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Research Article
 

Effect of Dietary Supplementation of Chitosan on Blood Biochemical Profile of Laying Hens



Elvia Hernawan, Lovita Adriani, Andi Mushawwir, Chandrawati Cahyani and Darmawan
 
ABSTRACT

Background and Objective: Chitosan is a natural alkaline polysaccharide and widespread in nature. A study was conducted to evaluate the effect of dietary supplementation of chitosan on blood biochemical parameters like cholesterol, malondialdehyde (MDA), creatinine and total leucocytes. Methodology: One hundred laying phase hens, aged 28 weeks were used in the study. The birds were divided into 2 treatment groups each having 3 replicates of 10 birds each. Group I served as control and was fed basal diet. In Group II, basal diet was supplemented with chitosan at 150 ppm g–1 by spraying method. Results: The results showed that the chitosan inclusion in the diet of layer pullets significantly (p<0.05) lowered the total cholesterol (36.749±0.381 mg dL–1) when compared to the control group (43.030±0.352 mg dL–1). The dietary incorporation of chitosan significantly (p<0.05) decreased blood MDA levels (1.829±0.237 nmoles mL–1) of laying hens compared to control (2.553±0.379 nmoles mL–1). The creatinine levels also decreased significantly (p<0.05) in birds fed diet supplemented with chitosan, however, there was no effect on the total leucocyte count. Conclusion: The incorporation of chitosan in the diet had positive effect in terms of reducing the blood cholesterol and malondialdehyde levels of laying hens.

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  How to cite this article:

Elvia Hernawan, Lovita Adriani, Andi Mushawwir, Chandrawati Cahyani and Darmawan , 2017. Effect of Dietary Supplementation of Chitosan on Blood Biochemical Profile of Laying Hens. Pakistan Journal of Nutrition, 16: 696-699.

DOI: 10.3923/pjn.2017.696.699

URL: https://scialert.net/abstract/?doi=pjn.2017.696.699
 
Received: December 13, 2016; Accepted: June 21, 2017; Published: August 15, 2017


Copyright: © 2017. This is an open access article distributed under the terms of the creative commons attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are credited.

INTRODUCTION

Chitosan, a deacetylated chitin, is widespread in nature. The exoskeletons of arthropods such as crabs, shrimps, insects and other marine creatures in the Crustacean family are good sources of chitosan1-3. Chitosan is a natural alkaline polysaccharide with positive charge and also one of the most abundant natural polymers4-6. Unlike chitin, chitosan oligosaccharide (COS) is soluble in acidic solutions7 and it is partially digested in the gastrointestinal tract of monogastric animals8,9. Chitosan is commercially manufactured from chitin, in the process of deacetylation, by treating chitin with a strong solution of sodium hydroxide at an elevated temperature10. As chitosan and its oligosaccharide derivatives contain reactive, functional groups, that is, amino acids and hydroxyl groups, they have unlike chitin, antimicrobial11,12, anti-inflammatory13,14, anti-oxidative15,16, immunostimulatory17,18 and hypocholesterolemic19,20 properties.

Cholesterol level in meat has great importance as hypercholesterolemia is a risk factor for cardiovascular diseases such as atherosclerosis and myocardial infarctions, which is a common cause of mortality and morbidity in humans21,22. Many people limit meat consumption because of fear of elevated serum cholesterol concentrations23. Malondialdehyde (MDA) is the direct product of lipid peroxidation developed after radical attack and thus is an indicator of the extent of cell damage24. The MDA has been reported to cause fragmentation of destruction of cell membrane structure, deoxyribonucleic acid (DNA) and accelerate apoptosis25, thus warrants estimation of its levels as well. Creatinine is an important indicator of protein metabolism and its concentration is directly proportional to muscle mass, age, physical activity and diet26.

In view of the above, a study was conducted to evaluate the effect of dietary supplementation of chitosan on blood biochemical parameters like cholesterol, malondialdehyde (MDA), creatinine and total leucocytes.

