Molecular detection of equine piroplasms in donkeys [equus asinus] in North Khorasan province, Iran

Equine piroplasmosis is a tickborne disease of equids with worldwide distribution, caused by Theileria equi and Babesia caballi. The aim of this study was molecular detection of T. equi and B. caballi in donkeys in northeastern Iran and investigate the association between positivity of piroplasm infection and host-related factors. In the present study, Blood samples were collected from 106 apparently healthy donkeys [Equus asinus] in North Khorasan province, Iran. Blood smears were prepared and stained by giemsa method. DNA was extracted from blood and then multiplex-PCR was done for detection of any piroplasms infection. According to the results, four donkeys showed T. equi in blood smears but B. caballi was not found. Also, fifty four donkeys [50.94%] showed T. equi infection using multiplex-PCR. No siginificant difference was observed between the frequency of T. equi infection with hostrelated factors in donkeys. This is the first report on the molecular detection of eqiune piroplamosis in donkeys in Iran. Also, no significant association was found between the rate of T. equi infected animals

Molecular detection of Babesia spp in sheep and vector ticks in North Khorasan province, Iran

Babesiosis is an important tickborne disease in the sheep of Iran. A molecular study was carried out in North Khorasan province, Iran in 2010-2011, designed to identify Babesia spp. infection of both sheep and ticks. Ninety sheep from different flocks were clinically examined and blood samples were collected with ixodid ticks. The collected ticks were separated into 82 tick pools and the salivary glands were dissected out in 0.85% [w/v] saline under a stereomicroscope. The blood and the salivary glands were examined using semi-nested PCR. Piroplasm infection was detected in 37 blood smears using microcopic examination while 80 blood samples were piroplasm positive in the first round of semi-nested PCR and Babesia ovis was only detected in 6 [6.6%] of positive samples in the second round of semi-nested PCR. Of the 434 ticks that were collected, the most prevalent species was Rhipicephalus turanicus [69.3%] followed by Hyalomma marginatum turanicum [18.4%], Dermacentor marginatus [6.4%] and Rhipicephalus bursa [5.7%]. One pool of H. m. turanicum salivary glands and one pool of R.turanicus were infected with B. ovis. Based on these results, it is concluded that B.ovis has a low prevalence among the sheep of North Khorasan province and H. m. turanicum and R. turanicus may be the vectors of B.ovis in this area

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

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%

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