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CENTRAL INSTITUTE OF FISHERIES EDUCATION (Deemed
University) Indian Council of Agricultural
Research Panch Marg, Off Yari Road, Mumbai 400
061
Antinutritional factors and its effects on
Immune system
Dr K Pani Prasad Principal Scientist Department
of Aquatic Animal Health Management

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Introduction

• Anti-nutritional factors (ANF) are compounds
  which act to reduce nutrient utilization and or
  food intake (Osagie, 1998).
• These antinutritional factors play a great role
  in limiting the wider use of many plants.
• Natural compounds capable of precipitating
  deleterious effects in man and animals (Osagie,
  1998).

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• The levels of toxic substances in plants vary
  with the species of plant, cultivar and
  post-harvest treatment such as soaking, drying,
  autoclaving and seed germination.
• These anti-nutritional factors are also known as
  secondary metabolites in plants and they have
  been shown to be highly biologically active
  (Zank, 1991).
• Antinutritional factors can have adverse effects
  over the health of the consuming organism.
• Consuming improperly processed foods especially
  legumes which are reported to contain very high
  concentrations of anti-nutritional factors can
  cause adverse health effects.

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• There are reports from time to time of deaths
  after consumption of some type of beans despite
  cooking and also reports of renal and liver
  failure.
• There is a wide distribution of
  biologically-active constituents throughout the
  plant kingdom, particularly in plants used as
  animal feeding stuff and in human nutrition.
• Some of these plant chemicals have been shown to
  be deleterious to health or evidently
  advantageous to human and animal health if
  consumed in appropriate amounts after adequate
  processing.

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What are these ANFs ?

• Plants contain starch polysaccharides and
  non-starch polysaccharides (NSPs). Some of these
  polysaccharides are antinutritional factors.
• Starch is made up of glucose molecules connected
  together by a-glycosidic linkage and these are
  easily broken by endogenous enzymes in birds and
  mammals.
• All other glycosidic bonds are resistant to
  endogenous digestive enzymes of animals,
  however, they can be digested by microbe-derived
  enzymes.

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• NSPs contain sugars other than glucose and have
  linkages other than the a-linkages common in
  sugar. An example of an NSP is cellulose and they
  have ß-links and this difference in orientation
  makes them resistant to digestion by endogenous
  digestive enzymes of animals.
• Plants also contain anti- nutrients acquired from
  fertilizer and pesticides and several
  naturally-occurring chemicals.
• Some of these chemicals are known as secondary
  metabolites and they have been shown to be
  highly biologically active.

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• They include saponins, tannins, flavonoids,
  alkaloids, trypsin (protease) inhibitors,
  oxalates, phytates, haemagluttinins (lectins),
  cyanogenic glycosides, cardiac glycosides,
  coumarins and gossypol.
• These NSPs and Secondary metabolites adversely
  affect digestion in animals consuming them.
  Soluble NSPs affect the viscosity of the material
  in the digestive tract, which in turn, affects
  the ability of the digestive enzymes to reach
  their target.
• Absorption of any released nutrients is also
  reduced. This reduction in nutrient absorption
  results in reduced feed efficiency.

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Example for ANFs in Soyabeans -
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Classes of ANFs

• They are broadly classified into four groups -
• Factors affecting protein utilisation and
  digestion, such as protease inhibitors, tannins,
  lectins,
• Factors affecting mineral utilisation, which
  include phytates, gossypol pigments, oxalates,
  glucosinolates,
• Antivitamins,
• Miscellaneous substances such as mycotoxins,
  mimosine, cyanogens, nitrate, alkaloids,
  photosensitizing agents, phytoestrogens and
  saponins.

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• Also be classified according to their ability to
  withstand thermal processing-
• Heat labile factors include protease inhibitors,
  phytates, lectins, goitrogens and antivitamins.
• Heat stable factors are represented by saponins,
  non-starch polysaccharides, antigenic proteins,
  estrogens and some phenolic compounds.

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Applications in food industry

• Saponins - Used as flavourings in food and foam
  producing agents antioxidants for food use and
  in the production of spray dried powders of
  Vitamin E for the enrichment of foods and
  beverages.
• Flavonoids - Used in food processing industry to
  inhibit heat or chemical initiated lipid
  peroxidation as well as chelating metallic and
  superoxide ions.
• Flavones and leucoanthocyanidins - Impart very
  pleasant flavours in foods after processing and
  also impart a bitter taste in many soft drinks
  and bitter lemon brands.

