspergillus oryzae) were fed to lactating
ruminants, we found an average increase in milk production of only 0.45 kg (1.01 lb.) of milk per day. Unexplainably, since 1991,
milk production responses from fungal extracts (AO) have been relatively poor. Fungal cultures have also been fed to calves, sheep,
and steers but applications with these species have been less researched than with lactating cows. For example, Beharka et al.
(1991) reported that young calves fed an AO fermentation extract were weaned one wk earlier than untreated calves and that
supplementation increased the numbers of rumen bacteria and VFA concentrations.
From a practical point, fungal additives appear to be more useful when fed to cows in early lactation that are consuming high
quantities of grain,

Practical Considerations for DFM/Probiotics

Direct-fed microbial products are available in a variety of forms including powders, pastes, boluses, and capsules. In some
applications, DFM may be mixed with feed or administered in the drinking water. However, use of DFM in the latter manner must
be managed closely since interactions with chlorine, water temperature, minerals, flow rate, and antibiotics can affect the viability of
many organisms. Non-hydroscopic whey is often used as a carrier for bacterial DFM and is a good medium to initiate growth.
Bacterial DFM pastes are formulated with vegetable oil and inert gelling ingredients. Some fungal products are formulated with grain
by-products as carriers. Some DFM are designed for one-time dosing while other products are designed for feeding on a daily basis.
However, there is little information comparing the efficacy of administering a DFM in a single massive dose compared to continuous
daily dosing. Lee and Botts (1988) reported that pulse dosing alone or pulse dosing with daily feeding of Streptococcus faecium
M74 resulted in improved performance of incoming feedlot cattle. The need for a bacterial DFM to actually attach and colonize gut
surfaces in order to have a beneficial effect is also questionable. However, in certain applications, the argument could be made that
a DFM organism need only produce its active component (without colonization) to be beneficial. Dose levels of bacterial DFM have
varied. Studies can be found where L. acidophilus has been fed at levels ranging from 106 to 1010 cfu per animal per day.
Hutchenson et al. (1980) suggested that feeding more than 107 cfu per head per day may cause lower nutrient absorption due to
overpopulation of the gut. Orr et al. (1988) reported that feeding a continuous high dose of L. acidophilus to feeder calves (1010
cfu per head/day) had no effect on gain and actually reduced feed efficiency when compared to feeding a lower dose (106).
Tolerance of DFM microorganisms to heat is important since many feeds are pelleted. In general, most yeast, Lactobacillus,
Bifidobacterium, and Streptococcus are destroyed by heat during pelleting. In contrast, bacilli form stable endospores when
conditions for growth are unfavorable and are very resistant to heat, pH, moisture and disinfectants. Thus, bacilli are currently used
in many applications that require pelleting. Over-blending can sometimes compensate for microbial loss during pelleting, but this is
not an acceptable routine practice. Future improvements in strain development may allow use of heat sensitive organisms in pelleted
feeds. Bacterial products may or may not be compatible with use of traditional antibiotics and thus care should be taken when
formulations contain both types of additives. Information on DFM and antibiotic compatibility should be available from the
manufacturer. For example, some species of bacilli are sensitive to virginiamycin, and lactobacilli are sensitive to chlortetracycline
and penicillin. Viability of DFM products has improved over the past several years but it is highly advisable to adhere to storage
recommendations. For example, products should be kept away from moisture, excess heat, and light.
Table 3.1. Some bacteria that have potential uses as Probiotics

Sr. No. Organisms End Products or Potential Use

1. Lactobacillus acidophilus Lactic acid, acidophilin, glycosidases
2. Pediococsus acidilactici Pediocin (bacteriocin)
3. L. lactis Amylase, hydrogen peroxide, proteases
4. Bifidobacterium bifidum Ureases, lactic acid, formic acid
5. Bacillus subtilis Amylase, protease
6. Propionibacterium thoenii Propionicin PLG-1 (bacteriocin)
7. Megasphaera elsdenii Ruminal lactate utilizer
8. Propionibacteria sp. Ruminal lactate utilizer, propionate producer

Conclusion
Feeding of probiotics may have impact on the overall performance of the livestock and poultry. However, the results may not be
well pronounced always especially if the diets are adequate in all the nutrients and also the animals are not in stress condition.
Responses to a growth promoter depend on a variety of factors viz. product composition, malnutrition, stress condition, health of
animals, challenge from variant strains of pathogens etc. When these factors exist, probably the beneficial effects of probiotics could
be significant. Further, studies are needed to evaluate the probiotics with different combinations in different agroclimatic conditions.
In addition, detail studies should be carried out to develop standard packages comprising of ideal combinations of probiotics and
prebiotics (synbiotics) for obtaining optimum performance of livestock and poultry under different agro climatic conditions.

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 4 Prebiotics

Prebiotic foods have been consumed for centuries, either as natural components of food, or as fermented food. However, interest in
dietary use of prebiotics blossomed in the later 1800s and early 1900. Preliminary researches revealed that lactose had a profound
effect on the pH of the caecal contents and intestine due to lactic acid fermentation resulting in enhanced calcium and phosphorous
absorption. Further, it was also noted by several workers that inclusion of lactose in poultry diets resulted in better growth and
reduced mortality.
A prebiotic can be defined as “a non digestible food ingredient that beneficially affects the host by selectivity stimulating the growth
and/or activity of one or limited number of bacteria in the colon, and thus improves host health” (Gibson and Roberfroid, 1995).
Based on this definition, Russell (1998) formulated the criteria according to which a substance can be a prebiotic. First, prebiotics
are always feed ingredients that are not digested by the host, not or little used and/or metabolised as they pass through the upper
portion of the intestinal tract, so they can reach the flora of the large intestine. Secondly, they have to be able to serve as a substrate
for one or more bacterial species with a potentially beneficial effect on the host. Finally they have to be able to cause a shift in the
microflora that improves the health of the host. In principle only non-digestible, fermentable feed components are prebiotics.
Table 4.1. Intestinal functions assigned to prebiotics

Dietary fibers and gastrointestinal functions

Effects on upper GI • Resistance to digestion
tract
• Retarded gastric emptying
• Increased oro-caecal transit time
• Reduced glucose absorption and low glycaemic index
• Hyperplasia of the small intestinal epithelium
• Stimulation of secretion of intestinal hormonal peptides
Effects on lower GI • Acting as food for colonic microbiota
tract
• Acting as substrates for colonic fermentation
• Production of fermentation end products (mainly SCFAs)
• Stimulation of saccharolytic fermentation
• Acidification of the colonic content
• Hyperplasia of the colonic epithelium
• Stimulation of secretion of colonic hormonal peptides
 • Bulking effect on stool production
• Regularization of stool production (frequency and·
consistence)
• Acceleration of ceco-anal transit
A combination of probiotics and prebiotics is called a synbiotic (Collins and Gibson, 1999; Schrezenmeir and de Vrese, 2001). This
combination could improve the survival of the probiotic organism, because its specific substrate is available for fermentation.
Examples of synbiotics are FOS and bifido bacteria, and lactitol and lactobacilli.

Characteristics of Ideal Prebiotics

1. It should be neither hydrolysed nor absorbed by mammalian enzyme or tissues.
2. It must selectively enrich for one or a limited number of beneficial bacteria.
3. It should beneficially alter the intestinal microbiota and their activities.
4. It should beneficially alter luminal or systemic aspects of the host defense system.

Beneficial Effects of Prebiotics

There are several beneficial effects of prebiotics. They are as follows:
1. It increases biomass and stool bulking
2. It enhances vitamin-B synthesis.
3. It improves mineral absorption.
4. It increases production of VFA.
5. It prevents cancer
6. It lowers serum cholesterol
Besides these beneficial effects, prebiotics have several others, which have been illustrated in recent times. Prebiotics have been
found to minimize the incidence of ascites (Santos et al., 2005). Prebiotics have been also found to lower egg cholesterol (Chen et
al., 2005a), increase egg production and feed efficiency of layers (Chen et al., 2005b) and improve eggshell quality (Chen and Chen,
2004).

Classification
The term prebiotics is generally restricted to indigestible carbohydrates. These carbohydrates are divided in groups based on their
molecular length: mono di, oligo and polysaccharides.
The most important monosaccharide prebiotics are hexoses (glucose, fructose, galactose, mannose) and pentoses (ribose, xylose,
arabinose). Monosaccharides such as glucose and fructose are digestible as monomers and therefore not prebiotics according to the
definition of Russell (1998). Galactose is available mostly under the disaccharide feed additive. These monosaccharides can form the
basis for enzymatically constructed oligo or polysaccharides.
The most important natural disaccharides are sucrose, lactose and maltose. Isomerization products of these compounds can be used
as prebiotics e.g. lactulose (based on lactose). Lactose, lactulose and lactosucrose have prebiotic effects in chickens.
The most important polysaccharide prebiotic for chickens is guar gum, produced from the seeds of the guar bean, Cyamopsis
tetragonolobus. Another possible classification is based on their source of origin: natural or synthetic saccharides (Iji and Tivey,
1998). Examples of natural sources of oligosaccharides are soybeans, oil palm kernels, white lupin, blue lupin, chick pea and black
gram. The different classes of synthetic oligosaccharides are Lactulose, Mannan-oligosaccharides and Isomalto-oligosaccharides
etc. The list of different types of oligosaccharides is quite long. However only those that have been used in poultry have been
discussed in this paper.

Mechanism of Action of Prebiotics

Prebiotics can have a direct effect either by direct binding of pathogens or by increasing the osmotic value in the intestinal lumen.
More often, however, their effects are indirect, mediated by metabolites, which are generated by the intestinal flora that uses the
prebiotics for their own metabolism. Such metabolites include short-chain fatty acids, lactate, polyamines and bactericins.
Type 1 (F1) fimbria has been described in several bacterial enteric pathogens. These type I fimbriae bind to mannose residues of
glycoproteins, present on the surface of eukaryotic cells. They are needed for the adhesion of the bacteria to the mucosal surface,
which is a prerequisite for the colonization of the host. Indigestible carbohydrates with mannose residues may bind the F1 fimbriae
and therefore block the adhesion of the bacteria to the epithelial cells (Finucane et al., 1999). Prebiotics may constitute a substrate
for the growth of intestinal flora. This multiplication of normal flora may inhibit the colonization with pathogenic bacteria. This
phenomenon of inhibition is called “competitive exclusion”. Another possible mechanism of action of prebiotics is through a
modification of the metabolic activity of normal intestinal flora. Saccharides, indigestible by the host, are fermented by the flora into
volatile fatty acids (VFA including acetate, propionate and butyrate), lactate and several gases including carbondioxide, methane and
hydrogen. These acids lower the pH of the medium and may thus exert antibacterial effect. An added effect of acid production is
the protonation of potentially toxic ammonia (and amines) to produce NH4+, which is nondiffusable and thus lowers blood ammonia
levels.

Ruminants
The use of prebiotics in cattle has been limited due to the ability of ruminants to degrade most prebiotics; however enhancements in
rumen-protective technologies may allow these compounds to be used in feedlot and dairy cattle (Callaway et al., 2008), considering
also that several classes of nondigestible oligosaccharides are found in plant cell wall in nature including feeds normally used for
livestock feeding (Lema et al., 2002). Addition of MOS to the diet of Holstein calves improved fecal scores just as for antibiotic
treatment when compared to control milk replacer; whereas body weight was not affected (Heinrichs et al., 2003). Supplementation
of sorbitol, L-arabinose, trehalose, and rhamnose to cattle rumen medium displaced E. coli O157:H7 within 72 h (De Vaux et al.,
2002). The overall studies on the effect of forage and concentrate diets on fecal shedding and colonization of the gut by E. coli
O157:H17 are still unclear and little information is available; however the manipulation of the fiber content could bring new
perspectives maintaining the animals on a concentrate diet without sacrificing cattle weight gain as showed by Lema et al. (2002).

Pigs
TOS included at 35 g/kg in a diet for growing pigs resulted in a significant increase in fecal bifidobacteria and lactobacilli without
growth performance increase (Smiricky-Tjardes et al., 2003). A novel galactooligosaccharide (GOS) mixture, supplied at 40 g/kg
diet, resulted in a significant increase of the density of bifidobacteria and acetate concentration, and in a decrease of pH compared
with the control diet and a control diet supplemented with inulin. In addition, the oligosaccharide mixture strongly inhibited the
attachment of ETEC E. coli and S. enterica serotype typhimurium to HT29 cells in vitro (Tzortzis et al., 2005). An interesting study
was conducted on the effects of barley and oat cultivars, with different carbohydrate compositions, on the intestinal bacterial
communities in weaned piglets. Increased levels of â-glucans and altered amylopectin/amylose ratio seemed to selectively promote
butyrate-producing bacteria, able to hydrolyzed complex carbohydrates. Furthermore, bifidobacteria and lactobacilli counts were
positively affected by the choice of the cereal variety (Pieper et al., 2008). Oligosaccharides incorporated into swine diets at levels
ranging from 5 to 40 g/kg diet have resulted in mixed but generally not significant effects regarding beneficial modulation of microbial
populations determined in various intestinal segments and feces of swine (Mikkelsen et al., 2003). Mountzouris et al. (2006) showed
that the dietary treatment with fructooligosaccharides (FOS) or trans-galactooligosaccharides (TOS) did not influence the
populations of the beneficial bacterial but promoted saccharolytic activities in the porcine colon basing on the value of total volatile
fatty acids, acetate concentrations and glycolytic activities. Modesto et al. (2009) reported that GOS from milk whey, and sugar beet
fructooligosaccharides (sbFOS) added to the diet of weaned pigs in different amounts had no effect on the hindgut microbiota,
except for SbFOS at 40 g/kg which tended to increase the endogenous bifidobacteria, whereas growth performance was not
influenced.

