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

Immunization of lambs against infection with echinucoccus granulosus using protoscolices and hydatid fluid antigens

In the present study, protoscolices and hydatid fluid were prepared from livers or lungs of sheep with hydatid cyst in sterile conditions. The protein concentration of samples was then measured by Bradford method. 12 lambs 4 - 6 months of age of mixed sex were randomly allocated to 3 groups of 4 lambs. Each lamb in groups 1 and 2 was immunized subcutaneously in the neck with a 2 - ml dose of vaccine [1 mg of protoscolex and hydatid fluid proteins dissolved in 1 ml of PBS] and emulsified with an equal volume of Freund's complete adjuvant [FCA] respectively. Control lambs were immunized with adjuvant in PBS. Lambs were boosted at day 28 with the same preparation 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 10 adult E. granulosus. All lambs were euthanized after 7 months and examined for hydatid cysts. Results showed that, the number of cysts in immunized lambs with protoscolices were lower than in control group and also the number of cysts in immunized lambs with hydatid fluid antigen was significant [p < 0.05]. This result indicated that the protective immunity in lambs with protoscolices and hydatid fluid was 54.5% and 75.75% respectively

[Isolation and identification of facultative aerobic-anaerobic gram-positive bacteria in hatching eggs]

To detect and isolate the Gram-positive aerobic and facultative- anaerobic bacteria in hatching eggs. Prospective longitudinal study. One hundred and twenty eggs from the broiler breeder stock and hatchery and 30 new-hatched chicks. Bacteiological studies were done in five different developing stages: immediatley after laying, before and after disinfecting, before moving from setter to hatcher and from new hatched chicks. Furthermore, two different samples were taken from the shell and the yolk of each egg. Eggs were washed with Nutrient-Broth Media, sotred in 37[degree sign] C for 24 hours and finally transferred to blood agar containing potassium telorite. Yolk samples were directly transferred to blood agar containing potassium telorite. This process was repeated similarly for those samples from yolk sac of 18-day-old embryos and newborn chicks. Exact fisher test. The contamination rate of eggshells in different stages were 96.66%, 100%, 100% and 90%, respectively. Bacillus cereus, B. subtilis, B. polymixa and B. coagulans were isolated in 44.15%, 64.16%, 7.4% and 2.5% of cases, respectively. Staphylococcus aureus and S. saprophyticus were isolated from one yolk [3.3%] and tow yolk sacs [6.6%]. Any bacteria was isolated from yolk sac of 18 - day- old embryos. The source of such contamination is mostly related to eggshells. In spite of routine managing programs in breeder stocks[ such as selection, cleaning, and disinfection of eggs] it dose not effect to reduce Gram positive bacteria on egg shells and contamination of eggs in setter. Hence, yolk sac infection occurs and causes mortality in chicks

Detection of hydatid fluid and protoscolices antigens in sheep with hydatidosis

Hydatidosis is a disease caused by infection with the metacestode stage of the dog tapeworm, Echinococcus granulosus. This is recognised as one of the world's most important zoonoses, affecting both humans and their domestic animals. In the current study, 20 sheep cystic livers or lungs were collected. Hydatid fluid and protoscolices were isolated and the soluble protein was prepared. Polypeptide profile of hydatid fluid and protoscolices was analysed by SDS-PAGE with 12.5% acrylamide concentration. Humoral immunity and antigenic pattern were evaluated by Western blotting. In electrophoresis of hydatid fluid, five bands with molecular weight of 84, 68-70, 55, 27-28 and 16-17 kDa were observed; the 68-70 and 55 kDa bands were noticeably dominant. In Western blot of hydatid fluid protein, only 68-70 and 55 kDa bands had conciderable positive reaction. A total of 12 bands were also observed in protoscolices polypeptide profile. The molecular weight of the bands were 120, 88-89, 84, 66, 55-56, 49-50, 43-44, 36, 30-31, 24, 20 and 15- 16 kDa. Seven bands with molecular weight of 66, 55-56, 49-50, 43-44, 36, 30-31 and 24 kDa had positive reaction in Western blot We concluded that the determination of specific antigenic bands for sheep hydatid fluid and protoscolices was successfully achieved in this study

Immunization of rabbits against Hyalomma anatolicum anatolicum tick using antigens derived from whole tick and gut extract

This study was conducted in three phases for identification of protective antigens in Hyalomma anatolicum anatolicum that can induce the best protection in the sheep and cattle. In the first phase of study, H. anatolicum anatolicum was cultured for preparing enough antigens. Then, four antigenic preparation, viz, whole tick supernatant antigen [WTSA], whole tick pellet antigen [WTPA], gut supernatant antigen [GSA] and gut pellet antigen [GPA] were made partially fed adult female H. anatolicum anatolicum. Four groups of five rabbits were immunized with the antigens in Freund's adjuvant emulsion and a fifth group was kept as control. The rabbits were challenged with ten pairs of homologus ticks and characteristic representing tick feeding and fertility were recorded and compared between groups. A significant decrease in percent of engorgement, index feeding, percent of oviposited, weight of egg and index fertility were observed in GSA immunized rabbits