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ANFs in fish feed

• Aquaculture has become the fastest growing food
  production sector of the world, with an average
  annual increase of about 10.
• To sustain such high rates of increase in
  aquaculture production, a matching increase in
  the levels of production of fish feed is
  required.
• Aqua feed production is currently one of the
  fastest expanding agricultural industries of the
  world, with annual growth rates in excess of 30
  per year.

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• Fish meal and fish-oil are mainly used as the
  feed for aquatic animals, but fish meal
  production requires raw materials either from
  capture fisheries or aquaculture production. So
  the aquafeed industries are turning towards
  alternative protein sources from plants for
  feeding the fishes.
• Most of the potential, alternative, plant-derived
  nutrient sources are known to contain a wide
  variety of antinutritional substances.

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• Important antinutrients present in some commonly
  used alternative fish feed ingredients -
• Soybean meal - Protease inhibitors, lectins,
  phytic acid, saponins, phytoestrogens,
  antivitamins, allergens
• Rapeseed meal - Protease inhibitors,
  glucosinolates, phytic acid, tannins
• Lupin seed meal - Protease inhibitors, saponins,
  phytoestrogens, alkaloids
• Pea seed meal - Protease inhibitors, lectins,
  tannins, cyanogens, phytic acid, saponins,
  antivitamins
• Sunflower oil cake - Protease inhibitors,
  saponins, arginase inhibitor
• Cottonseed meal- Phytic acid, phytoestrogens,
  gossypol, antivitamins, cyclopropenoic acid
• Mustard oil cake - Glucosinolates, tannins
• Sesame meal - Phytic acid, protease
  inhibitors

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Some common effects of alternative fish feed
ingredients on fish -

1. Soybean meal Fishes showed decreased growth
   performance and reduced amino acid absorption
   rate ( Dabrowski et al.,1989) and abnormal
   intestinal morphology in Atlantic salmon (van den
   Ingh., 1991).
2. Lupin seed meal Fishes showed lipid deposition
   in the liver and growth performance depression by
   50 ( Burel et al., 1998).
3. Rapeseed meal Rainbow trout fed with rapeseed
   meal showed significantly lower growth rate but
   higher protein efficiency ratio (Gomes et al.,
   1993).

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• Cottonseed meal Studies in Tilapia showed
  decrease in growth rate at all levels (Ofojekwu
  et al., 1984).
• Sunflower meal No significant difference in
  weight gain and growth rate (Sanz et al., 1994).
• Cassava or rice meal In carps, showed greater
  weight gain, better food utilisation and protein
  sparing (Ufodike and Matty, 1983).

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ANFs and immune system of fishes

• Energy and nutrients consumed with food are
  essential for maintaining optimal immune
  function. Without adequate nutrition, the immune
  system is deprived of the resources that are
  needed to defend the host against bacteria,
  viruses and parasites.
• Studies on the effects of ANFs over fish immune
  system are limited.
• ANFs present in alternative fish feed
  ingredients have shown some effects over the
  immune system of fishes

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• Soy protein concentrate diet have shown altered
  expression of immune genes in
• mid intestine (most genes upregulated, some
  downregulated),
• liver (most genes downregulated) and
• skeletal muscle (most genes downregulated),
  indicating both local and systemic immune
  responses to SPC (Tacchi et al., 2012).
• Combination of pea protein concentrate and
  soya-saponin induced gut inflammation and altered
  expression of immune genes i.e upregulation of
  cytokines, NFkB and TNF-alpha related genes and
  regulators of T-cell function, coupled with
  down-regulation of IFN-axis (Kortner et al.,
  2012).

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• Soybean meal diet induced gut inflammation at
  histological level and increased expression of
  immune-related genes, including GTPase IMAP
  family members, NF-kB-related genes and
  regulators of T cell and B cell function,
  indicating a rapid onset of disease (Sahlmann et
  al., 2013).
• Rapeseed meal have shown larger effects on immune
  gene expression with majority of genes related to
  processes of both innate and adaptive immunity
  upregulated, apart from T cell and leukotriene B4
  (LTB4) receptors that were downregulated (Morais
  et al., 2011).

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• Fishes fed with a high concentration of soyfeed
  diet have shown an immunological tolerance
  induction and also immunosuppression. Suppression
  of non-specific immune capacity was also
  apparent. (Burrells et al., 1999).
• Dietary manipulation by bioprocessed soymeal feed
  showed a significant decrease in the spontaneous
  macrophage killing activity and also decrease in
  respiratory burst activity (Kokou et al., 2012).