Poultry
Lactose
Lactose is generally of animal origin and is found in the milk of mammals. Human milk contains 6% lactose and cows’ milk 4.5%.
Lactose is not hydrolyzed or absorbed intact from the intestinal tract of chicks, and as much as 50% of ingested lactose in poultry
diets may be excreted unchanged. Because of its lack of digestion and absorption, lactose passes into the lower segments of the
intestine and caeca. Lactose and its isomerization products such as lactulose reportedly have prebiotic effects in chickens.
Including lactose in the diet of chicken may produce significant improvement in body weight gain. It has been seen that lactose has a
hypocholesteromic effect in quails (Gujral, 2005). This may be due to enhanced activity of lactic acid bacteria requiring cholesterol
for their own metabolism resulting in decreased absorption of cholesterol in the gastro intestinal tract.
Mannan-oligosaccharide
Saccharomyces yeast outer cell wall components known as mannan-oligosaccharides (MOS) consists of Mannose and glucose
polymer. Commercially it is available as Bio-MosTM manufactured by Alltech Inc., USA. The product contains yeast cell wall
fragments derived from Saccharomyces cerevisiae. The cell wall fragments are obtained by centrifugation from a lysed yeast cell
culture. The pellet containing the yeast cell wall fragments is then washed and spray dried.
There are several reports available on studies with MOS in poultry. It has been suggested that MOS may be an alternative to
antibiotics in improving feed conversion (Fritts and Waldroup, 2000; Hulet et al., 2000). Supplementation of MOS may reduce
abdominal fat and increase breast yield in broilers. Further, MOS may reduce the ammonia concentration in the cecal digesta of
turkeys (Zdunczyk et al., 2005). It has been seen that MOS has immunomodulatory effect in birds. Inclusion of MOS @ 0.5g/ kg
significantly enhanced serum antibody titre against sheep red blood cells and increased macrophage activity in chickens (Cotter et
al., 2000; Shao et al., 2000). Savage and Zakrzewska (1997) reported increased concentration of bile IgA and plasma IgG in
response to mannanoligosaccharides in turkey poults. Similarly including MOS in the diets of broilers containing aflatoxins have led to
significant improvement in titre values against Newcastle disease and Infectious Bursal Disease (Swamy and Devegowda, 1998).
MOS has also been found to lower the egg total cholesterol, serum total cholesterol and LDL cholesterol in layers when provided
with 1.0 g/kg in the ration (Stanley and Sefton, 1999).

Fructo oligosaccharides
Fructo oligosaccharides (FOS) consist of a linear chain of B-D fructofuranose units linked 1, 2 by glycosidic bonds. Chains may
terminate with a D glycopyranose unit at the non-reducing end. They can be extracted from plants like onions and asparagus, by
controlled (limited) enzymatic hydrolysis of insulin polymers from chicory roots (Heinz and Vogel, 1991) and by enzymatic synthesis
from sucrose using a fructosyl transferase from Aspergillus niger or Aureobasidium pullalans (Hidaka et al., 1988; Hidaka and
Hirayama, 1991) according to the reaction:

Neosugar is a mixture of glucose, sucrose and FOS with a terminal glucose unit. Glucose and sucrose can be removed from the
reaction mixture by chromatography to obtain a product with increased FOS purity.
FOS has elicited a great interest to improve intestinal health and productivity in poultry. FOS is known for their ability to stimulate
growth of Bifidobacteria and to inhibit that of potentially pathogenic bacteria such as Enterobacteria, Clostridia and Salmonella.
FOS has shown to be resistant to intestinal glycolytic enzymes and to pass unaltered to the large intestine where they are fermented
by the microflora.
Significant improvement in weight gain, feed efficiency, mortality, carcass fat contents and dressing percentage have been observed
with addition of FOS in broiler diets. Further, feeding FOS to broilers may reduce intestinal colonization by pathogens in birds.
It has been seen that FOS enhances the growth of intestinal bacterial organisms especially Lactobacillus species and
Bifidobacterium. Some cellular components of Bifidobacteria promote immunological attack against malignant cells and thereby act
as immunomodulators. Further, FOS also aids in amelioration of antibiotic associated diarrhoea and reduction of serum triglycerides
and cholesterol. Growing female quails fed diet supplemented with FOS had higher lysozyme levels in blood serum (Szczerbinska et
al., 2000) and egg white with no reduction in serum protein content (Tarasewicz, 1998). Further, female quails supplemented with
FOS showed reduced contents of blood lipids and cholesterol (Tarasewicz, 1998). This may be due to the fact that acetate and
propionate in combination with L-lactate play a role in regulating lipid and cholesterol metabolism.

Miscellaneous prebiotics
Partially Hydrolysed Guargum (PHGG)
The most commonly used polysaccharide prebiotic for chickens is guargum, produced from the seeds of the guarbean, Cyamopsis
tetragonolobus. By selectivity cleaving the mannan back-bone chain of guar gum using endo-B-D-mannanase, a mixture of
galactomannans is obtained, called PHGG. Preliminary studies reveal that PHHG may reduce microbial load in birds.

Lactosucrose
Broiler chicks on a diet supplemented with lactosucrose at 1.5 g/kg have been shown to have a reduced incidence of lecithinase-
negative bacteria and lower concentrations of ammonia than those on the control diet (Terada et al., 1994). Further, lactosucrose
also increased the concentration of acetic and butyric acids in the ceca.

Isomalto oligosaccharide
Leuconostoc mesenteroides isomaltoolig saccharides (IMO) stimulated growth of Bifido bacterium and Lactobacillus and are not
used by Salmonella or E. coli cecal isolates. Further, Salmonella typhimurium grown in mixed cultures on IMO reduced the
Salmonella population. Cecal isolates grown on IMO showed higher viable counts and faster growth than Salmonella, indicating a
potential value for these oligomers for poultry intestinal micro flora modification (Chung and Day, 2004).

Conclusion
A wide variety of prebiotics is commercially available as feed additives. When added in limited amounts to poultry feeds, they can
result in a significant improvement in weight gain, efficiency of feed conversion and health status of the poultry. Further, it has been
seen that several herb polysaccharides may serve as alternative for antimicrobial growth promoters in chicken (Guo, 2003). Turkeys
are good forage feeders (Bhattacharyya et al., 2006). They are also resistant to different diseases unlike chicken in extensive or
semi-intensive management system. Hence, extracts of these polysaccharides may serve as a growth promoter in different species
of poultry. Prebiotics don’t have a major role to play when optimum conditions of management and housing are provided. However,
in practical conditions involving large flocks where there is deviation from the optimum conditions, prebiotics play a pivotal role in
cost involved and economic return.
Several key advantages of these prebiotics are as follows:
1. Most are natural products, made of very simple sugars, without any antigenic capacity.
2. They can promote directly the growth of beneficial bacteria already present in the intestine
3. They have no viability constraints like micro-organisms (probiotics).
4. They do not present side effects or accumulate in tissues like antibiotics.
5. They are resistant to high temperatures and to the acidic pH of the proventriculus, which avoids formulation and
application problems.
6. Their production cost is low and compatible with dose effect in poultry feed.
Elucidation of mechanism of action of each prebiotic will allow more accurate structure/function relationships to be established
leading to the design of specific carbohydrate structures with improved efficacy.

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 5 Enzymes

Feed accounts for a significant cost of production in livestock and poultry. In case of poultry, it represents 65% cost of the
production in broilers and 75% in layers. Presently the live stock industry faces a huge challenge due to the high cost of feed
ingredients and shortage of the conventional feed ingredients. Over the years, efforts have been made to use unconventional feed
ingredients. However, such feed ingredients have many anti nutritional factors that adversely affect the performance of livestock
and poultry. Due to the presence of these incriminating factors, the unconventional feed ingredients are poorly digested in livestock
and poultry. These poorly digested feed ingredients results in bacterial overgrowth, which becomes a potential substrate for bacterial
fermentation leading to intestinal disorders and disease. Application of antibiotics and other anti-microbials help to overcome the ill
effects. However, the ban on the use of sub therapeutic level of antibiotics in many countries has increased the relevance of use of
exogenous enzymes in the feed of livestock and poultry. These exogenous feed enzymes improve the nutritive value of different
feed stuffs, economize livestock and poultry rearing and reduce environmental pollution.
Enzymes are organic and biological catalysts that can initiate or accelerate biochemical reactions, converting one or more substrate
into products. In other words, enzymes are protein molecules that catalyze specific chemical reactions. Several exogenous enzymes
have been used in livestock and poultry to optimize the production. Besides the auto enzymatic digestion, ruminants have an added
advantage of alloenzymatic digestion provided by rumen microflora, which helps to obtain nutrients from complex feed unlike poultry
and other monogastric animals (Pariza and Cook, 2010). Hence, use of exogenous enzymes in poultry and other monogastric animals
holds significance from commercial point of view. Thus, several digestive enzymes have been studied for use as additives to
enhance animal performance with success in poultry and swine diets.

Poultry
Commercial exploitation of exogenous enzymes started a century back with patenting the process for production of alpha amylase
from the fungus Aspergillus oryzae (Pariza and Cook, 2010). Presently, most of the enzymes used in the food and beverage
industry are from Aspergillus except cellulase and hemicellulase derived from Trichoderma. Several enzymes have been exploited
or have a potential to be exploited in the poultry feed industry viz. phytases, proteases, lipases, cellulose (â-glucanases),
galactosidases, xylanases and associated enzymes (Table 5.1). However, the success of an enzyme depends on various factors viz.
the type of feed, anti nutritive factor in the feed, concentration and spectrum of an enzyme, type of bird, age of the birds etc.
Feed accounts for 65% of the cost of production in broilers and 75% in layers. Hence, optimum utilization of feed is very pivotal for
the best results in production economics. There are a lot of anti nutritive factors in poultry feed which minimizes the proper utilization
of feed. Supplementation of exogenous enzymes have been tried to counter act the adverse effect of the anti nutritive factors.

Phytates
Almost two thirds of the total phosphorous found in poultry feed of vegetable origin are found in the form of phytic acid. Phytic acid
is the hexaphosphate ester of myo-inositol. As birds lack phytase enzyme, phytates (calcium-magnesium-potassium salt) or phytic
acid is poorly utilized by birds and exceted by faeces. Hence, the available phosphorous or non phytin phosphorous is the difference
between total and unavailable or phytate phosphorous. It is calculated as plant P X0.30 + supplemental P + P from animal feed
origin. Thus, addition of exogenous phytase (Aspergiulls niger derived phytase feed enzyme) not only allows intestinal hydrolysis of
phytate and thereby increases the availability of phosphorous in feed but also reduces environmental pollution (Pariza and Cook,
2010).
Non starch polysaccharides (NSP)
The classification of non starch polysaccharides was initially based on the method used for extraction and isolation of
polysaccharides. The residue left after a series of alkaline extractions of cell wall materials was named cellulose and the fraction of
this residue that was solubilised by alkali was called hemicellulose. However, classification by differences in solubility lacks precision
with respect to both structure and function. Crude fibre (CF) refers to the residues of plant material after extraction with acid and
alkali and includes variable portions of the insoluble NSP. Neutral detergent fibre (NDF) refers to the insoluble portion of the NSP
plus lignin, and acid detergent fibre (ADF) refers to a portion of insoluble NSP comprised largely, but not exclusively, of cellulose
and lignin. Thus, the nutritional relevance of values obtained using these methods in monogastric nutrition are debatable. The
complexity in the structure, function and confusion in the nomenclature has made it almost impossible to draw a precise classification
of NSP. Practically, NSP may be classified into three main categories as shown below, namely cellulose, non-cellulosic polymers
and pectic polysaccharides (Bailey, 1973).
Non starch polysaccharides (NSP)

The NSP compounds are present in a variety of poultry feed stuffs viz. β glucans in barely and oat, arabinoxylans in wheat and
maize, arabinogalactans in rapeseed, galactomannans in guar, galactouronans and galactoarabinans in soybeans. These compounds
increase the viscosity, which reduce diffusion and contact with lipase and bile salt micelles. NSP decrease the digestibility of
carbohydrates, lipids and protein in poultry. NSP also increase the excreta volume and cause wet litter condition. Thus, a variety of
enzymes have been tried to counteract the toxic effects of NSP in poultry as shown in Table 5.1.
Table 5.1. Enzymes used in poultry feed

Enzyme Classification Production organism Function

Phytase Phosphatase Aspergillus sp., Penicillium sp., Hydrolyzes
Phytase canola, Pichia phytate
pastoris, d- Aspergillus niger,
d-Escherichia coli
Glucose oxidase Oxidoreductase Aspergillus niger Degrades glucose
to hydrogen
peroxide and
gluconic acid
Catalase Oxidoreductase Aspergillus niger var., Produces water
Micrococcus lysodeikticus and oxygen from
hydrogen
peroxide
Trypsin Protease Animal pancreas Hydrolyzes
proteins
Protease Protease Aspergillus niger, Aspergillus Hydrolyzes
sp., Bacillus sp. proteins
Pepsin Protease Animal stomachs Hydrolyzes
 proteins
Papain Protease Papaya (Carica papaya) Hydrolyzes
proteins
Keratinase Protease Bacillus licheniformis Hydrolyzes
proteins
Ficain Protease Ficus glabrata Hydrolyzes
proteins
Bromelain Protease Pineapple (Ananas cosmosus) Hydrolyzes
stem and fruit proteins
Lipase Lipase Aspergillus niger, Candida sp., Hydrolyze
Rhizomucor sp., Rhizopus sp. triglycerides,
diglycerides and
glycerol
monoesters
Xylanase Carbohydrase Aspergillus sp., Bacillus sp., Hydrolyzes
Humicola sp., Penicillum sp., xylans
Trichoderma
Pullulanase Carbohydrase Bacillus acidopullulyticus Hydrolyzes starch
Pectinase Carbohydrase Aspergillus aculeatus, Breaks down
Aspergillus niger, Rhizopus protein
oryzae
β-Mannanase Carbohydrase Aspergillus niger, Bacillus Hydrolyzes beta-
lentus Trichoderma reeseic mannans
Lactase Carbohydrase Aspergillus niger, Aspergillus Hydrolyzes
oryzae, Candida lactose to glucose
pseudotropicalis and galactose
Invertase Carbohydrase Aspergillus niger, Hydrolyzes
Saccharomyces sp. sucrose to
glucose and
fructose
Hemicellulase Carbohydrase Aspergillus sp., Bacillus sp., Breaks down
Humicola sp., Trichoderma sp hemicellulose
Glucoamylase Carbohydrase Aspergillus niger, Aspergillus Hydrolyzes starch
(amylosglucosidase) oryzae, Rhizopus niveus, with production
Rhizopus oryzae of glucose
β-Glucosidase Carbohydrase Aspergillus niger Hydrolyzes
cellulose
degradation
products to
 glucose
β-Glucanase Carbohydrase Aspergillus sp., Bacillus sp. Hydrolyzes β-
glucans
α-Galactosidase Carbohydrase Aspergillus niger, Morteirella Hydrolyzes
vinaceae var oligosaccharides
Saccharomyces
sp.
Cellulase Carbohydrase Aspergillus niger Breaks down
cellulose
α-Amylase Carbohydrase Barley malt Hydrolyzes starch
with production
of maltose
Maltogenic α- Carbohydrase Bacillus subtilis, d-Bacillus Hydrolyzes starch
amylase stearothermophilus with production
of maltose
α-Amylase Carbohydrase Aspergillus sp., Bacillus sp., Hydrolyzes starch
Rhizopus sp.
(Munir and Maqsood, 2013)

In the last century, use of enzymes in poultry feed took a new direction when it was reported that newly hatched chicks may be
deficient in digestive enzymes (Nitsan et al., 1991; Noy and Sklan, 1995 and Jin et al., 1998). Specific activities of lipase, amylase
and trypsin rapidly increase up to 2-3 weeks post hatch. Further, it has been suggested that the immaturity of the digestive system of
neonates may result in poor utilization of dietary nutrients (Jin et al., 1998). In addition, It has also been demonstrated that nutrient
digestion rather than the ability to absorb nutrients may be the primary limiting factor (Parsons, 2004). Therefore, dietary
supplementation of microbial lipase, amylase or protease enzymes not produced in sufficient quantities by chickens in juvenile stage
have been carried out with no significant improvement in body weight gain and FCR (Slominski et al., 2006).

