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1 VETERINARY RESEARCH INTERNATIONAL Journal homepage: ORIGINAL ARTICLE Effect of herbal vitamin E-selenium with organic chromium on growth, haematological and parasitological parameters of broiler chicken during mixed Eimeria infection V.S. Singh 1*, Jyoti Palod 2, Stuti Vatsya 3 and S.K. Shukla 4 Department of Veterinary Parasitology, College of Veterinary and Animal Sciences, G.B. Pant University of Agriculture and Technology, Pantnagar (Uttarakhand). INDIA. *Corresponding Author: V. S. Singh singhvs65@gmail.com ( 1 Assistant Professor, 2 Associate Professor, Department of Livestock Management and Production; 3 Associate Professor, 4 Professor, Department of Veterinary Medicine) Received: 10/05/2013 Revised: 11/08/2013 Accepted: 17/08/2013 Abstract A feeding trial of 5-weeks duration was conducted on day old seventy two broiler chicks to study the effect of herbal vitamin E-selenium with organic chromium on growth performance, haematological and parasitological parameters. At 21 days of age, the birds were randomly divided into three groups- A (uninfected control), B (infected untreated control) and C (infected and herbal vitamin E-selenium with organic chromium treated) (group B and C birds were experimentally infected with sporulated oocysts of mixed Eimeria species). During pre-infection period (0-21 days) weight gain and feed intake were significantly higher in herbal vitamin E-selenium with organic chromium supplemented group as compared to control groups while reverse trend was seen for feed conversion ratio (FCR). No significant difference in feed intake of broilers of different groups was observed during days of age (8-14 days post-infection). However, weight gain was maximum in supplemented group and minimum in infected control group. FCR showed just the reverse trend to that of weight gain. On 6 th day post-infection, haemoglobin and packed cell volume differed significantly among the broilers of 3 groups being maximum in uninfected control followed by vitamin E-selenium with organic chromium supplemented group and minimum in infected control group. Parasitological parameters in terms of oocyst index, lesion score, per cent protection against lesion, per cent faecal score, per cent weight gain, performance index, average oocyst production and per cent reduction in oocyst production revealed a significant improvement in all these parameters in vitamin E-selenium with organic chromium supplemented groups as compared to infected control group. It can be suggested that the combination of herbal vitamin E-selenium with organic chromium has the potential to provide protection in broiler chickens against coccidiosis induced alteration in growth, haematological and parasitological parameters. Key Words: Broiler growth, herbal vitamin E-selenium, organic chromium, Eimeria infection, haematological and parasitological parameters. Introduction Among the various nutrients required by the poultry, vitamin E and selenium play a vital role in the development and maintenance of the defense system (Marsh et al., 1982). Vitamin E and selenium are integral part of body s natural antioxidant system and have synergistic action. They protect the body cells from oxidative dama- ge, which is an ongoing phenomenon especially under disease and other stress conditions. Dietary supplementation with selenium or vitamin E has been reported to reduce mortality and increase body weight gain of non-immunized chicken infected with Eimeria tenella (Colnago et al., 1984).Chromium in the form of chromium picol-

2 inate also decreases mortality (Hossain et al., 1998) and increases immune response in broilers (Lee et al., 2003). Supplementary dietary chromium is recommended by NRC (1997) for animals undergoing environmental stress. Chromium has also possesses antioxidant properties in vivo (Tezuka et al., 1991). Most of the research work so far has been carried out on effect of vitamin E and selenium on immune response (Kidd, 2004), livability (Colnago et al., 1984) and growth performance of broilers during coccidial infection. Similarly, research work on use of organic chromium in poultry has been conducted as an anti stress agents (Sahin et al., 2001) as well as on its antioxidant role. Reports regarding research work on supplementation of herbal vitamin E-selenium along with organic chromium for the control of coccidiosis were difficult to fish out. Therefore, aim of the present experiment was to evaluate the effect of herbal Vitamin E-Selenium along with organic chromium on