Majid Toghyani +, Sayedhamid Zarkesh,, Mahmoud Shivazad - and Abbasali Gheisari.

Transcription

1 The Journal of Poultry Science : Majid Toghyani + Sayedhamid Zarkesh Mahmoud Shivazad - and Abbasali Gheisari + Department of Animal Science Khorasgan Azad University Esfahan Iran Dpartment of Biology Faculty of Science Esfahan University Iran - Department of Animal Science College of Agriculture University of Tehran Karaj Iran Department of Animal Science Esfahan Agricultural Research Center Esfahan Iran The present study was conducted with two hundreds and forty male broiler chicks to evaluate the e#ect of chromium (Cr) from Cr picolinate on immune responses of heat-stressed broiler chicks In a completely randomized design diets were supplemented with (control) / and ppb Cr Antibody titers against Newcastle and Influenza virus at +2 and -d heterophil to lymphocyte ratios albumin to globulin ratios and concentration of immunoglobulin G in serum were measured at + and d On day thymus spleen and bursa of Fabricius were collected weighed and expressed as a percentage of live body weight Antibody titers against Newcastle and Influenza virus were elevated at - d of age in broiler chicks fed supplemental Cr Heterophil to lymphocyte ratios decreased in broiler chicks fed and ppb supplemental Cr Concentration of immunoglobulin G in serum was increased by Cr supplementation especially at level of ppb Albumin to globulin ratios and weights of lymphoid organs were not a#ected by supplemental Cr The results suggest that Cr supplementation improves some immune responses of heat-stressed broiler chicks Key words: broilers chromium heat stress immune response J Poult Sci : Introduction Chromium (Cr) is an essential element required for carbohydrate fat and protein metabolism (Anderson ) Trivalent Cr is a component of glucose tolerance factor (GTF) which participate in glucose metabolism by enhancing the e#ect of insulin (Anderson +321; Mertz +33-) Stress and disease increased urinary excretion of Cr (Borel et al +32; Anderson et al +322) and may exacerbate a marginal Cr deficiency Immunological function has been enhanced by trivalent Cr and its e#ects seem more pronounced during times of stress (Borgs and Mallard +332) Alleviating stressassociated immunosuppression thus may be one of the mechanisms by which Cr acts (Burton et al +33-) Improvements in immune response have been observed when organic forms of Cr were supplemented to broilers (Luo et al +333 ) stressed feeder calves (Chang and Mowat +33; Moonsie- Shageer and Mowat +33-) and dairy cows (Burton et al +33-) No recommendations for Cr are currently listed for poultry (NRC ) and most poultry diets are basically composed of plant origin ingredients which have usually low content of Cr (Giri et al +33) This study was conducted to investigate the e#ects of di#erent levels of Cr picolinate on immune responses in heat-stressed broiler chicks Materials and Methods Two hundred and forty male broiler chicks (Ross Received: January 3 1 Accepted: April 3 1 Correspondence: M Toghyani Department of Animal Science Khorasgan Azad University Esfahan Iran ( toghiani@hotmailcom)

2 Toghyani et al: Chromium and Immune Response 331-2) were allocated to four treatments in a completely randomized design Dietary treatments supplemented with (control) / and ppb Cr in the form of Cr picolinate (contain +1 Cr) Each dietary treatment was randomly allocated to four replicates of +/ chicks each Chicks were raised from + to days of age in controlled house with mean value of daily temperature -- - The chicks were fed a maize-soybean meal starter diet (+3 g protein -/ kcal ME kg + )upto + days and a grower diet ( g protein - kcal ME kg + )upto days The basal diets in mash form were formulated to meet or exceed the nutrient requirements of broiler chickens (NRC +33) Cr contents were -/ and -30 ppm in starting and finishing basal diets respectively as measured by atomic absorption spectrometer with a graphite furnace (Perkin-Elmer AAnalyst 0 USA)Thediets and fresh water were provided ad libitum Chicks were maintained on a h constant