Phytase
However, feeding enzyme preparations to improve ruminal digestion has been a questionable practice in the past. The reasoning
behind this thought came from the fact that enzymes are proteins and they would be subject to degradation by microbial proteases in
the rumen and/or inactivated by proteases in the small intestine.

Adding Enzymes (dry) to Animal Feed

Kopecny et al. (1987) reported that a cellulase enzyme complex was rapidly degraded by rumen bacterial proteases and addition to
ruminal fluid had no effect on in vitro fiber digestion. Some have suggested that feeding unprotected enzymes may be more useful
in immature ruminants where rumen microbial populations are not fully developed. For example, Baran and Kmet (1987) reported
that a pectinase-cellulase enzyme additive improved ruminal fermentation in newly weaned lambs but not in adult sheep (with
established rumen microflora). Recently, there has been renewed interest in the use of enzymes in ruminant diets because some
fibrolytic enzymes have been shown to be stable when incubated with protease enzymes. Fontes et al. (1995) reported that several
xylanases were resistant to several proteases but only one cellulase from a mesophilic organism was resistant to proteolytic attack.
Posttranslational glycosylation has also been reported to protect enzymes from deactivation caused by high temperatures and
proteinases (Olsen and Thomsen, 1991). Hirstov et al. (1998) reported that when added to the rumen, fibrolytic enzymes maintained
partial activity. However, integrity of the enzyme is not the only criteria that should be used when evaluating enzymes for ruminant
diets, because in order for them to be effective, they must bind to their substrate and catalyze reactions. Tricarico and Dawson
(1999) reported that the addition of xylanase and cellulase enzyme preparations improved the in vitro ruminal digestion of fescue
hay. Zinn and Salinas (1999) reported that a rumen-stable fibrolytic enzyme supplement increased the ruminal digestion of NDF and
Feed N by 23 and 5%, respectively. They also reported an improvement in dry matter intake and average daily gain in steers
supplemented with this additive. These data suggest that adding enzymes directly (in a dry form) to the diets of ruminants may
improve digestion and production.

Spraying Enzymes (liquid) Directly onto Animal Feeds

In the past use of enzymes was restricted to their application on to forages at the time of ensiling. However, this mode of application
has met with variable results. One method to protect or minimize enzyme degradation by ruminal proteases is to treat feeds with
enzymes just prior to feeding. When enzymes are applied to feed in this fashion, binding with substrates can cause conformational
changes that may help to protect these exogenous enzymes from ruminal degradation. Treacher and Hunt (1996) reviewed the use
of spraying enzymes directly onto feeds, rather than adding at the time of ensiling, to enhance their nutritive values. This approach
offers exciting possibilities for using enzymes to improve nutrient digestion, utilization, and productivity in ruminants and at the same
time reduce animal fecal material and pollution. Spraying enzymes onto feeds just before feeding provides increased management
flexibility for feeding and bypasses any negative interactions that the ensiling process may have on enzyme performance. A number
of different mechanisms have been theorized as reasons for positive effects including, direct hydrolysis, improvements in palatability,
changes in gut viscosity, complementary actions with ruminal enzymes, and changes in the site of digestion (Beauchemin and Rode,
1996; Treacher and Hunt, 1997). Feng et al. (1992) reported that pretreatment of dry grass with fibrolytic enzymes improved in vitro
ruminal fiber digestion. Lewis et al. (1996) reported that enzymes sprayed onto a grass hay: barley diet increased VFA production
and NDF digestion. Spraying enzymes on silage has increased the release of residual sugars and rate of NDF digestion. A growing
body of evidence exists that supports improvements in animal productivity when feeds are treated with enzymes prior to feeding. In
some instances, enzymes have been applied directly to the grain but in some studies enzymes were applied only to the forage
component of the diets prior to mixing into a TMR. Interestingly, several, but not all, publications have reported that high levels of
enzymes resulted in lower milk yields than moderate levels of enzyme treatment (Lewis et al., 1999; Beauchemin et al., 1995, Kung
et al., 2000). Over-treatment of feeds with enzymes may result in blocking sites that may otherwise be available for microbial
enzymatic digestion or may prevent attachment by rumen microbes. More research will be needed in this area.
Evaluating the activity of enzyme additives and predicting improvement in animal performance will be a challenge for future
research because temperature, time, substrate concentration, enzyme concentration, product reactions, cofactors, and pH, among
other factors, have profound effects on enzyme activity. In addition, sources (bacterial versus fungal) and activity of enzymes differs
markedly. The purity of enzyme products must also be ascertained because many commercial enzymes are actually complexes of
various enzymes that must work in concert to hydrolyze a substrate to monomer units. For example, crude preparations of a
cellulase enzyme complex actually contain endo- and exo-beta-1, 4 glucanases, beta-glucosidases, and cellobiase. Hemicellulase
preparations are even more complex. Determining the proper ratio of individual enzyme activities relative to the targeted feed must
be determined in order to optimize their effects on feeds. No universally accepted methods exist for determining enzyme activity but
they are usually based on release of a monomer under optimal and standardized conditions. Certainly, newer methods that evaluate
enzymes should consider their optimum activities at rumen temperature and pH.
We know very little about the stability of added enzymes and interactions of enzymes with components of feeds. If added during
processing, enzymes must be able to withstand temperatures during pelleting. Several practical problems must be addressed before
liquid enzymes will find acceptance on the farm. First, liquid enzymes will probably require refrigeration for prolonged storage and
thus bulk storage space will be needed. In addition, sprayer mechanisms must be mounted on to TMR wagons. The cost of
transporting liquid enzymes to farms will also be high because of the weight of the liquid.

References
Bailey, R.W. 1973. Structural Carbohydrates. In: Butler G.W. and Bailey R.W., eds., Chemistry and Biochemistry of Herbage. New
York: Academic Press, 1:157-211.
Baran, M. and Kmet, V. 1987. Effect of pectinase on rumen fermentation in sheep and lambs. Archives of Animal Nutrition. 7/8:
643.
Beauchemin, K. A., Rode, L.M. and Sewalt, V.J.H. 1995. Fibrolytic enzymes increase fiber digestibility and growth rate of steers
fed dry forages. Canadian Journal of Animal Science, 75: 641-644.
Beauchemin, K. A. and Rode, L.M. 1996. Use of feed enzymes in ruminant nutrition. Proceedings of the Canadian Society of
Animal Science Annual Meeting, Lethbridge, Alberta. 103-140.
Beauchemin, K. A., Yang, W.Z. and Rode, L.M. 1999. Effects of grain source and enzyme additive on site and extent of nutrient
digestion in dairy cows. Journal of Dairy Science, 82: 378-390.
Feng, P., Hunt, C.W., Julien, W.E., Dickinson, K. and Moen, T. 1992. Effect of enzyme addition on in situ and in vitro degradation
of mature cool-season grass forage. Journal of Animal Science, 70 (Suppl. 1): 309 (Abstract).
Fontes, C.M. G. A., Hall, J., Hirst, B.H., Hazelwood, G.P. and Gilbert, H. J. 1995. The resistance of cellulases and xylanases to
 proteolytic inactivation. Applied Microbiology and Biotechnology, 43: 52-57.
Fuller, R. 1989. Probiotics in man and animals. Journal of Applied Bacteriology, 66: 365-378.
Hirstov, A., McAllister, T. A. and Cheng, K. J. 1998. Effect of dietary or abomasal supplementation of exogenous polysaccharide-
degrading enzymes on rumen fermentation and nutrient digestibility. Journal of Animal Science, 76: 3146-3156.
Jin, S. H., Corless, A. and Sell, J. L. 1998. Digestive system development in post-hatch poultry. World’s Poultry Science Journal ,
54: 335-345.
Kopency, J., Marounek, M. and Holub, K. 1987. Testing the suitability of the addition of Trichoderma viride cellulases to feed
rations for ruminants. Zivocisna Vyroba, 32: 587.
Kung, L., Jr., Treacher, R. J., Nauman, G. A., Smagala, A.M., Endres, K.M. and Cohen, M. A. 2000. The effect of treating forages
with fibrolytic enzymes on its nutritive value and lactation performance of dairy cows. Journal of Dairy Science, 83: 115-
122.
Lewis, G. E., Hunt, C.W., Sanchez, W. K., Treacher, R., Pritchard, G. T. and Feng, P. 1996. Effect of direct-fed fibrolytic enzymes
on the digestive characteristics of a forage-based diet fed to beef steers. Journal of Animal Science, 74: 3020-3028.
Lewis, G. E., Sanchez, W. K., Hunt, C.W., Guy, M. A., Prichard, G.T., Swanson, B. I. and Treacher, R.J. 1999. Effect of direct-fed
fibrolytic enzymes on the lactational performance of dairy cows. Journal of Dairy Science, 82: 611-617.
Munir, K. and Maqsood, S. 2013. A review on role of exogenous enzyme supplementation in poultry production. Emirates Journal
of Food and Agriculture, 25: 66-80.
Nitsan, Z., Duntington, E.A. and Siegel, P.B. 1991. Organ growth and digestive enzyme levels to fifteen days of age in lines of
chickens differing in body weight. Poultry Science, 70: 2040-2048.
Noy, Y. and Sklan. D. 1995. Digestion and absorption in the chick. Poultry Science, 74:366-373.
Olsen, O. and Thomsen, K. K. 1991. Improvement of bacterial b-glucanase thermostability by glycosylation. Journal of General
Microbiology, 137: 579-585.
Orr, C. L., Ware, D. R., Manfredi, E. T. and Hutchenson, D. P. 1988. The effect of continuous feeding of Lactobacillus
acidophilus strain BT1386 on gain and feed efficiency of feeder calves. Journal of Animal Science, 66 (Suppl. 1): 460
(Abstract).
Pariza, M.W. and Cook, M. 2010. Determining the safety of enzymes used in animal feed. Regulatory Toxicology and
Pharmacology, 56: 332-342.
Parsons, C.M. 2004. Gastrointestinal development and nutrient digestion in chicks, in: Proceedings of the 25th Western Nutrition
Conference, Saskatoon, Canada, 169-176.
Rode, L. M., Yanf, W. Z. and Beauchemin, K. A. 1999. Fibrolytic enzyme supplements for dairy cows in early lactation. Journal of
Dairy Science, 82: 2121-2126.
Schingoethe, D. J., Stegeman, G. A. and Treacher, R. J. 1999. Response of lactating dairy cows to a cellulase and xylanase enzyme
mixture applied to forages at the time of feeding. Journal of Dairy Science, 82: 996-1003.
Slominski, B.A., Meng, X., Jia, W., Guenter, W. and Jones, O. 2006. The effect of lipase, amylase and protease addition on growth
performance and nutrient digestion in young broiler chickens fed corn-soybean meal diet. Proceedings of the 12th European
Poultry Conference, Sept. 10-14, Verona, Italy (CD ROM).
Treacher, R. J. and Hunt, C.W. 1996. Recent developments in feed enzymes for ruminants. Proceedings of Pacific Northwest
Nutrition Conference. Seattle, WA.
Tricarico, J. M. and Dawson, K. A. 1999. Effects of defined xylanase and cellulase enzyme preparations on digestive processes of
ruminal microbial cultures. Journal of Dairy Science, 77 (Suppl. 1): 252. (Abstract)
Yang, W. Z., Beauchemin, K. A. and Rode, L. M. 1999. Effects of an enzyme feed additive on extent of digestion and milk
production of lactating dairy cows. Journal of Dairy Science, 82: 391 403.
Zinn, R. A. and Salinas, J. 1999. Influence of Fibrozyme on digestive function and growth performance of feedlot steers fed a 78%
concentrate growing diet. Proceedings of Alltech 15th Annual Symposium. Biotechnology in the Feed Industry. Nottingham
University Press. 313-319.
 6 Organic Acids

An organic acid is an organic compound with acidic properties. The most common organic acids are the carboxylic acids, whose
acidity is associated with their carboxyl group –COOH. However, sulphuric acids, containing the group –SO 2OH, are relatively
stronger acids. Alcohols, with –OH, may also act as weak acids. Some other groups may also act as weak acids viz. thiol group and
phenol group. In biological systems, organic compounds containing these groups are generally classified under organic acids. Some
common organic acids are lactic acid, acetic acid, formic acid, citric acid, oxalic acid, uric acid, propionic acid and butyric acid.
Organic acids have been used for decades in commercial compound feeds, mostly for feed preservation, for which formic and
propionic acids are particularly effective. Since the ban of antibiotic growth promoters in many countries, organic acids have been
used increasingly, not only because of their preservative qualities, but also for their nutritive properties. Organic acids and their salts
are generally regarded as safe and have been approved by most member states of EU to be used as the feed additives in animal
production.
Organic acids are both bacteriostatic and bactericidal. As un-dissociated organic acids are lipophilic, they can cross the cell
membrane of Gram negative bacteria, such as Salmonella. Once inside the cell, the higher cytosolic pH causes the acid to
dissociate, releasing hydrogen ions, which consequently reduces the intracellular pH. Microbial metabolism is dependent on enzyme
activity, which is depressed at lower pH. To redress the balance, the cell is forced to use energy to expel protons out across the
membrane via the H+-ATPase pump to restore the cytoplasmic pH to normal. Over a period of exposure to an organic acid, this can
be sufficient to kill cell. Expelling protons also leads to an accumulation of acid anions in the cell (Lambert and Stratford, 1998),
which inhibits intracellular metabolic reactions, including the synthesis of macromolecules, and disrupts internal membranes. Lactic
acid bacteria are less sensitive to the pH differential across the cell membrane, and thus remain unaffected. Inhibition of microbial
growth by this mode of action has been exploited for thousands of years in food preservation; organic acids are natural by-products
of microbial metabolism.

