Eimeria tenella infected chickens and affected with tribasic copper ...

M. ANISIMOVA et a l...

EIMERIA TENELLA INFECTED CHICKENS AND AFFECTED WITH ...

Eimeria tenella infected chickens and affected with tribasic copper chloride M. Anisimova1, V.Koinarski2, M. Gabrashanska1, I. Vladov1, V. Ermakov3

INTRODUCTION It is known that helminthological status of animals depends on the content of heavy metals in the environment and feed [12]. But in some cases, salts of heavy metals (copper) are effective therapeutic agents at some parasitic infestations. This applies to birds helminthoses. Avian eimeriosis is one of the most common diseases in countries with industrial poultry bre eding. One of the most frequently and pathogenic protozoas in chicks is Eimeria tenella. Its pat hogenic influence on avian organism has been stu died in various aspects. Metabolic disorders take part place in the pathogenesis of eimeriosis [9]. Reduction of some small molecular weight anti oxidants (vitamin A, C and E) was established in chick eimeriosis by [3, 7].

Scientific paper UDC:636.52/. 58

animal growth, improved rate of gain and/or feed gain; superior blending in feed mixtures, and reduced handing costs. It has been widely used in feed formulations for most species, including chickens, pigs, beef and horses [11]. The following study was conducted to investi gate the interactive effects of TBCC and E. tenella infection on growth, feed intake, parasite status, liver copper and vitamin E concentration, as well as MDA level in chickens. MATERIALS AND METHODS

The study was performed on 60 clinically healthy broiler chickens, aged 20 days, weighted 335-370 g. Up to 11 days, the birds were housed in cages, under conditions excluding further Eimeria infection. At 11 days experimental groups of 20 birds each were established. The 1st group The essentiality of copper for poultry is well comprised healthy, untreated and un-infected birds. known [4]. Copper from various compounds has The birds from gr, 2 and 3 were infected once with often been added to poultry diets. L u o et al. [10] 8.104 sporulated E. tenella oocysts on day 13. The reported that the copper in tribasic copper chloride, fodder chickens from gr. 3 was supplemented with Cu(OH)3CI, is more available and save to broilers 400 mg Cu2+/kg food for 7 days - 2 days before than of copper sulfate when supplemented to diets. and 5 days post the inoculation. The experiment Because of its high water solubility and thus lasted 7 days p.i. On day 1 four chicks were killed hydroscopicity CuS04 leads to destructive reac to establish the level of vitamin E and Cu. On day 7 tions in feed mixtures. There are notoriously de birds were killed and the content of copper and structive in hot, humid climates [13]. Recognition vitamin E was determined. The level of Cu was es that TBCC would reduce feed stability problems tablished by atomic absorption spectrophotometry. lead to issuance of patents on the use of the The vitamin E content was established by HPLC compound as a nutritional source. method, using a fluorescence detector as descryCompared to CuS04, TBCC provides many be bed by Bieri et al. [2]. The total amount of lipid nefits including improved feed stability, less oxida peroxidation (LPO) product in plasma was asses tive destruction of vitamins and other essential feed sed using thiobarbituric acid (TBA) methods, mea ingredients; better copper bioavailability, improved suring spectrophotometrically MDA reactive pro ducts [14]. Body weight gain, feed intake and liver Cu and vitamin E were checked on day 1 and 7 p.i. Author's address: 11nsof Experimental Lesion scores and oocyst index were checked on Morphology, Pathology and Anthropology with day 7. Museum, BAS 1113 Sofia, Bulgaria, 2Faculty of Veterinary Medicine, Trakia University, 6000 Stara RESULTS AND DISCUSSION Zagora, Bulgaria, 3Vernadsky Institute of Geoche mistry and Analytical Chemistry of RAS, 11991 Figure 1 and 2 presented the live weight gain Moscow, Russia and the feed conversion ratio (FCR) in chickens Paper received: 03. 10. 2013. infected and uninfected with tenella and infected

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with E. tenella and supplemented with copper. The results evidenced that at the beginning of the experiment, the body weight in all groups was equal but at end, it was significantly lower in the infected birds (gr.2) compared to the healthy chickens. This decrease was obviously due to the relatively low body weight gain in all infected birds compared to healthy controls.

by chickens infected with , demonstrating the development of infection. It was find that the oocyst index and lesion scores in infected and Cu supplemented 20-day-old chickens significant decreased, compared to the birds in gr. 2. Figure 4 presents Cu level. It was reduced in the infected chickens compared to the healthy ones. Cu con centration was increased after Cu supplementation. The liver and plasma vitamin E levels are presented in figure 5. TBCC increased significantly liver vitamin E content in the infected with E. tenella birds compared with only infected ones. 7/2