growth performance, haematological and parasitological parameters of broiler chicks experimentally infected with mixed sporulated oocysts of Eimeria species. Materials and Methods A total of seventy two day-old Vencobb broiler chicks were randomly allotted into 3 groups - A, B and C each with 2 replicates of 12 chicks each and were maintained in battery brooders from day old upto 35 days of age. Chicks were vaccinated against Newcastle disease on 7 th day of age. Standard broiler diet without anticoccidial was provided to the broilers of groups A and B while broilers of group C were provided 1 g herbal vitamin E-selenium/kg of feed along with 500 ppb organic chromium/litre of water. Feed and water were provided ad lib throughout experimental period. Mixed isolate of oocyst cultures were collected from litter, droppings, intestinal contents and mucus membranes of the infected birds during the outbreak of coccidiosis at poultry farms of Kashipur, Uttarakhand (India). The oocysts thus obtained were kept in 2.5% potassium dichromate solution for sporulation. The number of oocysts was counted in McMaster chamber (Davies et al., 1963). On day 21, broilers of group A were provided 1 ml plain water while those of groups B and C were provided 1 ml suspension containing 1,50,000 sporulated oocysts of mixed Eimeria species i.e. E. tenella (49%), E. acervulina (16%), E. necatrix (15%), E. maxima (11%) and E. mitis (9%). Body weights of broilers were recorded at the start of experiment. Weekly body weight, feed intake and daily mortality were recorded for each replicate group of treatments upto the age of 35 days. On days 6 and 14 post-infection, blood in heparinized vials was collected from 3 chicks randomly selected from each replicate for determination of haemoglobin (Hb) and packed cell volume (PCV) (Schalm, 1965). To evaluate the anticoccidial efficacy of feed supplements, percent survival was calculated for different groups. For parasitological parameters faecal score (%) was calculated from 4 th to 10 th day post-infection, per cent weight gain and performance index were calculated as per the method of Morehouse and Baron (1970). Faecal oocyst count was carried out from 4 th to 14 th day post-infection by collecting faeces from each infected sub-group (Davies et al., 1963) and hence, average oocyst count and reduction in oocyst production were calculated. Four broilers of each infected sub-group were sacrificed on day 6th post-infection to study the lesion score (Johnson and Reid, 1970) and oocyst index (Hilbrich, 1978) in the intestine of each bird. Statistical analysis for growth and haematological parameters was carried out using one way ANOVA technique as described by Snedecor and Cochran (1994). For parasitological studies, Cochran s approx t test was used. Results and Discussion Growth performance: Growth performance of broilers in terms of body weight gain, feed intake and feed conversion ratio (FCR) are presented in Table 1. During pre-infection period (0-21 days) weight gain and feed intake were significantly higher in herbal vitamin E-selenium with organic chromium supplemented group as compared to control groups while reverse trend was seen for FCR. It indicates positive influence of herbal vitamin E-selenium with organic chromium 26

3 supplementation on growth of broilers. Similar observations on effect of vitamin E-selenium on growth of broilers were recorded by Lohakare et al. (2004). While, Lee et al. (2003) found that chromium picolinate supplementation improved feed efficiency in broilers. It was observed that during days of age (0-7 days post-infection), weight gain and feed intake were maximum and significantly higher in control non-infected group, while, minimum and significantly lower in infected control group. Broilers supplemented with herbal vitamin E-selenium with organic chromium showed better growth than control infected broilers. Colnago et al. (1984) also found that dietary selenium and vitamin E supplementation increased body weight gain in chickens infected with E. tenella. No significant difference in feed intake of broilers of different groups was observed during days of age (8-14 days post-infection). However, weight gain was maximum in supplemented group and minimum in infected control group. FCR showed just the reverse trend to that of weight gain. Overall performance for post-infection period (0-14 days post-infection) revealed better weight gain in supplemented group as compared to control groups. Feed intake was minimum in infected control group while other 2 groups had