Poultry
Organic acids in un-dissociated (non ionised, more lipophilic) form can penetrate the bacteria cell wall and disrupt the normal
physiology of certain types of bacteria. Besides antimicrobial activity, they reduce the pH of digesta and increase the pancreatic
secretion. Acidification with various organic acids reduces the production of toxic components by the bacteria and colonization of
pathogens on the intestinal wall, thus preventing the damage to epithelial cells. Organic acids improve the digestibility of proteins,
calcium, phosphorus, magnesium, and zinc. Thus, they enhance nutrient utilization leading to better growth and feed conversion
efficiency.
Organic acid are lipid soluble in the un-dissociated form in which they are able to enter the microbial call. However carrier mediated
transport mechanism seem to be also involved in the membrane transportation.

Butyric acid
Butyric acid may improve growth, feed efficiency and GIT health. Butyric acid given to broiler chicks may influence body weight
gain and feed conversion ratio. It has been observed that pH of gastro intestinal tract (GIT) is reduced by butyric acid
supplementation (Leeson et al., 2005).
Butyric acid may improve dressing percentage and reduce abdominal fat in commercial broilers. Further, butyric acid may play a
role in the development of intestinal epithelium (Rama Rao et al., 2003).
Acetic acid
Acetic acid improves the heath of chicken due to higher production of pancreatic enzyme, which improves GIT health. Dietary
inclusion of acetic acid may improve FCR, bodyweight and carcass yield in broilers (Denli et al., 2003; Abdel Fattah et al., 2008).

Citric acid
Citric acid reduces PH of digesta, may improve gut health and degrades aflatoxin in the poultry ration. In addition, citric acid may
also improve poultry performance by reducing colonization of pathogenic micro-organism and toxic bacterial metabolites such as
ammonia and amines. Citric acid improves F.C.R, gut health immunity and live body weight in broilers (Chaveerach et al., 2004;
Mendez Albores et al., 2005).

Propionic acid
Propionic acid may affect the integrity of microbial cell membrane or cell macro molecules or interfere with nutrient transport and
energy metabolism causing the bactericidal affect. Propionic acid has PH reducing property. It also improves gut health, FCR,
immunity in broiler chicks at young stage (Dibner and Buttin, 2002).

Lactic acid
Lactic acid bacteria break down lactose (milk sugar) into glucose and galactose. The acidic environment inhibits the growth of
harmful bacteria. Lactic acid bacteria plays a role in absorption of vitamins D and K and formation of soluble salts of calcium and
iron in broilers (Schingoethe, 1976 and Morishita et al., 1982).

Role of organic acid in egg production and quality parameters

Phosphorus (P) is an essential mineral for metabolism and egg production of layers. It has been observed that organic acid
supplementation at 780 ppm maintained production end till 70 weeks of age in layers. Organic acid diet increases egg shell quality,
egg shell protein, egg shell calcium, improve mineral and protein absorption. It has also been reported that organic acid in diet
increases hen day egg production and hen house egg production in layers (Zeidler, 2001; Rodriguez Navarro et al., 2002 and Chen
and Chen, 2004).

Role of organic acid in immunity of poultry

The first 3 weeks of life comprise a critical period in chick’s life for gut maturation. During this period healthy birds perform at
maximum potential without diverting energy towards immune response processes (Klasing, 2007; Yegani and Korver, 2008 and Yin
et al., 2010). Gut morphology affects nutrient absorption, and greater villi length and crypt depth are associated with functional ability
(Stokes et al., 2001; Yang et al., 2007 and Choct, 2009). Commensal bacteria of the gut benefit the host in a variety of ways,
including organic acid production and immunomodulation (Tse and Chadee, 1991). Organic acids inhibit pathogenic bacteria growth
by disrupting bacteria cell membrane transport, preventing the bacteria from reaching equilibrium with their environment
(Cherrington et al., 1990). Probiotics, beneficial microbial cultures, may be administered to stimulate the local immune system
(Fuller, 1989; Gibson and Roberfroid, 1995; Netherwood et al., 1999) and enhance epithelial innate immunity-related gene expression
through anti-inflammatory effects and reduced pro-inflammatory cytokine expression such as IL-6 (Pagnini et al., 2010).Therefore,
the organic acids pay a pivotal role on intestinal morphology and innate immunological responses of chickens during this period. It
has been reported that level of gamma globulin increased in broilers with dietary inclusion of organic acids (Rahmani and Speer,
2005).

Pigs
Addition of organic acids to feed combats susceptible microorganisms, including pathogenic bacteria and some fungi, which would
otherwise cause spoilage and reduce its nutritive value by metabolizing the starch and protein therein. In pig diets, organic acids and
their salts also take effect in the gastrointestinal tract, mainly in the proximal tract - the stomach and small intestine. Firstly, organic
acids lower the pH of the stomach contents, which can especially beneficial at weaning, where it stimulates the conversion of
inactive pepsinogen to active pepsin. This may improve protein digestibility and decrease the rate of gastric emptying. Organic acids
also stimulate exocrine pancreatic secretion of enzymes and bicarbonate, thus assisting with protein and fat digestion. Furthermore,
organic acid anions can complex with calcium, phosphorus, magnesium and zinc, improving the digestion of these minerals and
reducing the excretion of supplemental minerals and nitrogen (Roth et al., 1998a, b). The bacteriostatic or bactericidal effects of
organic acid anions also take effect in the proximal gastrointestinal tract. It should be noted that whereas organic acids lower gastric
pH, organic acid salts do not (Eidelsburger et al., 1992a). Therefore, the improvements in growth performance resulting from dietary
inclusion of organic acid salts are due to an antimicrobial effect. Butyric acid for instance, is the main energy source for the epithelial
cells of the large intestine and is considered to be effective for promoting epithelial growth (Galfi and Bokori, 1990).

Organic acid on pig performance

At weaning, piglets are particularly susceptible to infection with intestinal pathogens, as well as being inadequately equipped
physiologically to deal with solid feed. The buffering capacity of weaning feeds is also high, compounding the problem through a
negative effect on pepsin activity in the stomach (Eidelsburger et al., 1992b), a problem that is addressed through the acidification of
diet. In the diet of grower finisher pigs, the anti-microbial effect of organic acids in the feed (hygiene), stomach and small intestine is
largely responsible for their performance enhancing benefits. Effective doses have been established that can improve productivity of
pigs to levels comparable with antibiotic growth promoters (Øverland et al., 2000).
Most recently it has been studied adding organic acids to diets for sows. Øverland et al. (2009) added 0.8% or 1.2% potassium
diformate to diets for primiparous and multiparous sows from one mating to ting of acidifiers the next, feeding of acidifier through
gestation and lactation. The performance of the piglets of these sows was also recorded and compared. The authors found that
sows fed potassium diformate had increased back fat thickness during gestation, although daily feed intake and body weight gain did
not change. Feeding potassium diformate also tended to be associated with a heavier birth weight of piglets, irrespective of dose. It
also improved average daily gain, resulting in a greater weaning weight. Sows fed the diets containing potassium diformate tended to
have increased milk fat content on day 12 post-farrowing. On the other hand, sows fed potassium diformate at a dosage of 0.8%
under tropical conditions (Lückstädt, 2011) tended (P < 0.1) to have a higher feed intake from 3 days after farrowing onwards.
Furthermore, reduced weight loss (P = 0.06) during the weaning period and lower back fat loss (P = 0.05) was observed.

Salmonella control in fattening pigs

Good gut health is increasingly being shown to be effective against intestinal pathogens, a strategy that has only been made possible
through the removal of antibiotic growth promoters in feed. Creating and maintaining a healthy intestinal environment has become
essential to productivity and food safety programmes alike. Salmonella enteritica Typhimurium is the predominant serotype found
in pig carcasses, accounting for about 71% of cases. Several serotypes are resistant to antibiotics, which has put pressure on
producers to prevent contamination. While salmonella cannot be wholly eradicated in pig units, it can be controlled to minimise the
risk to consumers. Biosecurity plays a significant role in salmonella control. In feed compounding, although heat treatment is
effective in reducing contamination of feed leaving the feed mill, this effect does not persist during transport, storage and subsequent
outfeeding. When conditions within the feed are less conducive to bacterial infection, salmonella contamination can be reduced. The
next critical control point is within the pig’s gut, where conditions for bacterial growth may again be optimal. Salmonella growth
requires warmth (35- 370 C is optimal), a moisture content greater than 12% and a pH between 4.5 and 9.0. It is no coincidence that
the pig gut can provide salmonella everything they need to thrive.
A study by Dennis and Blanchard (2004) in the UK and a more recent study by Correge et al. (2010) in France also concluded that
potassium diformate is an effective tool in salmonella control strategies on commercial farms, as it reduced the percentage of
salmonella-positive pigs by 50% and decreased salmonella ELISA scores in pork meat juice by 46% in grower finisher pigs. The UK
trial also showed an improvement in daily gain of 7.7%, reduced mortality and a reduction in medicinal intervention compared with
the rolling average for the unit, which showed an economic benefit for implementing salmonella control measures.

Ruminants
Very few researches have been conducted to evaluate the effects of organic acids on ruminant performance. Kung et al. (1982)
reported that feeding 140 g of malate per day increased milk persistency in lactating cows and increased total VFA during early
lactation. Feeding malate to Holstein bull calves improved ADG and feed efficiency but had little effect on blood serum constituents
(Sanson and Stallcup, 1984). Even though in vitro studies have shown that DL-malate favorably alters ruminal fermentation (Martin
and Streeter, 1995; Callaway and Martin, 1996), little information is available detailing the effects of DL-malate on beef cattle
performance. In another studies, the addition of malic acid into ruminant concentrates is reported to have a positive effect on daily
live weight gain (Martin et al. 1999; Mungói, 2007a) whereas in others (Carro et al. 2006; Mungói, 2007b), no change in the daily
weight gain was found. Carro et al. (2006) stated that the addition of malic acid into lamb rations didn’t affect forage and
concentrate intake, but Mungói (2007b) stated that the forage intake increased and the concentrate intake decreased according to
the gradual incorporations of malic acid into concentrates for lambs during the growth period. With the addition of malic acid into
ruminant food, the changes in the rumen fermentation products and ruminal pHs are reported to be similar to the effects of
ionophores (Martin and Streeter, 1995). Martin and Streeter (1995) stated that the total volatile fatty acids and the ammonia nitrogen
contents in the rumen from cows supplemented with DL malate yielded similar results with not supplemented controls. However,
Mungói (2007b) reported that the total volatile fatty acids increased in ruminants supplemented with organic acids.

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 7 Fatty Acids

Fats are rich sources of energy. Fats provide 2.25 times more available enrgy than carbohydrates. Fats also increase the palatability
and reduce the dustiness of feed. Fats and fatty acids decrease the heat increment in the body. Fatty acids are also involved in
number of physiological functions of the body. Recently, considerable interest has been shown on studies pertaining to dietary
supplementation of polyunsaturated fatty acids (PUFA) in animal feed and their role in animal health and production.

Poultry
Nutrition can modulate quantitative and qualitative aspects of the immune response to pathogens. Research on poultry, especially in
chickens has elucidated the impact of diet on immunocompetence (Cook, 1991; Koutsos and Klasing, 2001) and the mechanisms that
are responsible. Elevated levels of fat-soluble vitamins and omega-6 fatty acids may influence the hatchling’s early inflammatory and
immune response. The fatty acid composition of diet affects the fatty acid composition of the yolk, which in turn can affect
embryonic development and hatchability. Palmitic acid increase yolk oleic acid levels to affect transport across the yolk sac
membrane and spare linoleic acid for embryonic growth and hatchability. Adding corn oil, palmitic acid, oleic acid or linoleic acid to a
basal diet increased hatchability and decreased late embryonic mortality (Vilchez et al., 1992). It has also been observed that
maternal dietary lipids alter bone development by influencing organic matrix quality and mineralisation in embryos (Liu et al., 2003).
A redirection of nutrient flow to meet the metabolic requirements of an immune response or inflammatory reaction is referred to as
homeorhesis (Bauman and Currie, 1980). This is in clear contrast to the homeostatic mechanism of maintaining metabolic
equilibrium. In fact it is the need of the hour to study if dietary manipulation can promote disease resistance and immunity. Protective
immune responses require a supply of nutrients at the appropriate times and amounts (Humphrey et al., 2002). While amino acids
are needed as substrates to aid the production of immunoglobulins, lysozymes, complement, cytokines, monocytes, heterophils and
clonal proliferation of antigen driven lymphocytes; fatty acids bind to intracellular receptors or modify the release of secondary
messengers. PUFA modulates the intercellular communication by regulating the incorporation of fatty acids into cell membranes
(Korver and Klasing, 1995, 1997). Linoleic acid is elongated to arachidonic acid and incorporated into cell membranes. Arachdonate
on release is converted into prostaglandins, leukotrienes and thromboxanes. Similarly, the n-3 PUFA can be incorporated into cell
membrane and on release determines the amount and type of eicosanoids and thus regulates cell communication. Increases in
antibody responses to antigens and decreases in mitogen-induced proliferation of lymphocytes are due to modulatory effects of
dietary n-3 fatty acids (Fritsche et al., 1991; Korner and Klasing, 1997).
Fatty acids have a profound role on the development of immune system. Hamdy et al. (2003) reported that supplementation of
sunflower oil along with linseed oil in broiler chicks significantly increased differential leukocyte counts and the relative weight of the
bursa of fabricius but there were changes in the relative weights of spleen and thymus.
Cheng et al. (2003) observed that anti-bovine serum albumin (BSA) antibody titres in laying hens fed fish oil or linseed oil were
higher compared to hens fed the control diet. Selvaraj and Cherian (2004) suggested that n-3 fatty acids increased production
performances and antibody mediated responses, while n-6 fatty acids and conjugated linoleic acid increased cell mediated responses
in broiler birds. However, Friedman and Sklan (1997) found that in turkeys vaccinated against Newcastle, infectious bronchitis and
necrotic enteritis, specific antibody response was related quadratically to serum linoleic acid and total n-6 polyunsaturated fatty acid
concentration. No correlation was found with linoleic acid or arachdonic acids. Similarly, Fritsche and Cassity (1992) concluded that
n-3 dietary fatty acids did not alter the primary or secondary antibody response of broiler chickens to sheep red blood cells. Rather
n-3 fatty acids reduced antibody dependent cell cytotoxicity and altered eicosanoid release by chicken immune cells.
In fact, the ratio of n-3 to n-6 PUFA plays an important role in modulating humoral and cell mediated immune response. Wang et al.
(2000) observed that a linseed oil diet increased the IgG concentration in laying hen serum. Sunflower oil reduced IgY content in egg
yolk. Torki et al. (2003) reported that antibody production against sheep red blood cells in chicks fed a diet with a higher ratio of n-3:
n-6 PUFA was higher among the dietary groups where as moderate dietary ratio of n-3: n-6 PUFA improved antibody production
against Newcastle disease virus. Similarly, Xia et al. (2003) found that antibody titers in fish oil and linseed oil supplemented diets
were higher than that in laying hens fed corn oil. The proliferation response to concanavalin was lower in laying hens that were fed
oils rich in n-3 fatty acids. They also suggested that higher level of n-3 fatty acids could improve immune functions of laying hens.
Thus, they concluded that dietary fat source and level had a significant impact on immune responses of laying hens.
Modulation of the magnitude and isotype of antibody responses of poultry to T cell-dependent antigens is affected not only by type of
essential fatty acids but also by their source. Higher total antibody and IgG titres to bovine serum albumin were found especially
after primary immunization in pullets given the sunflower oil enriched diet. Birds given sunflower oil mounted significantly lower IgM
titres to bovine serum albumin after primary and secondary immunization (Parmentier et al., 2002).
Sijben (2002) suggested that inclusion of 1-2% of n-3 and inclusion of no more than 3-4% of n-6 in the diet is optimal for antibody
responsiveness, enhances T cell reactivity, and possibly improves chicken’s disease resistance. Zaki and Hady (1995) concluded that
different dietary fat sources have different impact on performance and immune response of broiler chickens. While a mixture of
beef tallow and linseed oil (1:1) when fed to male Cobb broiler chicks significantly increased body weight compared with those fed
on the fat sources separately, haemagglutinin antibody titre and delayed hypersensitivity reaction (to phytohaemagglutinin-P) values
were higher in chickens fed on linseed oil. Though thymus and spleen weights were not affected by the inclusion of different fat
sources, bursal weight was significantly higher at 3 and 7 weeks of age in chickens fed on linseed oil.