light schedule All chicks were intramuscularly immunized with killed vaccine of Newcastle and Avian Influenza (H 3 N) viruses at age of 2 days Live Newcastle disease vaccine (Nobilis ND Lasota) was administered orally (drinking water) at d On days +2 and - blood samples were collected from the wing vein of three birds per replicate and serum antibody titers against Newcastle and Influenza viruses were determined by haemagglutination inhibition (HI) test and were expressed as the logarithm base On days + and blood samples of three chicks from each replicate were obtained from the wing vein using heparin as anticoagulant Blood smears were prepared using May-Greenwald-Giemsa stain and heterophil to lymphocyte ratios were based on a total of + cells (Gross and Siegel +32-) Serum protein fractions were separated by electrophoresis (+/V// min) in cellulose polyacetate Five fractions were obtained (albumin a + a b and g globulin) and albumin to globulin ratio was calculated Quantitative ELISA immunoassay kits for measurement of chicken immunoglobulin (IgG) from Bethyl Laboratories (Montgomery TX USA) were used to measure the IgG in serum samples of twelve chicks per treatment The result was monitored as OD at / nm The IgG concentrations (ng/ml serum) were calculated from a standard curve as suggested by the company Three chicks from each replicate were slaughtered on day and lymphoid organs such as thymus spleen and bursa of Fabricius were collected weighed and expressed as a percentage of live body weight Data were analyzed by analysis of variance procedures appropriate for a completely randomized design using the GLM procedures of SAS (+331) Significant di#erences (P /) among treatment means were determined using multiple range test (Duncan +3//) Results The e#ects of supplemental Cr on antibody titers against Newcastle and Influenza virus are shown in Table + Antibody titers against Newcastle and Influenza virus at - days of age in broiler fed supplemental Cr were higher than in broiler chicks fed control diets (P /) But at +2 days of age there was no significant di#erence in the antibody titer among the treatments Heterophil to lymphocyte ratios were significantly lower (P /) at+ din broiler chicks fed and ppb Cr supplementation (Table ) No significant di#erences in albumin to globulin ratio were observed at + and d Dietary supplemental Cr tended to increase concentration of IgG in serum at + and d and supplement of ppb Cr resulted in significant (P /) increase in IgG concentration at d (Table ) The weights of lymphoid organs were not a#ected in broiler chicks fed diets with di#erent levels of supplemental Cr (Table -) Discussion Dietary Cr supplementation tended to increase antibody titers against Newcastle and Influenza virus and supplement of and ppb Cr resulted in significant increase of antibody titer at - d Elevated antibody titer against Newcastle disease was reported in broiler chicks under heat stress with supplement of or + ppm Cr either in the form of CrCl - or yeast (Guo et al +333) Lee et al (-) reported antibody titer against Infectious bronchitis was improved in broiler chicks fed ppb Cr picolinate Improved immune responses against virulent antigen were also reported in weanling pig (Lee et al +331) or calf (Burton et al +33) with dietary Cr supplement Zulkifli et al () reported antibody production in young broiler chicks

3 332 J Poult Sci (-) Table + E#ect of chromium supplementation on antibody titer against Newcastle and Influenza virus at di#erent ages / Newcastle (log HI titer) Influenza (log HI titer) +2 d - d +2 d - d /3 4++ b 4-- ab 42 a 41+ a c b 40+ a 4/ ab 4+30 Means in the same column with no common superscripts di#er significantly (P /) Table E#ect of chromium supplementation on heterophil to lymphocyte ratio (H/L) albumin to globulin ratio (A/G) and immunoglobulin G (IgG) at di#erent ages / H/L A/G IgG (ng/ml) + d d + d d + d d 40+ a 4/0 ab 4/+ b 4/ b /0/ 4/ Means in the same column with no common superscripts di#er significantly (P /) / /4 b /40 b 140 ab -4 a +4 Table - E#ect of chromium supplementation on lymphoid