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• Use of alginate in Atlantic salmon have shown
  increased values of different non-specific immune
  mechanisms, which have been interpreted as
  inflammatory/hypersensitivity or
  immunostimulating effects (Gabrielsen and
  Austreng, 1998).
• Bean protein feed have shown induced gut
  inflammation at histological level and altered
  pathways associated with inflammatory and immune
  responses, suggesting ongoing disease (Krol et
  al. 2016).

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• In Atlantic salmon, soybean meal produced a
  decrease in mucosal enzymes, which were
  coincidental with an impaired feed conversion
  (Krogdahl et al., 2003).
• Partial replacement of fishmeal with rapeseed
  meal have shown no effect on serum lysozyme and
  total peroxide but fishes showed less tolerance
  to oxidative stress. (Dossou et al., 2018).

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• Atlantic salmon fed with glucosinolates showed
  decreased lice infection, increased expression of
  IFN related genes prior to infection and induced
  higher expression profiles of Type 1 immune genes
  late into the infection (Jodaa Holm et al.,
  2016).
• Increased number of IgM cells were found in
  fishes fed with saponins and phytosterols similar
  to other fishes exposed to antigenic proteins
  (Couto et al., 2015).

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References

• Emire, S.A., Jha, Y.K. and Mekam, F., 2013. Role
  of anti-nutritional factors in food industry.
  Beverage and Food World, 1, pp.23-28.
• Francis, G., Makkar, H.P. and Becker, K., 2001.
  Antinutritional factors present in plant-derived
  alternate fish feed ingredients and their effects
  in fish. Aquaculture, 199(3-4), pp.197-227.
• Gilani, G.S., Cockell, K.A. and Sepehr, E., 2005.
  Effects of antinutritional factors on protein
  digestibility and amino acid availability in
  foods. Journal of AOAC International, 88(3),
  pp.967-987.
• Soetan, K.O., 2008. Pharmacological and other
  beneficial effects of antinutritional factors in
  plants-A review. African journal of
  Biotechnology, 7(25).

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• Gilani, G.S., Xiao, C.W. and Cockell, K.A., 2012.
  Impact of antinutritional factors in food
  proteins on the digestibility of protein and the
  bioavailability of amino acids and on protein
  quality. British Journal of Nutrition, 108(S2),
  pp.S315-S332.
• Martin, S.A. and Król, E., 2017. Nutrigenomics
  and immune function in fish new insights from
  omics technologies. Developmental Comparative
  Immunology, 75, pp.86-98.
• Batista, S., Ozorio, R.O., Kollias, S.,
  Dhanasiri, A.K., Lokesh, J., Kiron, V., Valente,
  L.M. and Fernandes, J.M., 2016. Changes in
  intestinal microbiota, immune-and stress-related
  transcript levels in Senegalese sole (Solea
  senegalensis) fed plant ingredient diets
  intercropped with probiotics or immunostimulants.
  Aquaculture, 458, pp.149-157.
• Dossou, S., Koshio, S., Ishikawa, M., Yokoyama,
  S., Dawood, M.A., El Basuini, M.F., El-Hais, A.M.
  and Olivier, A., 2018. Effect of partial
  replacement of fish meal by fermented rapeseed
  meal on growth, immune response and oxidative
  condition of red sea bream juvenile, Pagrus
  major. Aquaculture.

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• Couto, A., Kortner, T.M., Penn, M., Bakke, A.M.,
  Krogdahl, Å. and Oliva-Teles, A., 2015. Dietary
  saponins and phytosterols do not affect growth,
  intestinal morphology and immune response of
  on-growing European sea bass (Dicentrarchus
  labrax). Aquaculture nutrition, 21(6),
  pp.970-982.
• Estruch, G., Collado, M.C., Peñaranda, D.S.,
  Vidal, A.T., Cerdá, M.J., Martínez, G.P. and
  Martinez-Llorens, S., 2015. Impact of fishmeal
  replacement in diets for gilthead sea bream
  (Sparus aurata) on the gastrointestinal
  microbiota determined by pyrosequencing the 16S
  rRNA gene. PLoS One, 10(8), p.e0136389.
• Kiron, V., Puangkaew, J., Ishizaka, K., Satoh, S.
  and Watanabe, T., 2004. Antioxidant status and
  nonspecific immune responses in rainbow trout
  (Oncorhynchus mykiss) fed two levels of vitamin E
  along with three lipid sources. Aquaculture,
  234(1-4), pp.361-379.
• Burrells, C., Williams, P.D., Southgate, P.J.,
  Crampton, V.O., 1999. Immunological,
  physiological and pathological responses of
  rainbow trout (Oncorhynchus mykiss) to increasing
  dietary concentrations of soybean proteins. Vet.
  Immunol. Immunopathol. 72, 277 288.

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Thank You