Ruminants
Milk yields of high-producing dairy cattle demand that energy intake be maximized. Energy intake is not only important during the
period of peak milk production but also must be sufficient to maintain persistency of production and support body weight gain
following peak milk yield. To increase ration energy density, cereal grains are used to replace forages, but this practice is limited
since a certain amount of effective fiber is required to optimize ruminal fermentation, enhance nutrient digestibility, and maintain DM
intake. Recent research indicates that low forage diets can be fed if starch digestion is monitored by considering source of starch,
processing and particle size of the cereal grain, and nonforage fiber sources are used to dilute starch from the ration (Firkins et al.,
2001). Because fat is much higher in energy per unit of weight than cereal grains, it is used to increase energy density of diets.
Digestion and absorption of fat, suggested feeding guidelines for various fat sources, and the effects of feeding supplemental fat on
milk composition will be discussed in this chapter.

Biohydrogenation and Digestion of Fat

Fat digestion begins in the rumen (Figure 7.1). Bacterial lipases, and possibly to a minor extent plant lipases, break down fat into
glycerol and fatty acids (FA). Glycerol is used as an energy source by bacteria and is principally converted to propionic acid. The
unsaturated FA are biohydrogenated by bacteria, but the bacteria do not use them as an energy source. Most of the FA flowing to
the small intestine, where absorption takes place, will be free FA but some diglycerides, monoglycerides, and microbial phospholipids
will reach the small intestine. Presence of pacreatic lipase will result in cleavage of the glycerides and phospholipids so the FA can
be incorporated into the micelles for absorption. With the nonpolar nature of lipids, bile is very critical for emulsification of the FA
(for their incorporation into micelles) so that absorption can take place. The FA are reassembled as phospholipids and glycerides and
packaged as very low density lipoproteins (VLDL) and chylomicrons that have an outer protein matrix to aid in the transport of lipids
throughout the lympathic system.
 Fig 7.1: Digestion and absorption of fat in ruminants
The desirable characteristics of a fat source are that it should have minimum effects on ruminal fermentation and have a high
digestibility; however, these two qualities are not always easy to achieve. Unsaturated fat is highly digestible but may reduce fiber
digestibility in the rumen due to inhibitory effects on cellulolytic microorganisms. Saturated fat has less influence on fiber digestion in
the rumen, but digestibility of the fat may be inferior depending on the level of saturation (Firkins and Eastridge, 1994; NRC, 2001).
When comparing fats based on saturation level, iodine value (IV) or total unsaturates can be used. Iodine value is more reflective of
level of unsaturation (or saturation) because the relative proportion of unsaturated fatty acids (e.g. C18:1 versus C18:2 versus C18:3)
is not reflected within total unsaturates.
The potential negative impact of unsaturated fat from oilseeds may be minimized if the oilseeds are fed either whole or coarsely
cracked rather than extruded (Faldet and Satter, 1991; Reddy et al., 1994). This would allow the encapsulated oil to be released at a
slower rate in the rumen, or some of the oil may even escape the rumen. Because of fat being nonpolar (hydrophobic), it seeks out
particulate matter in the aqueous environment in the rumen, especially attaches to the waxy layer of forages. Free oil from oilseeds
should not be added to ruminant diets because of the rapid contact of the oil with particulate matter and microbes. Grinding soybeans
should not be done because of reduced rumen undegraded protein in roasted soybeans (Tice et al., 1993) and the potential increase
in oxidative rancidity with ground raw soybeans (unless grinding is frequent).

Fat Sources
Fat sources can be grouped into two major categories: natural fats and commercial fats. The natural fats can be sub-divided into
plant and animal fats. Commercial fats are special preparations made by using animal or plant fats. The chemical and fatty acid
compositions of various fat sources are provided in Table 1.

Natural fats
Oilseeds are the major source of fat from plants. Whole cottonseed is a well-balanced feedstuff for dairy cattle because it is
relatively high in protein, fiber, and energy. Due to physical characteristics of cottonseed, especially linted cottonseed, they are most
easily handled by inclusion in a total-mixed ration. However, the availability of Easiflo (coated with 2.5% starch to mat the linters)
and pelleted cottonseed provide for increased potential usage. Whole cottonseed contains the pigment gossypol that is toxic to
animals, especially non-ruminants. With the levels of cottonseed typically consumed by dairy cattle (< 15% of DM), intake of
sufficient gossypol to cause health problems is not likely (Coppock et al., 1987). However, cottonseed should be closely monitored
for mycotoxin contamination, especially aflatoxin.
Raw soybeans should not be ground and added to feed mixtures containing urea because soybeans contain urease. Although the
trypsin inhibitor in soybeans is a concern when feeding them to non-ruminants, it is assumed that most of the trypsin inhibitor is
deactivated in the rumen. On the other hand, some of the trypsin inhibitor may escape ruminal fermentation and decrease crude
protein digestibility in the intestines (Palmquist and Conrad, 1971; Tice et al., 1993). Soybeans can be included in total-mixed rations,
top dressed, or in the case of cracked beans (whole seeds will separate out), can be added to grain mixtures. As alluded to earlier,
exposure of oil and release of lipoxygenase by grinding raw soybeans may lead to rancidity problems, especially during summer
months. Potential for the oil to become rancid is greater when the ground soybeans are added to ensiled forages or wet by-products
during warm months.
Roasting soybeans will denature the urease, trypsin inhibitor, and lipoxygenase and decrease ruminal degradability of the protein. For
example, protein degradability in raw soybeans is about 72% whereas protein degradability in roasted soybeans is about 50%.
Quality of the roasting can be quite variable from one roaster to another with such factors as temperature, moisture level of seed,
rate of transit through roaster, and handling of beans after exiting the roaster giving rise to some of the quality differences. A few
beans should be broken and examined for uniformness of heat penetration - beans should not be raw in the center. Over-heating of
beans will reduce protein digestibility; therefore, occurrence of charred seeds should be avoided. Although visual appraisal alone is
not adequate for assessing quality of the roasting, seeds should be light brown in color. The protein dispersibility index (Hsu and
Satter, 1995) is available from some labs as a measure of the adequacy of heat processing of soybeans.
Other oilseeds such as canola, safflower seeds, and sunflower seeds can also be fed to dairy cattle. Caution should be exercised in
feeding these oilseeds because the oil is highly unsaturated, and they are higher in oil than cottonseed and soybeans, resulting in
lower recommended feeding levels. For example, only about one-half as much canola (seeds should be cracked) can be fed as
cottonseed or soybeans.
The primary animal fat fed to dairy cattle is tallow. Tallow contains more saturated fatty acids than the oilseeds, but handling is more
difficult because it is solid or semi-solid at room temperature. Tallow can be readily purchased in barrels with heating instruments to
melt the fat for mixing purposes. Tallow can be of different qualities, and some of the grades are as follows: edible tallow, extra
fancy tallow, fancy tallow, bleachable fancy tallow, and prime tallow. The different grades refer to the purity/cleanliness of the
tallow, and some of the grades may contain appreciable amounts of lard.
Yellow grease is waste grease from food service operations, and it may contain variable mixtures of vegetable and animal fats.
Yellow grease is used as a fat for livestock and pet foods, as an industrial raw material, and as a diluent in higher grade inedible fat
products such as bleachable fancy tallow. Several animal-vegetable fat blends also are available for feeding to dairy cattle.
Several other feedstuffs (e.g. hominy, dry distillers grain with solubles, and fish meal) contain a moderate amount of fat. Total fat
from all sources in diets should be the focus instead of only supplemental fat from major contributors. Because fish meal contains an
appreciable amount of 20 and 22-carbon polyunsaturated FA which are very toxic to ruminal bacteria (Hoover et al., 1989), it should
be restricted to a maximum 2 to 3% of dietary DM.

Commercial fats
Several commercial fat preparations are available and most of them are marketed as rumen inert sources. These fats fall into two
general categories: calcium salts and processed tallow (either hydrolyzed tallow FA or PHT). The calcium salts are made from palm
oil (higher in C16:0), soybean oil (higher in C18:2), or blend of fat sources.

Quality Factors
Rendered or processed fats originate primarily as recovered waste fats and can be highly variable in quality. The most important
measures of quality are solidification point (titre), saturation/unsaturation (usually measured by IV), total FA (TFA), free FA (FFA),
and moisture, insolubles, and unsaponifiables (MIU). Titre and IV both are estimators of unsaturation. Tallow is a triglyceride and
has a TFA content of 90% (10% is glycerol). Obviously, FFA should be 100% TFA. Total FA values less than 90% for triglycerides
indicate dilution with non-fat substances such as MIU. In many processed fats, the FFA value indicates the amount of “abuse” to
which the fat has been subjected, as heating and presence of water tend to hydrolyze or “split” triglycerides to FFA and glycerol.
However, livestock can utilize FFA, and their presence alone is a good indicator of fat quality since water has no energy value and
its presence dilutes the value of fat. Furthermore, water promotes splitting of fat, rancidity, and corrosion of storage tanks. Good
quality fats should contain no more than 1% moisture.

Feeding Practices
One of the original concepts with feeding fat was to reduce body weight (BW) loss during early lactation. However, research results
do not support this concept. The extra energy consumed by feeding fat in early lactation primarily supports higher milk yield. Since
the feeding of fat does not appear to reduce BW loss during early lactation and palatability problems sometimes exist with certain
fats, it is advised not to feed high levels of fat until 30 days postpartum. This strategy will allow the cow some time to adjust to the
lactational phase before fat is included in the diet; after all, the main strategy during the first 2 to 4 weeks of lactation should be to
maximize DM intake rather than energy intake. After intake has reached an acceptable level, then energy intake can be the focus.
There is some evidence that feeding fat may improve reproductive efficiency of dairy cows, independent of any effects on energy
balance of the cows (Lucy et al., 1991; Staples et al., 1998). Fat supplementation may increase the number and size of ovarian
follicles, increase plasma concentration of progesterone, reduce secretion of prostaglandin metabolites, and increase lifespan of the
corpus luteum (Staples et al., 1998). The FA profile of the supplemental fat is important for its positive impact on reproduction, with
linoleic acid being one of the causative FA. Therefore, this has lead to increased interest in feeding natural fat sources higher in
linoleic acid and the development of calcium salts higher in linoleic acid (Table 7.1).
The need for fat in diets should be based on the animals’ milk yield and body condition, quality of forages in diets (poorer quality
forages are lower in energy), and the level of DM intake by animals. As a guideline, cows can efficiently utilize as much dietary fat
as produced in milk, with appropriate adjustments for BW change (subtract amount lost or add amount gained in adipose tissue)
(Palmquist and Eastridge, 1991). The amount of fat that can actually be included in diets depends on the fat source, level of DM
intake, and fiber level in the diet. The importance of fat source in this regard was alluded to earlier. Cows consuming higher amounts
of DM and consuming diets adequate to high in fiber compared to diets marginal in fiber can handle a higher level of dietary fat.
Using the fiber equations by Jenkins (1997) are more applicable for typical diets and should be used with caution with low forage
diets based on high NDF from nonforage fiber sources.
Since DM intake influences the amount of fat to include in diets, fat levels to feed should be expressed as a percentage of DM
intake rather than on a weight basis. To provide supplemental fat, the natural fats should be used as a first priority because of their
lower cost. Generally speaking, oilseeds can be added to provide an additional 2% fat or tallow and animal-vegetable blends can be
used to provide 2.5 to 3% supplemental fat to diets (Table 1). The benefit of commercial fats become more apparent when total
dietary fat level must exceed 5% of dietary DM. Commercial fats are important for their rumen inertness and are convenient due to
their ease of handling. Price, availability, and characteristics that relate to palatability, inertness, and digestibility are important for
making comparisons among different commercial fats.
Insoluble soaps formed between fatty acids and cations, especially calcium and magnesium, in the lower portion of the small intestine
may reduce the absorption of calcium and magnesium (Jenkins and Palmquist, 1984; NRC, 2001). The evidence for this is equivocal,
but magnesium and calcium should be increased 20 to 25% in diets containing supplemental fat.

Effects of Supplemental Fat on Milk Composition

Adding fat to diets for lactating cows generally increases milk yield (if energy is limiting in the diet) and increases milk protein yield
but decreases milk protein concentration, typically by 0.1 to 0.2 percentage units. The metabolic processes attributing to this decline
in milk protein concentration has reviewed considerable attention, but the mechanism may still be uncertain (Schingoethe, 1996; Wu
and Huber, 1994).
Interest in the effects of supplemental fat on the FA composition of milk fat has increased and recently has been extensively
reviewed (Jensen, 2002). About 50% of the fat in milk is derived from de novo synthesis in the mammary gland (most of the 4:0 to
14:0 and about 50% of 16:0) and 50% from performed FA from either the diet or adipose tissue; however, the relative contribution
by preformed FA is higher during early lactation because of mobilization of adipose tissue. Supplemental fat also increases the
contribution of performed FA, thereby increasing the long-chain FA and decreasing the short-chain FA in milk. Supplemental dietary
fat may increase or have no impact on milk fat percentage, and feeding fat may decrease milk fat percentage if ruminal
fermentation is adversely affected, possibly related to the increased trans-18:1 in milk (Jensen, 2002).
Alteration of the FA composition of milk and the location of specific FA on glycerol can affect the processing properties of milk and
the nature (especially firmness) of dairy products (such as cheeses). Much interest continues in the concentration of conjugated
linoleic acid (CLA) in milk because of its anti-carcinogenic properties found in laboratory animals. Research continues with
attempting to increase CLA in milk by feeding different dietary sources of fat, altering biohydrogenation in the rumen, understanding
the role of delta-9 desaturase in the mammary gland, and understanding why cows on pasture have higher CLA in milk than cows
fed stored feeds.