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Figure 1 - Body weight gain in healthy and E. tenella infected broiler-chickens

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Lesion scores

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h-i Figure 3 - Lesions scores and the oocyst index in healthy and E. tenella broiler-chickens

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No. group Figure 2 - Feed conversion ratio in healthy and E. tenella infected broiler-chickens After Cu supplementation weight gain was found to significantly higher, compared to infected un-supplemented birds. At the same time, FCR was higher in the infected chickens, compared to the controls. After supplemented with TBCC, it was found that FCR to be low, compared to infected un supplemented birds. Figure 3. presents the lesion scores and the oocyst index. Our results showed that 7 days, post challenge, E. tenella produced substantial injures in the caeca manifested by visible erosions of the mucosa, extremely tinned mucosa and blood on the intestinal lumen as well as enlargement in the size of the intestine. The oocyst index in infected but non-treated birds as expected, indicated that oocysts had been released in significant amounts

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No. group Figure 4 - Liver copper levels in healthy and E. tenella infected broiler-chickens Figure 6 presented the level of MDA. It was significantly increased in chickens infected with eimeriae (supplemented and non-supplemented) compared with healthy controls.




Figure 5 - Liver and plasma vitamin E concentration in healthy and E. tenella infected broiler-chickens. Figure 6 presented the level of MDA. It was significantly increased in chickens infected with eimeriae (supplemented and non-supplemented) compared with healthy controls. In this study while the level of MDA showed an increase, indicated that E. tenella led to lipid peroxidation during the specified period and decreased in the antioxidant vitamin E, which was involved in the non-enzymatic mechanisms, resulted in the occurrence of oxidative damage. There are several data that the parasitic infections may lead to a decrease in the blood level of antioxidant vitamins. E. tenella oocysts provoked the lipid peroxidation at the LPO at the specific period and the quantity.

Figure 6 - Plasma MDA concentration in healthy and E. tenella infected broiler-chickens. The occurrence of oxidative damage may be contributed directly to the mechanism of negative effects of the disease. As shown by us vitamin E

was significantly depressed by the infection with tenella, but increased with dietary Cu supple mentation. Allen and Fetterer [1] showed that vita min E was reduced in infection with E. maxima. This reduction could be due to vitamin E malab sorption in mid-intestine, which is parasitized by E. maxima. But E. tenella infects the ceca and is not a major effector of lipid malabsorption, but is asso ciated with an increase of nitric oxide species [15] and a decrease antioxidant enzyme SOD [7]. Depressed appetite and the bleeding observed in E. tenella infection as well as increased tissue demand for vitamin E to neutralize free radicals may account for the decreased plasma vitamin E in our experiment. Our data showed that the erythro cyte concentration of MDA was significantly higher in infected chicken than healthy ones. MDA is used a biomarker for radical- induced damages of biolo gical membranes [5]. The higher level of MDA could be attributed to lipid peroxidation resulting from increased reactive-oxygen species (ROS). Cu supplementation applied in physiological norm affe cted FOR and body weight gain positively. Body weight gain was similar in the infected, supple mented and controls. The lack of a detrimental effect on body weight by TBCC supplementation in chickens was established by [10]. The authors showed positive effect of TBCC on broiler performance, relative copper bioavailability and oxidation stability of vitamin E. Vitamin E was increased after TBCC supplementation in broilers. Liver Cu concentration was elevated in chickens consuming diets that contained from 150-450 mg/kg supplemented copper [11]. Increased Cu level could influence positively the activity of Cu, Zn superoxide dismutase, acting as an antioxidant enzyme [6]. So that both antioxidants (Cu,Zn-SOD and vitamin E) were increased in the liver in chickens challenged with E. tenella [8] and treated with copper. In that case TBCC was increased antioxidant defense system in the chickens infected with E. tenella. Our data with regard to the lesion scores and the oocysts index value indicated that by day 7 p.i. E. tenella provoked significant injuries to the intestinal mucosa. Alterations in the cecum and oocyst production were indicative of a severe infection involving pathogenic oxidative stress. The established changes in caeca and the oocyst production in the birds in gr. 3 were indicative of beneficial effect of TBCC on the lesions scores and the oocyst index on eimeriosis infection. Combined effect of Cu and the infection on the lesion scores and oocyst indexes was significantly presented. They were reduced probably due to Cu supple mentation.