MATERIALS AND METHODS

Birds and management: The experiment was carried out at CV Acum, Kuningan West Java, Indonesia. The study was conducted on 100 laying phase hens (having average body weight of 1452±23 g) aged 28 weeks. The birds were divided into 2 treatment groups each having 3 replicates of 10 birds each. Group I served as control and was fed basal diet. The composition of the basal diet was as follows: Total protein 18%, fat 8%, crude fiber 4%, methionine 0.4%, cysteine 0.3%, tryptophan 0.18%, lysine 0.9%, calcium 1% and phosphorus 0.5%. In Group II, basal diet was supplemented with chitosan at 150 ppm g–1 by spraying method.

Processing of chitosan: Low molecular weight chitosan with average molecular weight (Mw) of 7×103 Da (oligochitosan or irradiated chitosan) was provided by Center for Isotopes and Radiation Application (CIRA), National Nuclear Energy Agency (NNEA) Indonesia. The chitosan was prepared by irradiating the high molecular weight (Mw) chitosan of (1.5×105 Da) using gamma rays from Cobalt-60 source according to procedures in PATENT No IDP000034713 (15a).

Parameters recorded: Blood was collected after the end of trail (6 weeks) from all the birds. The parameters recorded were cholesterol, MDA, creatinine and total leucocytes. The cholesterol was estimated by the help of cholesterol kit using method of CHOD-PAP (Cholesterol Oxidase Phenylperoxidase Amino Phenozonphenol) as per Richmond27. The MDA was estimated using a modified test method thiobarbituric acid (TBA) by spectrophotometry as per Zainuri and Wanandi28. The collected blood sample 400 mL was treated with 200 mL of 20% trichloroacetic acid (TCA). The vortex was then applied to the resultant mixture and afterwards centrifuged at 5000 rpm for 10 min. The supernatant formed was collected and 400 mL of 0.67% TBA was added to it. The sample was again mixed thoroughly with vortex and incubated in water bath at a temperature of 96̊C for 10 min. The mixture was then removed and allowed to cool at room temperature. The absorbance was then read at a wavelength of 530 nm and MDA level was expressed as nmoles mL–1. The creatinine was estimated as per the method of Underwood29. Total leucocytes were determined by means of hematological analyzer (KT-6200 VET) and expressed as 103 mm–3.

Statistical analysis: Data collected were subjected to one way analysis of variance (ANOVA) as per Steel and Torrie30 and Duncan’s multiple range test31 was used to test the significance of difference between means considered significant at p<0.05.

RESULTS AND DISCUSSION

The results showed that the chitosan inclusion in the diet of layer pullets significantly (p<0.05) lowered the total cholesterol when compared to the control group. Several factors such as a diet rich in cholesterol has been reported to cause heart failure32.

Table 1:
Blood biochemical profile of laying hens fed chitosan in the diet
Means within the rows with different superscripts are significantly different (p>0.05)

Further, the high levels of cholesterol, particularly low-density lipoprotein (LDL), are mainly responsible for hypercholesterolemia22, which is a risk factor for cardiovascular diseases such as atherosclerosis and myocardial infection21. Therefore, inclusion of chitosan in the diet of poultry birds could be an important asset for health conscious people, however, the mechanism by which chitosan reduces blood cholesterol in poultry birds is not clear.

The dietary incorporation of cholesterol significantly (p<0.05) decreased blood MDA levels of laying hens. Malondialdehyde (MDA) is the direct product of lipid peroxidation developed after radical attack and thus is an indicator of the extent of cell damage24. The MDA has been reported to cause fragmentation of destruction of cell membrane structure, deoxyribonucleic acid (DNA) and accelerate apoptosis25. The results of present study thus indicated that chitosan helped the birds in alleviating day to day stress as indicated by the decreased MDA levels in chitosan fed group.

The creatinine levels also decreased significantly (p<0.05) in birds fed diet supplemented with chitosan. High creatinine levels are indicative of increased protein metabolism or kidney problems. Therefore, it is suggested that the chitosan incorporation in the diet of laying hens has no adverse effect on the kidneys of birds. Moreover, there was no effect on the total leucocyte count as a result of dietary incorporation of chitosan in layer birds, indicating that there was not any pathological condition associated with the inclusion of chitosan in the diet (Table 1).

CONCLUSION

The incorporation of chitosan in the diet of laying hens had positive effect in terms of reducing the blood cholesterol and malondialdehyde levels without having any pathological effect on the birds. However, further studies are needed to evaluate the mechanism by which chitosan helps in achieving such positive effects.