Fat Analysis
Analysis of feeds for fat is not customary for many labs. Only high-fat feeds, by-product feeds, or blended feed mixtures containing
supplemental fat are worthy of fat analysis. Some by-product feeds are quite variable in fat, and thus, fat analysis would be advised.
Fat in feeds is usually analyzed either by ether extraction (EE) or by gas chromatographic methods. The EE procedure results in
higher values because it includes everything that is soluble in ether. The gas chromatographic methods provide a more precise
analysis because FA are actually measured. The FA content can be generally estimated from EE values by the following: forage EE
x 0.50; concentrate EE x 0.85, and tallow x 0.90. It is important to know which method is used so dietary levels of fat can be
consistent and accurate.

Conjugated linoleic acid

Dairy products and meat from ruminant animals are important sources of nutrients, supplying high quality protein, energy, and a
variety of minerals and vitamins. Research during the last few years reveals other nutritional benefits to the consumption of ruminant
food products, particularly dairy products. There is one compound in particular, conjugated linoleic acid (CLA), that excites
scientists, consumers, and producers, and may have far-reaching, positive effects on milk and meat consumption. CLA brings a
promising approach to redesigning food because milk has high levels of CLA, and CLA has been shown to have numerous potential
benefits for human health, including potent cancerfighting properties.

CLA Chemistry
All milk contains some fat (3.2 to 4.7%), but within milk there are a couple hundred different types of fat and fatty acids. The major
fatty acids in milk fat range from 4 to 20 carbon chain length. Linoleic acid, an essential dietary fatty acid, contains 18 carbons (C)
with two double bonds (C18:2) (see Figure 1-C). CLA is a term for specific isomers (forms) of linoleic acid with conjugated double
bonds (double bonds adjacent to each other C=C-C=C).
Of the 20 different isomers of CLA that have been identified, the cis 9-trans 11 (9c, 11t) form (commonly called “rumenic acid”) is
believed to be the most common natural form of CLA. The trans 10-cis 12 isomer has also been identified. Researchers are also
identifying and studying other potentially active isomers.

Fig 7.2: Isomers of CLA

CLA was first isolated and identified in the 80’s by Dr. Michael Pariza at the University of Wisconsin. Dr. Pariza isolated CLA
while looking for a compound in cooked hamburger that prevented skin cancer in mice. The cis 9, trans-11 isomer is the form most
commonly found in ruminant animals, and is the isomer thought to be most effective in lowering the risk of cancer in humans.

CLA Synthesis
Ruminal bacteria are key to the formation of CLA which explains why CLA production is unique to and found almost exclusively in
food products (milk and meat) produced from ruminant animals. CLA is an intermediate of the biohydrogenation of linoleic acid to
stearic acid. Biohydrogenation is a collective term used to describe the conversion of unsaturated to saturated fatty acids via
isomerization and hydrogenation of unsaturated fatty acids by rumen bacteria. This process is speculated to be a defense mechanism
by rumen bacteria against potentially toxic unsaturated fatty acids. During biohydrogenation of fatty acids, including the CLA
intermediate, fatty acids are continually leaving the rumen, being absorbed across the small intestine, and incorporated into milk fat.
The biohydrogenation and formation of CLA from linoleic acid has long been known as a source of CLA in the rumen. Another
major source of CLA is generated by other intermediates of biohydrogenation and sequential CLA synthesis in the mammary gland.
Biohydrogenation of oleic, linoleic, and linolenic acid all result in trans-11 (18:1). The trans-11 can be converted to 9c, 11t CLA in the
mammary gland by the tissue delta 9 desaturase enzyme. The importance of both of these sources has been studied, and mammary
synthesis may account for up to 70 to 80% of total CLA found in milk. However, it is important to keep in mind that the substrate for
mammary CLA synthesis originates in the rumen through microbial biohydrogenation. This demonstrates that the rumen is still a
primary source of CLA production.

Boosting concentrations of CLA in milk

Surveys have found an eight-to-ten fold variation in CLA concentrations in milk. This suggests that nutrition and herd management
practices may create a rumen environment that alters biohydrogenation of fatty acids and, in turn, the CLA concentrations in milk. In
Table 2 are listed the nutritional and ruminal factors that affect CLA concentration in milk fat.
Increases in CLA in milk were first noticed over 65 years ago when cows were turned onto spring pasture. The interest in CLA has
renewed the interest in increasing the CLA content of milk. A study at Penn State reported a two-fold increase in CLA with pasture
(5.4 to 10.9 mg/g of fat). This increase has been attributed to increased supply of fat substrate, and to potential changes in the rumen
environment and synthesis in the mammary gland. Adding supplements to the ration of grazing cows may diminish this effect.
Replacing conserved forages with fresh pasture clearly increases CLA concentrations in milk. A study comparing confinement
feeding of a TMR to pasture + TMR and pasture + concentrate clearly shows that feeding pasture elevated the CLA in milk. CLA
in the milk of cows fed a TMR was constant at 6 mg/g fat for the 18 week study. Cows fed pasture plus concentrate had elevated
CLA in milk by week 4 and 6, and the concentration peaked at 18 mg/g fat at week 18. Cows fed pasture + TMR had CLA
concentrations closer to that of cows fed TMR in confinement.
Fat supplementation and feed sources richer in unsaturated fatty acids have been shown to increase CLA in milk. Unsaturated plant
oils increase CLA more than feeding saturated animal fat sources. This is due to the lipid substrate available by the plant oils for
biohydrogenation to CLA and CLA precursors in the rumen. It follows then that increasing levels of plant oil and feeding calcium
salts of plant oils will increase levels of CLA in milk. Feeding plant seeds high in fat such as soybean, cottonseed, sunflower, and
flax (linseed) also increases CLA in milk.
However, if seeds are fed in a raw form, there is little change on CLA in milk. The seed coat must be broken making the fat
available to rumen biohydrogenation for CLA production. Supplementing with fish oil has been shown to increase CLA content of
milk when dairy cows on pasture were fed high oil seeds, the CLA content of milk increased more than when feeding pasture alone.
If we think back to the biosynthesis of CLA and the nutritional factors that increases CLA in milk, it is obvious there are many
pathways and nutritional factors that can increase CLA content of milk. Employing these nutritional factor together may increase
CLA content of milk beyond any one factor. A study was conducted to increase the levels of CLA in milk by affecting rumen
biohydrogenation and supplying lipid substrate. Cows were fed a TMR with the addition of corn oil, fish oil, or both. Including fish oil
has been shown to inhibit biohydrogenation allowing more intermediate products of biohydrogenation, including CLA and CLA
precursors, to escape the rumen and be incorporated into milk. Lipid substrates such as corn oil have shown to increase CLA
content of milk by providing more unsaturated fatty acids for biohydrogenation. When cows were fed both corn oil and fish oil in
combination, CLA content in milk increased ten-fold.

Potential health benefits

The potential benefit of CLA in human health is the major reason for the excitement and interest in CLA. The major interest
surrounding CLA is the anti carcinogenic or anti-cancer effects. Much of the research to date has been with laboratory animal
models. CLA can reduce new tumor growth and destroy existing tumor cells. CLA has killed existing cancer cells in colon, ovarian
and prostate carcinoma, leukemia, melanoma, and breast tumors. CLA enriched butter inhibited rat mammary tumor yield by 53%,
clearly showing the cis-9 trans-11 isomer was anticarcinogenic. In addition to the anti-carcinogenic properties, other positive health
benefits demonstrated in animal models include:
Reduced atherosclerosis
Enhanced immune system
Prevention and treatment of diabetes
Weight reduction; reduced body fat and increase body protein
Enhanced bone formation
As research continues on many fronts, the specific physiological effects and the responses will be better defined.

Importance of dietary CLA in pig feeding

Concerns about the development of antibiotic-resistant bacteria have led to feeding bans on several antibiotics in the European
Union. One alternative to antibiotics may be the incorporation of conjugated linoleic acid (CLA) into swine diets. Research indicates
that CLA can modify the immune system of weanling pigs by altering the type and number of immunocytes (Bassaganya- Riera et
al., 2001). Pigs fed diets supplemented with CLA have greater gain: feed efficiency and leaner carcasses than pigs not fed CLA
(Dugan et al., 1997; Eggert et al., 1999). A portion of the increase in efficiency and lean growth could perhaps be attributed to
CLA’s modulation of the immune response in swine (Bassaganya-Riera et al., 2001). However, researches on the effect of CLA
on the overall performance of pigs are very limited.

Conclusion
Fats are very useful for increasing energy density of diets for high-producing dairy cows. Similar to other feeding changes, fat should
be gradually introduced into diets. Physical and chemical properties of available fat sources, animal’s milk yield, body condition, and
level of DM intake, and associated costs are factors for consideration when feeding fat to dairy cows. Use of feed grade fats on
dairy farms is expected to continue because of increasing milk yield per cow. Different commercial or specialty fats will continue to
be available for feeding upper levels of fats in diets and for targeted roles based on new research. The presence of a compound
(CLA) in ruminant fat with such potent health promoting effects has been an unanticipated discovery. The ability to enhance the
concentration of CLA through manipulation of the dairy ration demonstrates the feasibility of producing CLA enriched dairy
products. As consumers become more conscious of the link between diet and health, milk designed to have enhanced levels of CLA
may provide new market opportunities for milk and milk products such as butter and cheese.
Table 7.1. Fatty acid composition and characteristics of different fat sources

Fat Source Fatty acids (weight %) Fat C18 IV Fat diges

14:0 16:0 16:1 18:0 18:1 18:2 18:3 (EE) Unsat.FA (%) tibility (%)
(%)
Oil seeds
Cotton seed 0.08 22.7 0.8 2.3 17.0 51.5 0,2 19.3 68.7 107 86
Linseed oil — 5.3 — 4.1 20.2 12.7 53.3 100 86.2 185 86
cake
Soybeans 0.1 10.3 0.2 3.8 22.8 51.0 6.8 19.0 80.6 131 86
Sunflower — 5.4 0.2 3.5 45.3 39.8 0.2 41.9 85.3 113 86
Animal fats
Tallow 3.0 24.5 3.7 19.3 40.9 3.2 0.7 99.8 44.8 48 68
Yellow 1.8 22.1 3.5 11.5 43.7 14.6 0.9 99.0 59.2 72 —
grease
Commercial
fats
Calcium salt 1.3 48.6 1.1 4.1 36.5 7.8 0.3 84.5 44.6 49 86
Megalac — 47.1 — 4.7 36.5 9.4 1.2 85.0 47.1 53.0 —
Rumolac — 9.0 — 9.0 31.5 38.5 — 82.9 70.0 98.0 —
Table 7.2. Factors affecting CLA Concentrations in Milk Fat

Factors Effect on CLA concentration in milk fat

Forages
Pasture versus TMR1 Increased with consumption of pasture
Forage: concentrate ratio2 Increased with high forage diet
Maturity of forage1 Increased with less mature forage
Plant oils
Unsaturated versus saturated3 Increased with the addition of unsaturated oils
Increased by feeding higher levels
Amount of plant oils3
Calcium salts of plant oils3 Increased with increasing amounts
Plant seeds
Raw seeds No Effect
Processed seeds3 Increased over raw seeds
Other
Plant oil versus animal fats3 Increased with plant oils
Fish oil2 Increased in relation to level fed in the diet
1CLA increases may be due to both addition of lipid substrate and modified rumen environment
2CLA increases may be due to a modified rumen environment
3CLA
increases may be due to addition of a lipid substrate

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 8 Phytobiotics

Phytobiotics are plant derivatives such as herbs, plant extracts or spices and have a wide range of activities such as stimulation of
feed intake, growth and endogenous secretions in the gut. They act as immunomodulators resulting in decreased mortality and also
have coccidiostatic, anti microbial, anthelminthic and anti-inflammatory activities. Phytobiotics also possess hepatoprotective and
hepatogenic properties, which tone up liver resulting in increased nutrient utilization and better performance. Herbs like Achyranthes
aspera (Prickly Chaff Flower, Devil’s Horsewhip, Apamarga), Andrographis paniculata (Green chirayta, King of bitters,
Kalamegha), Azadirachta indica (Neem), Boerhaavia diffusa (Spreading Hogweed, Punarnava), Eclipta alba (False Daisy,
Bhringaraj), Ichnocarpus frutescens (Black creeper, Utpalagopa), Terminalia chebula (Black myrobalan, Haritaki). These herbs
have hepato-stimulant, hepato-protective, immunomodulatory and antioxidant activities (Sadekar et al., 1998; Manu and Kuttan,
2009; Michels et al., 2011, Dash et al., 2007). Further, they optimize digestion and metabolism resulting in better protein utilization,
improved mucosal function and reduced cost of metabolic deamination. Andrographolide and 14-deoxy-11, 12-didehydro-
andrographolide isolated from Andrographis paniculata inhibits free radical activities and lipid peroxidation. Inhibition of lipid
peroxidation in meat prevents free radical production thereby preserving meat composition, colour and improvement in shelf life. In
addition, it has been studied that a herb Terminalia chebula helps to reduce stress (Selvakumar et al., 2007).
Use of herbs for treatment of animals and human beings has been from very ancient times. Herbal medicine has its origins in ancient
cultures including those of the Egyptians, American, Indians and Chinese. Varieties of plants of medical importance were used by
man even during the Treta Yuga. The earliest compilation of drugs and illness appeared in “AYURVEDA” which is claimed to have
been written around 3000 BC. However, the concept gained momentum in the scientific world only in the later part of last century.
Various herbs or herbal products are used for treatment of different diseases. It has been estimated that around 8000 different types
of natural medicines are in use in China. Most of the natural medicines are made of different plant products.
The herbs and their extracts increase appetite, stimulate digestive enzymes, modulate immune system and have anti-helminthic and
anti-coccidial effect. In recent years, several studies have been carried out to test the efficacy of different herbs and their secondary
metabolites on the growth, immunocompetence and production traits of livestock and poultry. However, it is beyond the scope of this
chapter to discuss all of them. Hence, an attempt has been made to discuss the role of some commonly used herbs and plant
secondary metabolites in livestock and poultry.