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CONCLUSION

tenella in broiler chickens// Vet J, 2006. Vol 172 No. 3. P. 488-492. TBCC possesses several chemical characterristics that make it desirable as a Cu source in [8] Hooge D., Steward F., J. McNaufhton. Improved production, such as small particle size, excellent stabilities of vitamin A, D3, E and riboflavin with tribasic copper chloride (TBCC) compared to flow, and low water solubility (Miles et al., 1998). It copper sulfate pentahydrate in crumbled broiler was established that TBCC was less active than starter feed HP oultry Sci., 2000. Vol. 79 (suppl 1) copper sulfate in promoting the oxidative stress [8, P. 43-44. 9]. TBCC could be used in poultry as additive compound during eimeriosis. Our results are [9] Koinarski V., Pavlov A. Studies on the meat com suggestive of the beneficial effects of TBCC on bird position in chickens invaded with Eimeria tenella// Bulg. J. Vet Med., 2001. Vol. 4. P. 243-248. performance and the prevention of lesions and oocysts caused by E.tenella. The data [10] Luo from X.G., this Ji F., Lin Y.X., F. Steward A., Lu L., Liu experiment indicate that TBCC is a safe product B. and Yu S.X. Effects of dietary supplementation and well available to broiler challenged with E. with copper sulfate or tribasic copper chloride on tenella without any negative indices. broiler performance, relative copper bioavailability, and oxidation stability of vitamin E Acknowledgement in feed// Poult. Sci., 2005. Vol. 84. P. 888-893. This study was supported by a bilateral project [11] Miles R., O'Keefe, Henry P., Lewis A., C. between Bulgarian and Russian Academy o f Anmmerman, Luo X. The effect of dietary Sciences. supplementation with copper sulfate or tribasic copper chloride on broiler performance, relative REFERENCES copper bioavailability and dietary prooxidant [1] Allen P.C., Fetterer R.H. Recent advances in bio activity// Poultry Sci., 1998. Vol. 77. P. 416-425. logy and immunobiology of Eimeria species and [12] Nanev V, Habrashanska M., Ermakov V., Tjutikov in diagnosis and control of infection with these S. Heavy metals and helmintological status in coccidian parasites of poultry// Clinical Micro fallow deer from Bulgaria// Ecologica, 2010. Vol biology Reviews, 2002. Vol. 15. No. 1. P. 58. 17. No. 58. P. 113-116. [2] Bieri J., Tolver T., Catigan G. Simultaneous deter [13] Palache A., Berman H„ Fronde C. Halides, nitra mination of alfa-tocopherol and retinol in plasma tes, borates, carbonates, sulfates, phosphates, or red blood cells by high pressure liquid chroarsenates, tungstates, molybdates, etc. //John matography//Lipids, 1981. Vol. 16. P. 777-779. Wileyand Sons. The system of Mineralogy of [3] Coles B., Blely J., March B. Vitamin A deficiency James Dwight Dana and Esward Salisbury Dana. and Eimeria acen/ulina infection in the chicks// 7th ed. Vol. II. Jale University, New York, 1994 P Poultry Sci., 1974. Vol. 90. P. 1295-301. 1837-1892. [4] Davis G., Mertz W. Copper: Trace Elements in [14] Placer Z., Cushman L., Johnson B. Estimation of Human and Animal Nutrition. Vol.1. W. Merts ed. product of lipid peroxidation (malondialdehyde) in Academic Press, New York, 1974. P. 301-304. biochemical systems//Anal. Biochem., 1966. Vol [5] Day C. P. Is necroinflammation a prerequisite for 16. P. 359-364. fibrosis?// Hepatogastroenterology, 1996. Vol 43 [15] Wang S„ Zhang J., Zhang Y„ Kern S„ Danner P. 104-120. R.L. (2008). Nitric oxide-p38 МАРК signaling [6] Fridovich I. Supeoxide dismutase// Ann. Review stabilizes mRNA through AU-rich elementof Biochem, 1975. Vol. 44. P. 147-159. dependent and -independent mechanisms// J [7] Georgieva N„ Koinarski V., Gadjeva V. Antioxi Leukoc Biol., 2008. Vol. 83. P. 982-990. dant status during the course of an Eimeria

ABSTRACT EIMERIA TENELLA INFECTED CHICKENS AND AFFECTED WITH TRIBASIC COPPER CHLORIDE The effect of tribasic copper chloride (TBCC) on broiler chickens experimentally infected with Eimeria tenella (Protozoa) was investigated. Body performance, parasite burden, liver vitamin E, liver copper and plasma malondialdehyde level were measured. The results showed improved body weight gain, reduced feed conversion ratio, elevated liver vitamin E and Cu content, as well as increased MDA content. The data from this experiment indicate that TBCC is a safe product and well available to broiler challenged with E. tenella without any negative indices. Keywords: TBCC, Eimeria tenella, vitamin E, copper, broiler performance

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