non-significant difference in feed intake; FCR was significantly higher in infected control group than other 2 groups. The results of this experiment confirm the reports of various workers (Vegad, 2004) that infection with coccidial protozoan significantly reduces the performance of broilers. Haematological parameters: Perusal of results on haemoglobin concentration and packed cell volume (Table 1) obviously points that on 6 th day post-infection, both these parameters differed significantly among the broilers of all 3 groups being maximum in uninfected control followed by vitamin E-selenium with organic chromium supplemented group and minimum in infected control group. Reduced haemoglobin concentration and PCV values at 6 th day post infection in infected group were due to the intestinal haemorrhage resulting from the liberation of second generation merozoites which caused sloughing of intestinal mucosa with discharge of large amount of blood (Nayak and Rai, 1985). The results indicate that broiler chicken supplemented diet exhibited less blood loss despite coccidial infection. This might be due to the supplemented vitamin E-selenium increase in oxygen carrying capacity of haemoglobin and organic chromium act as antioxidant. Colnago et al. (1984) and Singh et al. (2008) also observed that vitamin E supplementation of chickens infected with coccidia resulted in significantly higher PCV. On 14 th day post-infection non-significant difference in haemoglobin concentration and PCV values were noted among broilers of different groups. Haemoglobin and PCV values were restored to normal due to the termination of haemorrhagic phase of coccidiosis in both infected control and supplemented infected groups (Natt and Herrick, 1955). Elevated haemoglobin levels in broilers supplemented with organic selenium have also been reported by Choct et al. (2004). Parasitological parameters: Parasitological parameters in terms of oocyst index, lesion score, per cent protection against lesion, per cent faecal score, per cent weight gain, performance index, average oocyst production and per cent reduction in oocyst production clearly revealed (Table 2) that there was significant improvement in all these parameters in vitamin E-selenium with organic chromium supplemented groups as compared to infected control group. The present study confirms the reports of earlier researchers (Colnago et al., 1984) who used vitamin E-selenium in diet of chickens infected with Eimeria tenella. Contrary to this, Allen et al. (1998) found that tocopherol at 8 ppm level significantly reduced lesion scores and oocyst output from E. maxima infection while it had little effect on E. tenella infection. The favourable effect of vitamin E and selenium could be attributed to their protective role as antioxidant. The studies conducted so far have shown that better results are obtained on administration of vitamin E and selenium in combinations rather than alone (Colnago et al., 1984). 27

4 Table 1: Effect of herbal vitamin E-selenium with organic chromium on growth performance and haematological parameters of broilers chicken infected with mixed Eimeria spp. Parameter Group I Group II Group III S.E. Weight gain 0-21 days (pre-infection period) b b a days (0-7 days post infection) a c b days (8-14 days post infection) b c a days (0-14 days post infection) b c a days (overall mean) b c a 8.03 Feed Intake 0-21 days (Pre-infection period) b b a days (0-7 days post-infection) a c b days (8-14 days post-infection) b c a days (0-14 days post-infection) a b a days (Overall mean) b c a Feed Conversion Ratio (FCR) 0-21 days (Pre-infection period) 1.62 a 1.61 a 1.54 b days (0-7 days post-infection) 1.69 b 2.25 a 1.76 b days (8-14 days post-infection) 1.68 b 1.83 a 1.61 c days (0-14 days post-infection) 1.68 b 1.99 a 1.67 b days (Overall mean) 1.63 b 1.75 a 1.59 c Haemoglobin 6 DPI 9.85 a 5.55 c 7.97 b DPI Packed cell volume 6 DPI a c b DPI *Means bearing different superscripts in a row differ significantly Group I: Uninfected untreated control; Group II: Eimeria Spp. infected untreated control; Group III: Eimeria Spp. infected and vitamin E-Se with organic chromium treated Table 2: Effect of herbal vitamin E-selenium with organic chromium on parasitological parameters of broilers chicken infected with mixed Eimeria spp. Parameter Group I Group II t-value % Faecal score 0.00 b a ± % Survival 80.0 ± ± % Weight gain b ± a ± Performance index b ± a ± Average oocyst count/g a ± b ± % Reduction in oocyst production 0.00 b a ± Average oocyst index 4.0 a ± b ± Average lesion score 4.0 a ± b ± % Protection against lesion 0.0 b a ± *Means bearing different superscripts in a row differ significantly (P 0.05) Group I: Eimeria Spp. infected untreated control; Group II: Eimeria Spp. infected and vitamin E-Se with organic chromium treated 28