Curcuma longa (Haldi)

Turmeric has a long tradition of use in the Chinese and Ayurvedic systems of medicine, particularly as an anti-inflammatory agent,
and for the treatment of flatulence, jaundice, menstrual difficulties, hematuria, hemorrhage and colic. Turmeric is known as
stomachic, blood purifier, and is useful in many conditions like common cold, leprosy intermittent fever, infections of liver, dropsy,
purulent ophthalmia, otorrhea, pyogenic infections, wound healing and infections etc.
Turmeric can also be applied topically to relieve pain and inflammation. Current research has focused on turmeric’s antioxidant,
hepatoprotective, anti-inflammatory, anticarcinogenic, and antimicrobial properties, in addition to its use in cardiovascular disease and
gastrointestinal disorders. Some of the studies on turmeric by various workers have been mentioned in this section.
Soni et al. (1992) studied the effect of extracts of turmeric (Curcuma longa), Garlic (Allium sativum) and Asafetida (Ferula
asafetida) on aflatoxin production by Aspergillus parasiticus in vitro and found 90% inhibition of aflatoxin at concentration of 5-
10mg/ml. They also reported the reverse changes in damaged liver, fatty change, necrosis and bilary hyperplasia by these feed
additives. Kurkure et al. (2000) concluded that turmeric treatment can only partially ameliorate low grade aflatoxicosis in chicks.
Sultan (2003) reported that turmeric (feed additive) at a level of 0.5% enhances overall performance of broiler chickens.
Emadi and Kermanshahi (2006) observed that addition of turmeric rhizome powder (TRP) into the diets significantly decreased
relative abdominal fat pad and heart weights to live body weight. So they concluded that TRP may improve carcass quality and
produce leaner meat. Further, they pointed out that use of TRP as a reducing heart weight factor, may show some improvement in
circulatory and respiratory systems.

Terminalia arjuna
Arjuna tree is found throughout the sub-Himalayan tracts, the Deccan region, Srilanka and the south eastern countries. The bark is
used to heal wounds. It is also used in heart-disease, contusions, and fractures. Juice of leaves is used in ear-aches. It is one of the
best herbs for heart disease (prevents and helps in the recovery of) angina and heals heart tissue scars after surgery. Also useful for
bile, edema, fractures, broken bones, diarrhea malabsorption and venereal disease as well as external treatment for ulcers, acne, skin
disorders. Arjuna bark is used in the form of decoction (1 in 10) in doses of half to one ounce in hemorrhages. Also, it is used in
diarrhea, dysentery and sprue. It is also useful in bilious affections, and as an antidote to poisons. It is used as a remedy for scorpion
sting.
In recent years, lot of studies has been carried out to study the medicinal properties of different parts of T. Arjuna. Some of these
studies have been briefly discussed.
Khanna et al. (1996) observed that supplementation of Terminalia arjuna bark powder (100 mg/kg diet) to hyperlipaemic rats
lowered serum lipids and increased HDL cholesterol levels. Sivalokanathan et al. (2006) studied on ethanolic extract of arjuna bark
on carbohydrate metabolizing enzymes of N-nitrosodiethylamine induced hepatocellular carcinoma in Wistar albino rats. The plasma
and liver glycolytic enzymes such as hexokinase, phosphoglucoisomerase, aldolase were significantly increased in cancer induced
animals while glyconeogenic enzyme; glucose-6-phosphatase was decreased. These enzymes were reverted significantly to near
normal range in treated animals after oral administration of T. arjuna for 28 days. The modulation of the enzymes causing depletion
of energy metabolism leads to inhibition of cancer growth. This inhibitory activity may be due to the anticancer activity of
constituents present in the ethanolic extract of T. arjuna.
Raghavan and Kumari (2006) reported that ethanolic extract (250 and 500 mg/kg body weight) of Terminalia arjuna stem bark in
alloxan induced diabetic rats produced significant reduction in lipid peroxidation. The effect of oral T. arjuna at the dose of 500
mg/kg body weight was more than the 250 mg/kg body weight. The extract also causes a significant increase in superoxide
dismutase, catalase, glutathione peroxidase, glutathione-s-transferase glutathione reductase and glucose-6-phosphate dehydrogenase,
reduced glutathione, vitamin A, vitamin C, vitamin E, total sulfhydryl groups and non protein sulfhydryl groups in liver and kidney of
alloxan induced diabetic rats, which clearly shows, the antioxidant property of T. arjuna bark. The result indicates that the extract
exhibits the antioxidant activity through correction of oxidative stress and validates the traditional use of this plant in diabetic animals.

Emblica officinalis (Amla)

The fruits of Emblica officinalis are sour, astringent, bitter, acrid, sweet, cooling, anodyne, ophthalmic, carminative, digestive,
stomachic, laxative, aphrodisiac, rejuvenative, diuretic, antipyretic and tonic. They are useful in vitiated conditions of diabetes, cough,
asthma, bronchitis, cephalalgia, ophthalmopathy, dyspepsia, colic, flatulence, hyperacidity, peptic ulcer, erysipelas, skin diseases,
leprosy, haematogenesis, inflammations, anemia, emaciation, hepatopathy, jaundice, diarrhoea, dysentery, hemorrhages, leucorrhoea,
menorrhagia, cardiac disorders, intermittent fevers and greyness of hair. Some of the studies conducted on E. officinalis have been
mentioned below.
Jain and Khurdiya (2004) reported Indian gooseberry juice contains the highest vitamin C (478.56 mg/100 ml). Bhattacharya et al.
(1999) reported that the antioxidant activity of E. officinalis may reside in the tannoids of the fruits of the plant, which have vitamin
C like properties, rather than vitamin C itself. Rege et al. (1999) reported Emblica officinalis strengthened the defence
mechanisms against free radical damage induced during stress. The effect of Emblica officinalis appeared to depend on the ability
of target tissues to synthesize prostaglandins.
Bhattacharya et al. (2000) concluded that anti-stress activity of E. officinalis may be at least partly due to its tendency to normalize
stress-induced perturbations in oxidative free radical scavenging activity as several stress-induced diseases, including the process of
aging, may be related to accumulation of oxidative free radicals in different tissues. Nosal’ova et al. (2003) reported that antitussive
activity of the dry extract of Emblica officinalis is not only due to antispasmolytic and antioxidant efficacy effects but also due to its
effect on mucus secretion in the airways. Panda and Kar (2003) reported that the ethanolic extract from the fruits of Emblica
officinalis may potentially ameliorate the hyperthyroidism with an additional hepatoprotective benefit.
Rajak et al. (2004) reported chronic Emblica officinalis administration causes myocardial adaptation by augmenting endogenous
antioxidants and protects rat hearts from oxidative stress associated with ischemic-reperfusion injury. Rao et al. (2005) reported the
oral administration of amla extracts to the diabetic rats slightly improved body weight gain and also significantly alleviated various
oxidative stress indices of the serum of the diabetic rats. In addition, the decreased albumin levels in the diabetic rats were
significantly improved with amla. These results form the scientific basis supporting the efficacy of amla for relieving the oxidative
stress and improving glucose metabolism in diabetes. Sancheti et al. (2005) reported Emblica officinalis (Family: Euphorbiaceae)
indigenous to India, is valued for its unique tannins and flavonoids, which contain very powerful antioxidant properties. Scartezzini et
al. (2006) reported Emblica fruit contains ascorbic acid (0.4%, w/w), and that the ayurvedic method of processing increases the
healthy characteristics of the fruit due to a higher antioxidant activity and a higher content of ascorbic acid (1.28%, w/w). It has also
been found that Vitamin C accounts for approximately 45-70% of the antioxidant activity.

Azadirachta indica (Neem)

Neem is very effective against any kind of infection occurring in body. It is also regarded as herbal antibiotic. It helps in healing of
wounds, keeps wounds unaffected by infections, reduces inflammation and reduces pus formation. It also helps in preventing itching.
It is helpful in worm infestation. It is also helpful in relieving from burning sensation in acidity. It helps in purifying blood and is useful
in diabetes and urine related problems. It also helps in relieving from respiratory problems as it is mucolytic and expels out extra
mucus present in the respiratory tract. It is very effective in skin related ailments, it also promotes digestion. It is very effective in
fever especially caused in malaria. It improves eyesight. It removes foul odor from mouth and strengthens gums and teeth.
The bark is bitter, astringent, acrid, refrigerant, depurative, antiperiodic, vulnerary, demulcent, insecticidal, liver tonic, expectorant,
urinary astringent, anthelmintic, pectoral and tonic. It is useful in vitiated conditions of pitta, hyperdipsia, leprosy, skin diseases,
eczema, leucoderma, pruritus, intermittent and malarial fevers, wounds, ulcers, burning sensation, tumour, tubercular glands,
anorexia, vomiting, dyspepsia, intestinal worms, hepatopathy, cough, bronchitis, urorrhea, diabetes, inflammation, amenorrhea,
lumbago, haemorrhoids, otalgia, syphilis and fatigue. The leaves are bitter, astringent, acrid, depurative, antiseptic, ophthalmic,
anthelmintic, alexeteric, appetiser, insecticidal, demulcent and refrigerant. They are useful in vitiated conditions of pitta, burning
sensation, leprosy, skin diseases, leucoderma, pruritus, ophthalmopathy, intestinal worms, dyspepsia, ulcers, tuberculosis, boils,
eczema and malarial and intermittent fevers. Some of the studies done on poultry is mentioned below.
Uko and Kamalu (2005) observed when raw and autoclaved neem seed kernels were ground and incorporated into standard basal
diet at 150 and 225g/kg as substitutes for ground nut cake, there was lymphocytosis and higher levels of neem seed kernels
decreased alkaline phosphatase activity in cockerel chicks. In another experiment, Uko and Kamalu (2006) observed when full fat
neem seed kernel was incorporated in the diet of cockerel chicks, eviscerated carcass weight decreased and yield increased
significantly in cockerel chicks with increased levels of neem kernel. Uko et al. (2006) observed when raw full fat neem seed kernel
replaced peanut at graded doses in the diet of cockerel chicks, aspartate aminotransferase activity was elevated while cholesterol
concentration was lowered. However, activity of alanine aminotransferase and concentrations of bilirubin, creatinine and uric acid
did not differ significantly among the cockerels. Further, they observed emaciation and paleness of carcasses, kidney congestion and
enteritis.
Lather et al. (2002) observed that feeding of neem seed cake to broiler chickens resulted in lower alanine aminotransferase and
higher aspartate amino transferase values. However they observed no significant changes in the concentration of uric acid,
creatinine, sodium and potassium with feeding of neem seed cake to broiler chickens.

Ocimum sanctum (Tulsi)

Tulsi (Ocimum sanctum) contains eugenol, carvacrol, methylchavicol. Tulsi contains a volatile oil consisting of about 70% eugenols
as well as methyl eugenol and caryophyllene. Other constituents with likely pharmacological activity include the triterpenoidursonic
acid, rosmarinic acid, alkalids, saponins, flavonoids (including apigenin and luteolin and glycosides thereof), phenylpropane glucosides
and tannins. The seeds of tulsi contain affixed oil containing five fatty acids, including about 17% linolenic acid and just over 50%
linoleic acid.
Tulsi has a long history of medicinal use, and is mentioned in ancient Ayurvedic text such as Charak Samhita. Tulsi is used for
treatment of variety of conditions including pain, fever, vomiting, bronchitis, earache and diseases of heart and blood. It is also used
in case of diabetes, arthritis and asthma. Fresh leaves taken with black pepper used as a prophylactic against malaria and a
decoction of the root are recommended for malarial fever. The leaf juice is used in chronic fever, hemorrhage, dysentery and
dyspepsia. It is used also as an anthelmentic and is topically used for ringworm and skin diseases. Its pounded leaves are mixed with
sandalwood and applied topically for relief in headache. Tulsi seeds are used in anti leucoderma preparations. Zheljazkov et al.
reported that essential oil content of O.bacilium had in vitro activity against Leishmania donovani. Some of the studies in tulsi
pertaining to poultry have been mentioned below.
Gupta and Charan (2007) studied the non toxic doses of Ocimum sanctum in broiler chicken. It was found that maximum weight
gain was observed in the group of chickens treated with 200 mg of dried leaves powder per birds daily for 15 days as compared to
control group. Hindustani and Singh (2006) reported that Zeetress, a herbal preparation containing extract of Ocimum sanctum and
Withania somnifera as major ingredient added to the drinking water @ 5mg per bird from day 3 to 28 and 10mg/bird from day 29 to
42 had immunopotentiating effect in vaccinated chicken with better immune response to Newcastle disease virus vaccine, lower
lesion scores, improvement in live weight gain and feed conversion efficiency. Rao et al. (1999) studied the effect of Zeetress, a
polyherbal preparation containing Withania somnifera, Ocimum sanctum, E. officinalis on chicken and found that administration of
zeetress minimized stress and had a positive effect on weight gain, feed conversion, general resistance and livability. Babu et al.
(2002) studied the effect of zeetress, a poly herbal compound containing (Ocimum sanctum, Withania somnifera and Emblica
officinalis) against cypermethrin induced immunosuppression and reported that cypermethrin significantly reduced the phagocytic,
opsonic indices and body weight, where as zeetress significantly restored the affected phagocytic , opsonic indices and body weight.
Batra et al. (2004) studied the clinicopathological and immunological effect of Ocimum sanctum in chicken. Tulsi leaf powder
supplemented @ 5 gm/kg feed did not cause any pathological lesion in the tissue. Delayed type hypersensitivity response against di
nitro chloro benzene antigen was significantly increased in the Ocimum sanctum leaf powder group, indicating the immuno
modulatory activity of tulsi leaf. Lanjewar et al. (2008) reported that supplementation of tulsi leaf powder @ 0.5% and 1.0%
significantly increased the final body weight and weekly body weight gain in broiler chicken. Feed conversion ratio also improved in
1.0% tulsi leaf powder group. They also noted that serum HDL cholesterol level was significantly increased with supplementation of
tulsi in diet. The net profit found was more in birds supplemented with 0.5% Tulsi leaf powder than 1.0% Tulsi leaf powder. They
concluded that supplementation of tulsi leaf powder @ 1.0% in broiler diet for 42 days reduced the meat and serum cholesterol with
increase in serum HDL–cholesterol. Mamta and Mishra (2006) reported that administration of tulsi dry leaf powder @ 5 g/kg of
feed enhanced cell mediated immune response as observed by DTH in terms of increased foot pad thickness to sonicated bacterial
cell protein (SBCP) antigen. It also enhanced humoral immune response against S. pullorum plain antigen as detected by standard
tube agglutination (SAT) test. The present findings indicate that tulsi dry leaf powder has an immunopotentiating effect.