ACKNOWLEDGMENT

The authors would like to express special gratitude to the students in chitosan group as well the principal who helped a lot during the present study.

REFERENCES
Abeywardena, M.Y., 2003. Dietary fats, carbohydrates and vascular disease: Sri Lankan perspectives. Atherosclerosis, 171: 157-161.
CrossRef  |  Direct Link  |  

Crini, G., 2005. Recent developments in polysaccharide-based materials used as adsorbents in wastewater treatment. Prog. Polym. Sci., 30: 38-70.
CrossRef  |  Direct Link  |  

Duncan, D.B., 1955. Multiple range and multiple F tests. Biometrics, 11: 1-42.
CrossRef  |  Direct Link  |  

Hirano, S., C. Itakura, H. Seino, Y. Akiyama, I. Nonata, N. Kanbara and T. Kawahami, 1990. Chitosan as an ingredient for domestic animal feeds. J. Agric. Food Chem., 38: 1214-1217.
CrossRef  |  Direct Link  |  

Holappa, J., M. Hjalmarsdottir, M. Masson, O. Runarsson and T. Asplund et al., 2006. Antimicrobial activity of chitosan N-betainates. Carbohydr. Polym., 65: 114-118.
CrossRef  |  Direct Link  |  

Huang, R.L., Y.L. Yin, G.Y. Wu, Y.G. Zhang and T.J. Li et al., 2005. Effect of dietary oligochitosan supplementation on ileal digestibility of nutrients and performance in broilers. Poult. Sci., 84: 1383-1388.
CrossRef  |  Direct Link  |  

Kim, K.W. and R.L. Thomas, 2007. Antioxidative activity of chitosans with varying molecular weights. Food Chem., 101: 308-313.
CrossRef  |  Direct Link  |  

Knaul, J.Z., S.M. Hudson and K.A.M. Creber, 1999. Crosslinking of chitosan fibers with dialdehydes: Proposal of a new reaction mechanism. J. Polym. Sci. Part B: Polym. Phys., 37: 1079-1094.
CrossRef  |  Direct Link  |  

Krieger, M., 1998. The best of cholesterols, the worst of cholesterols: A tale of two receptors. Proc. Natl. Acad. Sci. USA., 95: 4077-4080.
PubMed  |  Direct Link  |  

Li, H.Y., S.M. Yan, B.L. Shi and X.Y. Guo, 2009. Effect of chitosan on nitric oxide content and inducible nitric oxide synthase activity in serum and expression of inducible nitric oxide synthase mRNA in small intestine of broiler chickens. Asian-Aust. J. Anim. Sci., 22: 1048-1053.
CrossRef  |  Direct Link  |  

Liu, J., J. Zhang and W.S. Xia, 2008. Hypocholesterolaemic effects of different chitosan samples in vitro and in vivo. Food Chem., 107: 419-425.
CrossRef  |  Direct Link  |  

Ma, P., H.T. Liu, P. Wei, Q.S. Xu, X.F. Bai, Y.G. Du and C. Yu, 2011. Chitosan oligosaccharides inhibit LPS-induced over-expression of IL-6 and TNF-α in RAW264.7 macrophage cells through blockade of Mitogen-Activated Protein Kinase (MAPK) and PI3K/Akt signaling pathways. Carbohydr. Polym., 84: 1391-1398.
CrossRef  |  Direct Link  |  

Nishimura, K., S.I. Nishimura, N. Nishi, F. Numata, Y. Tone, S. Tokura and I. Azuma, 1985. Adjuvant activity of chitin derivatives in mice and guinea-pigs. Vaccine, 3: 379-384.
CrossRef  |  Direct Link  |  

Okamoto, Y., M. Nose, K. Miyatake, J. Sekine, R. Oura, Y. Shigemasa and S. Minami, 2001. Physical changes of chitin and chitosan in canine gastrointestinal tract. Carbohydr. Polym., 44: 211-215.
CrossRef  |  Direct Link  |  

Ormrod, D.J., C.C. Holmes and T.E. Miller, 1998. Dietary chitosan inhibits hypercholesterolaemia and atherogenesis in the apolipoprotein E-deficient mouse model of atherosclerosis. Atherosclerosis, 138: 329-334.
CrossRef  |  Direct Link  |  

