Antioxidant and anticoccidial effects of garlic powder and sulfur amino acids on Eimeria-infected and uninfected broiler chickens

To study the anticoccidial and antioxidant effects of garlic powder [GP] and total sulfur amino acid [TSAA] on growth performance, faecal oocyst output, oxidative stress indicators and antioxidant status markers in broiler chickens, a 2 × 2 × 2 splitplot- factorial arrangement of treatments was used. A total of 480-day-old male broiler chickens were equally assigned to two plots. Two hundred forty chickens were challenged with Eimeria oocysts species mix by oral inoculation at day 34 [infected plot] and the other half was left unchallenged. In each plot, chickens were r and omly assigned to four treatments with 5 replicates of 12 birds each, and fed one of the following diets: basal diet, basal diet plus 0.5% GP, basal diet plus 50% more TSAA, basal diet plus 0.5% GP and 50% more TSAA. The results showed that inoculation with 7.5 × 102 oocysts of Eimeria mixed species significantly reduced growth performance [P<0.05] and increased nitric oxide [NO] and malonyldialdehyde [MDA], but did not change the hepatic activity of superoxide dismutase [SOD] and the glutathione peroxidase [GPX] activity. Growth depression and increased NO and MDA were greater in infected than uninfected birds. In the infected birds fed with the basal diet without any supplementation, NO and MDA were significantly greater [P<0.05] in comparison with other groups. Faecal oocyst output significantly decreased with the supplementation of GP and TSAA in infected birds [P<0.05]. In conclusion, the supplementation of GP and 50% TSAA individually or in combination with the basal diet significantly improved ADG [P<0.05] and decreased OPG, MDAandNO. Moreover, no changes in the antioxidant enzymes were observed in birds infected with Eimeria. Key words: Coccidiosis, Broiler chickens, Oxidative stress, Garlic powder, Sulfurs amino acid

Evaluation of Echinococcus granulosus coproantigens by Dot-lotting in dogs

Definitive hosts of the Echinococcus granulosus [E. granulosus] parasite are carnivores such as dogs, wolves and foxes. Detection of this parasite through faecal examination is not possible. In this study, dot-blotting test for E. granulosus-specific coproantigens has been evaluated in dog. Three 2-3-month-old puppies were treated with piperazine and then faecal samples were collected as pre-infection samples. Seven days later, hydatid cysts from livers and lungs of sheep were fed to the puppies. Faecal samples were collected weekly for five weeks as post-infection samples. Soluble protein of pre- and post-infection faecal samples were prepared and dot-blotting test was conducted. In parallel experiments, the presence of E. granulosus eggs and also dot-blotting test were evaluated in 15 faecal samples of dogs collected from Razi Veterinary Hospital in Mashhad. For the detection of protein bands in pre-infection and fifth-week post-infection samples, polypeptide profile was analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis [SDS-PAGE]. The results showed that incremental spot colours was observed in samples of experimentally infected dogs collected from the first to fifth post-infection weeks. In dot-blot analysis of faecal samples in 15 dogs, 4 samples were positive, and also these four samples were positive for E. granulosus eggs. In SDS-PAGE, one band in pre-infection and four bands in fifth-week post-infection samples were observed. The molecular weight of pre-infection sample of experimentally infected dogs was 16 kDa and the molecular weights of the samples collected five weeks post-infection were 14, 22, 36 and 45 kDa, respectively. In conclusion, the results of this experiment showed that the dot-blotting method does not produce a reliable outcome. For evaluation of the specific coproantigens of E. granulosus in dogs, coproantigen-ELISA test is needed

Antibody response against hydatid fluid, protoscolex and whole body of Echinococcus granulosus antigens in lambs

In this study, the immunogenicity of three types of antigens [hydatid fluid, protoscolices and whole body of E. granulosus] was investigated in lambs by ELISA. Sixteen 4-6-month-old lambs of mixed sexes were divided into 4 groups of 4 lambs [three immunized and one control group]. Twelve lambs as immunized groups received 2 mg of hydatid fluid, protoscolices and whole body of E. granulosus antigens dissolved in 1 ml of PBS per immunization for each lamb, respectively. As an adjuvant, Freund's complete adjuvant [FCA] was mixed with antigens to form a water-in-oil emulsion which was inoculated subcutaneously on day 1 of the trial. Each control lamb was inoculated with a total of 2 ml of PBS emulsified in equal volumes of FCA. Lambs were boosted on day 28 with the same preparation as described above except that FCA was replaced by Freund's incomplete adjuvant [FIA]. Three weeks after the second immunization, each lamb received a challenge infection with 2000 protoscolices intraperitoneally and also 10 gravid proglotid of E. granulosus orally. Sera were collected before and after immunization and serum antibodies were tested by ELISA. The results showed that the production of antibody had a significant difference between the test groups and the control [P<0.05]. Lambs immunized with whole body of E. granulosus showed the highest antibody production. The level of antibody production between the lambs immunized with hydatid fluid and the protoscolices was not different significantly [P>0.05], whereas, the level of antibody production between the lambs immunized with hydatid fluid and whole body of E. granulosus was different significantly [P<0.05]. The results of this study showed that the antigens of whole body of E. granulosus might be a good candidate for immunization and diagnosis of hydatid cyst in the intermediate hosts of E. granulosus

Immunization of rabbits against Hyalomma anatolicum anatolicum using larval and nymphal extracts