5 Selenium and vitamin E have also been shown to enhance the proliferation response of lymphocytes to mitogens thereby protecting host from the pathogenic effects of the parasite. Organic chromium used in the present study is also known to have antioxidant role (Tezuka et al., 1991). Due to potent antioxidant action it was also used in the study together with vitamin E-selenium so as to get better response and such response was actually found. Conclusion It can be suggested that the combination of herbal vitamin E-selenium with organic References Allen PC, Danforth HD and Augustine PC (1998). Dietary modulation of avian coccidiosis. International Journal of Parasitology, 28: Choct M, Naylor A J and Reinke N (2004). Selenium supplementation affect broiler growth performance, meat yield and feather coverage. British Poultry Science, 45: Colnago GL, Jensen LS and Long PL (1984). Effect of selenium and Vitamin E on development of immunity to coccidiosis in chickens. Poultry Science, 63: Davies SFM, Joyner LP and Kendall SB (1963). Coccidiosis. 1 st ed. Oliver and Boyd Ltd., Edinburg, p Hilbrich P (1978). Krankheiten des Geflugels unter besonderer berucksichtigung der Haltung and Futterung. Hermann Kuhn KG, Schwenningenam Necar, Germany. Hossain SM, Barreto SL and Silva CG (1998). Growth performance and carcass composition of broiler fed supplemental chromium from chromium yeast. Animal Feed Science Technique, 71: Johnson J and Reid NM (1970). Anticoccidial drugs: Lesion scoring techniques in battery and floor pen experiments with chickens. Experimental Parasitology, 28: Kidd M T (2004). Nutritional modulation of immune function in broilers. Poultry Science, 83: Lee DN, Wu FY, Cheng YS, Lin RS and Wu PC (2003). Effects of dietary chromium picolinate supplementation on growth performance and immune response of broilers. Asian-Australian Journal of Animal Sciences, 16: Lohakare JD, Hahn TW, Shim YH, Choi JY and Chae BJ (2004). Effect of feeding methods (Feed vs. chromium has the potential to provide protection in broiler chickens against coccidiosis induced alteration in growth, haematological and parasitological parameters. Acknowledgements The authors are thankful to M/s. Vetcare India Ltd., Bangalore for providing samples and financial support for this study and Dean, College of Veterinary and Animal Sciences, G.B. Pant University of Agriculture and Technology, Pantnagar, Uttarakhand, India for providing necessary facilities during the course of the study. Water) of vitamin E on growth performance and meat quality of broilers. Asian-Australian Journal of Animal Sciences, 17: Marsh JA, Dietert RR and Combs GF Jr (1982). Effect of dietary deficiencies of Vitamin E and selenium on primary lymphoid organs of the chick. Federation Proceeding, 41: 341. Morehouse NF and Baron RR (1970). Coccidiosis: Evaluation of coccidiosis by mortality, weight gains and faecal scores. Experimental Parasitology, 28: Natt MP and Herrick CA (1955). The effect of caecal coccidiosis on the blood cells of the domestic fowl. The changes in the blood volume during the course of infection. Poultry Science, 34: Nayak DC and Rai P (1985). Hemogram of chickens experimentally infected with Eimeria species. Indian Journal of Veterinary Medicine, 5: NRC (1997). The Role of Chromium in Animal Nutrition. National Academy Press, Washington, D.C. Sahin K, Kuut O and Sahin N (2001). Effects of dietary chromium picolinate supplementation on performance and plasma concentrations of insulin and corticosterene in laying hens under low ambient temperature..journal of Animal Physiology and Animal Nutrition (Berlin), 85: Schalm DW (1965). Veterinary Haematology II edn., Lea and Febiger, Philadelphia, pp Singh VS, Yadav CL, Palod J, Shukla PK and Kumar RR (2008). Effect of herbal vitamin E-Se supplement as an anticoccidial and growth promoter in broilers infected with mixed Eimeria species. Indian Journal of Animal physiology, 1:

6 Snedecor GW and Cochran WG (1994). Statistical Methods. VIII edn. Iowa University Press, Iowa, U.S.A. Tezuka M, Momiyama K, Edana T and Okada S (1991). Protective effect of chromium (III) on Distributing Company, Lucknow. acute lethal toxicity of carbon tetrachloride in rats and mice. Journal of Inorganic Biochemistry, 42: 1-8. Vegad J L Poultry Diseases, International Book 30