organ weight (percentage of live body weight) at days of age / Bursa of Fabricius Spleen Thymus 4+// / 42 4/3+ 40+/ 40 4/2-4/ Means in the same column with no common superscripts di#er significantly (P /) were decreased in heat stress condition This reduction could be indirectly due to an increase in inflammatory cytokines under stress which stimulates the hypothalamic production of corticotrophin releasing factor (Ogle et al +331) Corticotrophin releasing factor is known to increase adrenocorticotropic hormone from the pituitary; adrenocorticotropic hormone then stimulates corticosterone production from the adrenal gland Corticosterone inhibits antibody production (Gross +33) A reduction in serum cortisol level of heat-stressed broilers fed Cr picolinate was reported by Sahin et al () In this study the increases in lymphocytes count and consequently decrease in heterophil to lymphocyte ratios in broiler chicks fed and ppb Cr were observed at + d (Table ) The heterophil to lymphocyte ratio has been accepted as a reliable index for determining stress in poultry (Gross and Siegel +32-; McFarlane and Curtis +323) It has also been found that exposure of birds to heat stress results in an increase in the heterophil to lymphocyte ratio (Altan et al -; Zulkifli et al -) Gross and Siegel (+32-) found that the number of heterophils increased in the blood of corticosteronefed chicks The increases in lymphocyte counts and decreases in heterophil to lymphocyte ratios by Cr supplementation in heat stressed chicks in the present study may be attributed to decreased glucocorticoid secretion Serum IgG level in broiler chicks fed Cr supplementation were higher than chicks fed control

4 Toghyani et al: Chromium and Immune Response 333 diet These results are in agreement with most experiments involving supplemental Cr Uyanik et al () observed IgG and IgM titers tended to be higher in broiler chicks fed Cr supplementation in the form of CrCl - Total IgG was increased in stressed feeder calves receiving supplemental Cr nicotinate (Kegley and Spears +33/) Total IgG+ and IgM were increased after transport stress in calves supplemented with high-cr yeast (Chang and Mowat +33; Moonsie-Shageer and Mowat +33-) However these results contradict with findings of Kegley et al (+331) They reported total IgG was decreased with supplementation of Cr nicotinate in stressed calves The exact mechanism by which Cr enhances the immune system is not known However one of the consistent results of the studies was that Cr reduced serum cortisol levels Cortisol the most important glucocorticoid has been found to be immunosuppressive inhibiting the production and actions of antibodies lymphocyte function and leucocyte population (Roth and Kaeberle +32; Munck et al +32) In conclusion supplemental dietary Cr particularly at level of ppb improved some immune responses and can be used as additives in heatstressed broiler chicks diet but it still needs more investigation References Altan O Pabuccuoglu A Altan A Konyalioglu S and Bayraktar H E#ect of heat stress on oxidative stress lipid peroxidation and some stress parameters in broilers British Poultry Science : // // - Anderson RA Chromium In: Trace Elements in Human and Animal Nutrition pp / Academic Press New York +321 Anderson RA Chromium as an essential nutrient for humans Regular Toxicology and Pharmacology 0: S-/ S Anderson RA Bryden NA Polansky MM and Deuster PA Exercise e#ects on chromium excretion of trained and untrained men consuming a constant diet Journal of Applied Physiology 0: 3 / +322 Borel JS Majerus TC Polansky MM Mozer PB and Anderson RA Chromium intake and urinary Cr excretion of trauma patients Biological Trace Element Research 0: Borgs P and Mallard BA Immune-endocrine interactions in agricultural species: Chromium and its e#ect on health and performance Domestic Animal Endocrinology +/: Burton JL Mallard BA and Mowat DN E#ects of supplemental chromium on immune responses of periparturient and early lactation dairy cows Journal of Animal Science 1+: +/- +/ Burton JL Mallard BA and Mowat DN E#ects of supplemental chromium