The protective capacity of the crude extracts from larval and nymphal stages of Hyalomma anatolicum anatolicum was examined in the New Zealand white rabbits. The rabbits were randomly divided into 3 groups of five animals. The rabbits in groups 1 and 2 were immunized with crude extracts of larval and nymphal antigens, respectively and group 3 was served as control. Following challenge of each rabbit with 2000 larvae of H. a. anatolicum, engorged nymphs were collected, weighed and then cultured in an incubator. A significant decrease in weight of engorged nymphs was only observed in group 1 that immunized with crude extracts of tick's larvae [P<0.05]. Polypeptide profile of the larval and nymphal extracts was analysed by SDS-PAGE, and antigenic pattern with serum of immunized rabbits was evaluated by Dot and Western blot test. The molecular weight of the fraction of larval extract after SDS-PAGE showed six polypeptide bands as follows: 97, 84, 66, 55, 45 and 36 kDa, and in nymphal extract 13 polypeptide bands of 205, 116, 97, 84, 66, 55, 45, 36, 29, 24 and 20 kDa and two bands between 116 and 205 kDa were found. In Western blotting, positive reaction was only observed with sera of group 1 in the bands with 97, 84 and 66 kDa. It seems that the larval extract of H. a. anatolicum can be used as a source of biological material for isolation of protective antigen

Detection of midgut antigens of Hyalomma anatolicum anatolicum tick using SDS-PAGE and Western blot

Ticks are important ectoparasites which are a considerable threat to human beings and to animals all over the world. Enormous economic losses annually occur in livestock production around the world as a result of their existance. One of the ways to control ticks and tick-borne diseases is to introduce resistance to these ectoparasites through immunization. For identification of the putative protective antigens, screening of large number of parasite antigens and their fractions are necessary. In this study, midguts of fed adult female Hyalomma anatolicum anatolicum were used to prepare antigen and to identify the midgut profile. Polypeptide profile was analysed by SDS-PAGE with 12.5% concentration under denaturated conditions and discontinuous buffer system. Humoral immunity and antigenic pattern were evaluated by Western blot. A total of 4 fractions were observed in the polypeptide profile. The molecular weight of the fractions were 97, 84, 66 and 55 kDa. The band with molecular weight of 66 kDa was dominant. Positive reaction with 84, 66 and 55 kDa bands were observed in immuno-blot of the midgut antigens

First report of the Hadjelia truncata infestation in pigeons of Iran

Three adult pigeons [Columbia livid] belonging to a pigeon farm in Mashhad area were submitted to Poultry Clinic of School of Veterinary Medicine for post-mortem inspection. The farm had 150 adult pigeons and a number of them suffered from a chronic disease with body weight loss, diarrhoea and weakness. At necropsy, slight to extreme enlargement with distortion were observed in gizzards of all carcases. There was a large number of nematodes in or beneath the lining of the affected gizzards. The worms were removed from the lining of the rostral parts of the gizzards and cleared in lactophenol. Also, tissue blocks from various parts of the gizzards were processed by conventional methods for preparation of paraffin wax sections. Based on parasitological and pathological findings the Hadjelia truncata infestation was identified for the first time from Iran and this species is a pathogenic agent for pigeons

Immunization of lambs with whole body Echinococcus granulosus

We conducted this study to determine the level of immunity after vaccination of lambs with whole body Echinococcus granulosus. To do so, 200 mature E. granulosus parasites, which were kept in 10% formaline for 8 months, were obtained from the Department of Parasitology, School of Veterinary Medicine, Ferdowsi University of Mashhad. The soluble protein of the parasite was prepared. The sample was homogenized in a blender, sonicated on ice and then centrifugated for 15 min at 10,000 g. Final yield was kept at -20°C until used. Eight 4-6-month-old lambs of mixed sex, were divided into 2 equal groups; each lamb in the test group was vaccinated subcutaneously in the neck with 2 ml of the vaccine [1 mg of whole body of E. granulosus protein dissolved in 1 ml of PBS plus 1 ml of Freund's complete adjuvant [FCA]]. The control lambs were vaccinated only with adjuvant in PBS. Lambs were re-vaccinated four weeks after the first vaccination with the same preparation except that FCA was replaced by Freund's incomplete adjuvant [FIA]. Three weeks later, each lamb was administered a challenge infection dose of 2000 protoscolices intraperitoneally and 10 adult E. granulosus. After 7 months, all lambs were killed and examined for hydatid cysts. We found two cysts in the liver and and one in the lung of only one of the vaccinated lambs. The number of cysts in vaccinated lambs were significantly lower than that in the control group [P<0.001]. This means that the protective immunity in lambs with whole body of E. granulosus was approximately 90%

[ case-report of an outbrebreak of histomoniasis in Turkey in mashhad area]

Diagnosis of Histomoniasis. Case-report. Eighty turkeys [2-3 months old] from a small flock in Mashhad area. In February 2003, in a small flock of turkeys [n=80] that co-bred with domestic hens [n=200] and geese [n=70] a disease occurred with clinical signs of inappetence, diarrhea and head cyanosis. Disease had high morbidity [75%] and mortality [66%]. Five carcases and two ill turkeys were refreed to birds clinic for diagnosis and tretment of disease. At necropsy, congestion and necrosis in mucosa of ceca and liver were characterized. Because of strong suspicion to histomoniasis, tissue samples were obtained for Parasitological and Pathological examinations. After preparing smears and tissue sections, they were stained with Giemsa and Haematoxyline-Eosine methods, respectively. Also, faeces of domestic hens were examined for determination Heterakis gallinarum infestation. Microscopical examination of the smears and tissue sections were shown Histomonas melagridis infection in cecum and liver. Also the hetrakid or ascarid eggs were found in the faeces. The results of this study indicates that histomoniasis can be an important disease of turkeys in Iran when co-bred with chicken