Plant secondary metabolites

Reduced social acceptance of antibiotics in animal feeds leads the search for alternatives, and plants extracts are attractive with
consumer opinion that most things ‘natural’ are good. Plants produce an extensive variety of organic compounds derived from their
secondary metabolism that are classified in three main groups: saponins, tannins, and essential oils.

Saponins
Effects on animal growth and feed intake
Animal nutritionists have generally considered saponins to be deleterious compounds. In ruminants and other domestic animals the
dietary saponins have significant effects on all phases of metabolism, from the ingestion of feed to the excretion of wastes (Cheeke,
1996). Lucerne and soybeans are the main examples of saponin-rich plants that serve extensively in human, ruminant and poultry
diets. Recently, a number of studies have reported both beneficial and adverse effects of these compounds in a variety of animals
(Sen et al., 1998).
Y. schidigera plant extract (saponin containing plant) has been found to improve growth, feed efficiency and health in ruminants
(Mader & Brumm, 1987). Quillaja saponins increased the efficiency of in vitro rumen-microbial protein synthesis and decreased
degradability of feed protein (Makkar and Becker, 1996). Partially hydrolysed lucerne saponins administered intra-ruminally resulted
in a significant reduction in the total protozoa count in the rumen of sheep (Lu and Jorgensen, 1987) which may be the reason for the
decrease in feed protein degradability. Saponins are considered to have detrimental effects on protozoa through their binding with
sterols present on the protozoal surface. Sterols are absent on bacterial membranes. Yucca extract can also bind NH 4 when ruminal
NH4 concentrations are high, and release it again when ruminal NH4 is low, providing a continuous and adequate supply of NH 4 for
microbial protein synthesis (Hussain & Cheeke, 1995). Supplementation of feed with leaves of Sesbania sesban, known for its high
saponin content, has been found to have the potential to improve protein flow from the rumen by suppressing protozoal action there
(Newbold et al., 1997) but rumen bacteria were observed to be capable of metabolising the antiprotozoal factor. The positive effects
of saponins were more pronounced when they were directly administered into the rumen rather than added to the feed (Odenyo et
al., 1997). Killeen et al. (1998) proposed that a surfactant or flocculent action of saponins on the feed constituents that alters the
rate of digestion would account for the substrate-dependent nature of the effect of Y. schidigera on rumen DM and N digestibility.

Effects on the immune system

Saponin-based adjuvants have the unique ability to stimulate the cell-mediated immune system, as well as to enhance antibody
production, and have the advantage that only a low dose is needed for adjuvant activity (Oda et al., 2000). There have been quite a
few reviews on the immunostimulatory activity of saponins (Barr et al., 1998; Sjolander et al., 1998). Research thus far has
concentrated on Quillaja saponins and derivatives such as immune-stimulating complex vaccines (immunestimulating complexes
formed by the combination of cholesterol, saponin, phospholipid and amphiphatic proteins). Quillaja and other saponins either as
crude mixtures or as purified compounds have been reported to increase immune-cell proliferation in vitro (Plohmann et al., 1997;
Lacaille-Dubois et al., 1999). Purified Quillaja saponins boosted antibody production without producing any reaginic antibodies (So et
al., 1997). Immune-stimulating complexes formulated with Quillaja saponin preparations induced specific cytotoxic T-lymphocyte
responses (Coulter et al., 1998) and have been reported to induce antibody responses and/or protective immunity in guinea-pig,
turkey, cat, rabbit, dog, seal, sheep, pig, cow, horse and monkeys (Mowat et al., 1999). The adjuvant action of saponins was,
however, not so pronounced in some of the non-mammalian species tested.
The mechanisms of immune-stimulating action of saponins have not been clearly understood, but many explanations have been put
forward. Saponins reportedly induced production of cytokines such as interleukins and interferons that might mediate their
immunostimulant effects (Jie et al., 1984). It is likely that they interact with antigen-presenting cells to induce many of these
responses (Barr et al., 1998). The incorporation of the saponins into cell or endosomal membranes might expose the incorporated
antigen to cytosolic proteases. As against the stimulatory effects on specific immunity components, saponins have also been shown
to be able to prevent some non-specific immune reactions such as inflammation (Haridas et al., 2001) and monocyte proliferation
(Delmas et al. 2000; Yui et al. 2001). One downstream effector substance whose activity has shown to be lowered by saponin is
phospholipase A2 (de Oliveira et al., 2001) that causes a decrease in hydrolysis in membrane phospholipids and thereby decreased
membrane fluidity. The varied action of saponins might indicate that the immunostimulatory effects of saponins are the result of
specific targeting of physiological intermediaries rather than the result of a non-specific effect on cell membrane permeability.

Antioxidant effects
The importance of the antioxidants contained in foods is well appreciated for both preserving the foods themselves and supplying
essential antioxidants in vivo. However, the term ‘antioxidant’ is very loosely used. Often, the term is used to describe chain-
breaking inhibitors of lipid peroxidation as free radicals generated in vivo damage many targets other than lipids, including proteins,
DNA and small molecules. These oxidation reactions might lead to an array of adverse biological effects. Some of the protection
mechanisms afforded by saponins against this have already been discussed earlier. Zilversmit (1979) hypothesised that atherogenesis
might result from phenomena that occur immediately after eating, and that it might be affected by chylomicron remnants. Several
researchers (Staprans et al., 1994; Wolff and Nourooz-Zadeh, 1996; Ursini et al., 1998) expanded Zilversmit’s hypothesis and
suggested that dietary lipid hydroperoxides, which may be partly generated during digestion in the alkaline pH of the intestine, are the
source of chylomicron remnants of lipid hydroperoxides, which are elevated in the postprandial state. These findings emphasise the
importance of natural antioxidants in food, and throughout the digestive tract. Usually, polyphenols and carotenoid pigments, being
the major nutritional antioxidants in food, attract most of the research in this area. Some saponins have also been found to have
antioxidative or reductive activity.
Saponin mixtures present in plants and plant products possess diverse biological effects when present in the animal body. With the
information presently available, it is difficult to establish clear functionality and structure– activity relationships regarding the effects
of saponins in biological systems. This is largely due to the occurrence of a vast number of saponins with similar chemical structures,
and to the complexity of cellular physiological reactions, which are often differently influenced by small and subtle differences in
stereo-structures of effector ligands. Other factors that could have substantial influence on saponin actions could be their
interactions with other dietary constituents.

Tannins
Definition and classification
The tannins are a group of plant secondary compounds which have been known and used by man for centuries. Their name comes
from the French tan meaning the bark of the holm oak and other trees used in tanning. From a chemical point of view it is difficult to
define tannins since the term encompasses some very diverse oligomers and polymers (Harborne, 1999; Schofield et al., 2001). It
might be said that the tannins are a heterogeneous group of high molecular weight phenolic compounds with the capacity to form
reversible and irreversible complexes with proteins (mainly), polysaccharides (cellulose, hemicellulose, pectin, etc.), alkaloids, nucleic
acids and minerals, etc. (Giner-Chavez, 1996; Schofield et al., 2001). The tannins have traditionally been divided into two groups: the
condensed and the hydrolysable tannins. Hydrolysable tannins (HT) are made up of a carbohydrate core whose hydroxyl groups are
esterified with phenolic acids (mainly gallic and hexahydroxydiphenic acid). The condensed tannins (CT), or proanthocyanidins, are
non-branched polymers of flavonoids units (flavan-3-ol, flavan-3,4-diol), and usually have a higher molecular weight than the HT
(1000-20000 Da compared to 500-3000 Da) (McLeod, 1974; Mueller- Harvey and McAllan, 1992; Mueller-Harvey, 1999). Although
this division of the tannins is the most widely accepted, many authors believe it does not fully reflect their chemical complexity
(Mueller-Harvey and McAllan, 1992; Mueller-Harvey, 1999).

The distribution of tannins in nature

The tannins are widely distributed throughout the plant kingdom, especially among trees, shrubs and herbaceous leguminous plants
(McLeod, 1974). The range of species over which these compounds are found has grown as detection techniques have improved.
Despite the general idea that tannins are only found in plant species from tropical or arid/semi-arid areas (Giner-Chavez, 1996), they
are found in those of other regions. In general, tannins are more abundant in the parts of the plant that are most valuable to it, e.g.,
new leaves and flowers (which are more likely to be eaten by herbivores) (Terril et al., 1992; Van Soest, 1994). Numerous reports
illustrate the effects of environmental and seasonal factors as well as of phenological development. Very briefly, high temperatures,
water stress, extreme light intensities and poor soil quality increase the tannin content of plants (Van Soest, 1994).

Effect of tannins in ruminant nutrition

Tannins can be beneficial or detrimental to ruminants, depending on which (and how much) is consumed, the compound’s structure
and molecular weight, and on the physiology of the consuming species (Hagerman and Butler, 1991). It is important to remember
that all the quantities mentioned in this revision should be taken with great caution since different analytical methods and especially
different standards (e.g., quebracho, tannic acid, catequin, cyanidin, delphinidin or internal standards from the plant itself etc.) can
provide very different —and therefore ambiguous— results (Giner-Chavez, 1996; Schofield et al., 2001).

Practical use of tannins Treatments to protect dietary protein from ruminal degradation
One of the basic goals of protein nutrition in ruminants is to optimise dietary protein use in order to maximize animal growth and milk
production per unit of protein consumed (Schwab, 1995). As mentioned several times, tannins could protect dietary proteins from
ruminal degradation. With respect to HT, Driedger and Halfield (1972) managed to reduce the in vitro ruminal protein degradability
of soybean meal through treatment with tannic acid. Its effect on intestinal digestibility however, was not very consistent. Terril et
al. (1992) observed that the CT of quebracho provoked a greater reduction in the degradability of soybean meal than commercial
tannic acid, but in general the results obtained were very variable and depended on many factors. One of the drawbacks of using
tannins as additives to protect protein rich feeds is the possibility of their degradation by rumen microorganisms. If this were to
happen, the treated feeds would be just as vulnerable to ruminal degradation as untreated feeds. However, more research is needed
in this area since HT are easily hydrolysed and their effect could easily be nullified by the rumen microbiota.
Although somewhat obvious, it is worth pointing out that proper management of natural tannin-containing resources (e.g., selective
grazing or supplementing the diet with the right kind of shrubs) could provide the same beneficial effects with respect to protein
degradation.

Bloat prevention
It is well documented that bloat occurs when grazing ruminants consume large quantities of leguminous plants (e.g., alfalfa or
clover). The gases produced in the rumen during fermentation cannot be released in the normal way since they are trapped in a
persistent foam caused by the rapid release of soluble proteins during chewing and ruminal degradation. However, when these
animals graze on leguminous plants containing CT (for example Onobrychis viciifolia) this does not occur (McMahon et al., 2000).
The substitution of a small amount (approximately 10%) of ingested alfalfa DM by Onobrychis viciifolia provides unquestionable
benefits in the prevention of bloat (McMahon et al., 1999 and 2000). The problem of this strategy is, however, the low persistence
of this plant species in mixed cropping with alfalfa. The possibility of genetically modifying alfalfa to produce CT has been suggested
on several occasions and has been the subject of several studies (McMahon et al., 2000). However, the difficulty of the molecular
techniques required has made progress slow. Very recently, the preliminary results of a study on the ruminal fermentation of
transgenic alfalfa were published. The Lc gene of maize was introduced into alfalfa to induce the synthesis of CT (Wang et al.,
2003). The modification of the alfalfa decreased its initial rate of degradation in the rumen, but not the extent of degradation. This
offers an interesting way to help to prevent bloat.

Control of internal parasites

The tannins of numerous plant species help to control certain internal parasites of animals, for example the nematode
Trichostrongylus colubriformis (Butter et al., 2000). It is speculated that the positive effect on the host animal might be associated
with a direct negative effect on the parasites themselves plus an indirect effect in the form of increased availability and digestive
utilization of protein (Min and Hart, 2003). The literature has several examples of this in sheep and goats grazing L. corniculatus or
Hedysarum coronarium (Robertson et al., 1995) and after having ingested quebracho CT (Butter et al., 2000) etc.

Essential oils
Essential oils (EO) are blends of secondary metabolites obtained from the plant volatile fraction by steam distillation (Gershenzon
and Croteau, 1991). The term “essential” derives from “essence,” which means smell or taste, and relates to the property of these
substances of providing specific flavors and odors to many plants. They are characterized as having a very diverse composition,
nature, and activities. The most important active compounds are included in two chemical groups: terpenoids (monoterpenoids and
sesquiterpenoids) and phenylpropanoids.
Few studies have been published on effects of EO or their constituents on milk production and composition of dairy cows. Benchaar
et al. (2006b, 2007) observed no changes in DM intake, milk production, and milk components when dairy cows were fed 750 mg or
2 g of MEO daily. Similarly, supplementation of dairy cows with peppermint at 20 g/kg DM had no effect on milk yield and milk
composition (Hosoda et al., 2005). More recently, Yang et al. (2006) observed that addition of garlic (Allium sativa, 5 g/day) and
juniper berry (Juniperus communis, 2 g/day) oils to dairy cow diets had no effect on DM intake, milk production or milk
composition. In these studies, the lack of effect of EO and their active components on milk performance was consistent with the
absence of effects of these plant extracts on feed intake and ruminal fermentation.
EOs have an antibacterial activity against Gram-negative and Gram-positive bacteria (Helander et al., 1998). Several Gram-positive
bacteria are involved in ruminal biohydrogenation of unsaturated dietary fatty acids (Harfoot and Hazlewood, 1988). Therefore,
feeding EO could lower biohydrogenation of fatty acids by reducing the number, and the activity, of bacteria involved in the
biohydrogenation of unsaturated fatty acids. Benchaar et al. (2007) reported no change in milk fatty acid profile when cows were
supplemented daily with 750 mg of MEO. However, supplementing the same mixture at a higher concentration (i.e., 2 g/day)
increased the concentration of conjugated linoleic acid (CLA), a health-promoting fatty acid, in milk fat.
Data on effects of EO and their compounds on beef cattle performance are almost nonexistent. In one study, Benchaar et al.
(2006a) evaluated growth performance of beef cattle fed a silage base diet supplemented with 2 or 4 g/day of a commercial mixture
of EO compounds (Vertan®, IDENA, Sautron, France) consisting of thymol, eugenol, vanillin and limonene. Results showed that
DM intake and average daily gain were not affected by the addition of this EO compounds mixture. However, the gain to DM intake
ratio was affected quadratically with a dose of 2 g/day maximizing feed efficiency.

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