on antibody responses of newly weaned feedlot calves to immunization with infectious bovine rhinotracheitis and parainfluenza - virus Canadian Journal of Veterinary Research /2: +2 +/+ +33 Chang X and Mowat DN Supplemental chromium for stressed and growing feeder calves Journal of Animal Science 1: //3 +33 Duncan DB Multiple range and multiple F test Biometrics ++: + +3// Giri J Usha K and Sunita T Evaluation of the selenium and chromium content of plants foods Plant Foods Human Nutrition : 3 /3 +33 Gross WB E#ect of short-term exposure of chickens to corticosterone on resistance to challenge exposure with Escherichia coli and antibody response to sheep erythrocytes American Journal of Veterinary Research /-: Gross WB and Sigel PS Evaluation of heterophil to lymphocyte ratio as a measure of stress in chickens Avian Disease 1: Guo YL Luo XG Hao ZL Liu B Chen JL Gao FS and Yu SX E#ect of chromium on growth performance serum biochemical traits immune function and carcass quality of broiler chickens Scientia Agriculture Sinica -: Kegley EB and Spears JW Immune response glucose metabolism and performance of stressed feeder calves fed inorganic or organic chromium Journal of Animal Science 1-: / Kegley EB Spears JW and Brown TT E#ect of shipping and chromium supplementation on performance immune response and disease resistance of steers Journal of Animal Science 1/: +3/ Lee DN Yen HT Shen TF and Chen BJ The e#ects of chromium picolinate supplementation on growth performance and immunity response of weanling pigs Journal of Chinese Society of Animal Science 0: Lee DN Wu FY Cheng YH Lin RS and Wu PC E#ect of dietary chromium picolinate supplementation on growth performance and immune responses of broilers Asian- Australian Journal of Animal Science +0: Luo X Guo YL Liu B Hao ZL Chen JL Gao FS and Yu SX E#ect of dietary chromium on growth serum biochemical traits and immune responses of broiler chicks during - weeks of age Acta Veterinary et Zootechnica Sinica -: McFarlane JM and Curtis SE Multiple concurrent stressors in chicks - E#ects on plasma corticosterone and the heterophil to lymphocyte ratio Poultry Science 02: / / Mertz W Chromium in human nutrition: A review Journal of Nutrition +-: Moonsie-Shageer S and Mowat DN E#ect of level of supple-

5 334 J Poult Sci (-) mental chromium on performance serum constituents and immuno status of stressed feeder calves Journal of Animal Science 1+: Munck A Guyre PM and Holbrook NJ Physiological functions of glucocorticoids in stress and their relation to pharmacological actions Endocrinology Review /: / +32 National Research Council Nutrient requirements of poultry 3 th rev ed National Academy Press Washington DC +33 National Research Council The role of chromium in animal nutrition National Academy Press Washington DC +331 Ogle CK Valente JF Guo X Li BG Ogle JD and Alexander JW Thermal injury induces the development of inflammatory macrophages from nonadherent bone marrow cells Inflammation +: /03 / Roth JA and Kaeberle ML The e#ect of glucocorticoids on the bovine immune system Journal of American Veterinary Medicine Association +2: - -/ +32 Sahin K Sahin N Onderci M Gursu F and Cikim G Optimal dietary concentration of chromium for alleviating the e#ect of heat stress on growth carcass qualities and some serum metabolites of broiler chickens Biological Trace Element Research 23: /- 0 SAS Institute SAS Users Guide Version 00 review edition Cary NC +331 Uyanik F Atasever A Ozdmar S and Aydin F E#ects of dietary chromium chloride supplementation on performance some serum parameters and immune response in broilers Biological Trace Element Research 3: 33 ++/ Zulkifli I Che Norma MT Israf DA and Omar AR The e#ect of early age feed restriction on subsequent response to high environmental temperatures in female broiler chickens Poultry Science 13: Zulkifli I Liewa PK Israf DA Omar AR and Hair-Bejo M E#ects of early age feed restriction and heat conditioning on heterophil to lymphocyte ratios heat shock protein 1 expression and body temperature of heat-stressed broiler chickens Journal of Thermal Biology 2: +1 -