Puvaca, N., L. Kostadinovic, S. Popovic, J. Levic and D. Ljubojevic et al., 2016. Proximate composition, cholesterol concentration and lipid oxidation of meat from chickens fed dietary spice addition (Allium sativum, Piper nigrum, Capsicum annuum). Anim. Prod. Sci., 56: 1920-1927.
CrossRef  |  Direct Link  |  

Richmond, W., 1973. Preparation and properties of a cholesterol oxidase from Nocardia sp. and its application to the enzymatic assay of total cholesterol in serum. Clin. Chem., 19: 1350-1356.
PubMed  |  Direct Link  |  

Schaefer, E.J., A.H. Lichtenstein, S. Lamon-Fava, J.R. McNamara and J.M. Ordovas, 1995. Lipoproteins, nutrition, aging and atherosclerosis. Am. J. Clin. Nutr., 61: 726S-740S.
PubMed  |  Direct Link  |  

Shahidi, F., J.K.V. Arachchi and Y.J. Jeon, 1999. Food applications of chitin and chitosans-A review. Trends Food Sci. Technol., 10: 37-51.
CrossRef  |  Direct Link  |  

Singla, A.K. and M. Chawla, 2001. Chitosan: Some pharmaceutical and biological aspects-an update. J. Pharm. Pharmacol., 53: 1047-1067.
CrossRef  |  PubMed  |  Direct Link  |  

Steel, R.G.D. and J.H. Torrie, 1980. Principles and Procedures of Statistics: A Biometral Approach. McGraw-Hill Book Company, New York.

Szabo, A., M. Mezes, P. Horn, Z. Suto, G.Y. Bazar and R. Romvari, 2005. Developmental dynamics of some blood biochemical parameters in the growing turkey (Meleagris gallopavo). Acta Vet. Hung., 53: 397-409.
CrossRef  |  PubMed  |  Direct Link  |  

Underwood, J.C.E., 1997. General and Systematic Pathology. Publishers Books Medicine EGC., Jakarta, Indonesia.

Wald, N.J. and M.R. Law, 1995. Serum cholesterol and ischaemic heart disease. Atherosclerosis, 118: S1-S5.
CrossRef  |  PubMed  |  Direct Link  |  

Wang, H.F., X.H. Zhong, W.Y. Shi and B. Guo, 2011. Study of malondialdehyde (MDA) content, superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in chickens infected with avian infectious bronchitis virus. Afr. J. Biotechnol., 10: 9213-9217.
Direct Link  |  

Wang, S.Q. and C.S. Zhang, 2004. Chitin, chitosan and their application in aquaculture. Feed Res., 5: 25-28.
Direct Link  |  

Xu, C.L. and Y.Z. Wang, 2005. Application of chitin in aquiculture. China Feed, 7: 30-32.
Direct Link  |  

Yen, M.T., J.H. Yang and J.L. Mau, 2008. Antioxidant properties of chitosan from crab shells. Carbohydr. Polym., 74: 840-844.
CrossRef  |  Direct Link  |  

Yoon, H.J., M.E. Moon, H.S. Park, S.Y. Im and Y.H. Kim, 2007. Chitosan Oligosaccharide (COS) inhibits LPS-induced inflammatory effects in RAW 264.7 macrophage cells. Biochem. Biophys. Res. Commun., 358: 954-959.
CrossRef  |  Direct Link  |  

Zaharoff, D.A., C.J. Rogers, K.W. Hance, J. Schlom and J.W. Greiner, 2007. Chitosan solution enhances both humoral and cell-mediated immune responses to subcutaneous vaccination. Vaccine, 25: 2085-2094.
CrossRef  |  Direct Link  |  

Zainuri, M. and S.I. Wanandi, 2012. [Spesific activity of Manganese Superoxide Dismutase (MnSOD) and catalase in the rat liver induced systemic hypoxia: Relationship with oxidative damage]. Media Litbang Kesehatan, 22: 87-92, (In Indonesian).
Direct Link  |  

Zheng, L.Y. and J.F. Zhu, 2003. Study on antimicrobial activity of chitosan with different molecular weights. Carbohydr. Polym., 54: 527-530.
CrossRef  |  Direct Link  |  

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