Ovine theileriosis is an important hemoprotozoal disease of sheep and goats in tropical and subtropical regions that leads to economic losses in these animals. A nested PCR-restriction fragment length polymorphism (RFLP) was carried out to identification Theileria species in sheep in some area in western half of Iran (Sari, Rasht, Urmia, Ilam, and Ahvaz). Two hundred and fifty blood samples were taken from sheep during tick activating season (summer of 2008). Microscopic examination revealed that 9.2% (23/250) sheep were infected by Theileria spp. piroplasms. Parasitemia ranged from 0.011% to 0.015%. In nested PCR assessment of DNA samples, 32.8% (82/250) sheep were positive. The negative samples were confirmed by amplifying of ovine beta-actin gene as an internal control. The differentiation of Theileria species was based on RFLP patterns using three restriction enzymes: HpaII, Rsa1, and Bsh 1285I. Out of 82 positive samples, 54.8% (45/82) and 40.2% (33/82) were positive for Theileria lestoquardi and Theileria ovis respectively. Mixed infection was detected in 4.8% (4/82) cases. Based on their PCR product digestion pattern with HpaII (1178, 900, 278, and 106 bp), it seemed to be mixture of Theileria annulata and T. lestoquardi. The presence of T. annulata was supported by sequence analysis. This is the first report of naturally infected sheep with T. annulata in Iran. Geographical distribution of Theileria species in sheep is shown according to the result of microscopy and nested PCR and RFLP data.

A few studies have been done on the seroepidemiology of anti-Neospora caninum antibodies in dairy and beef cattle farms in Iran, which suggested the presence of N. caninum in these areas, but there is no published information directed on the presence or epidemiology of this organism in the dogs in Iran. To investigate anti-N. caninum antibodies in household dogs and dogs living in cattle farms, 100 blood samples were collected: 50 from dogs living in dairy and beef cattle farms and 50 from household dogs. Serum samples were screened for detection of anti-N. caninum IgG antibodies using indirect fluorescent antibody test (IFAT; > or = 50). Antibodies were seen in 10 (20%) of 50 household dogs and in 23 (46%) of 50 farm dogs. There were significant statistical differences in seropositivity between these two groups (P = 0.005). The IFAT antibody titers were as follows: 1:50 in seven dogs, 1:100 in eight dogs, 1:200 in six dogs, 1:400 in seven dogs, 1:800 in three dogs, 1:1,600 in one dog, and 1:12,800 in one dog. There were no significant differences in seropositivity between males and females. The positive results were increasing with age, and positive results were significantly different in the age group of older than 2 years compared to the dogs of age group under 1 year (P = 0.000) and 1-2 years (P = 0.007). The results confirm the exposure of household and farm dogs to N. caninum in Tehran and the higher rate of exposure for the dogs of dairy and cattle farms around Tehran.

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Virulence of 11 native strains of entomopathogenic fungi; Metarhizium anisopliae (three strains), Beauveria bassiana (six strains) and Lecanicillium psalliotae (two strains) collected from different parts of Iran, were studied against different developmental stages of Rhipicephalus (Boophilus) annulatus. After the exposure of ticks to the fungal strains in different concentrations (i.e. 10(3), 10(5), 10(7) conidia/ml), various parameters such as mortality rate and reproductive efficiency of engorged females, mortality of unfed tick larvae and eclosion percentage of infected eggs were evaluated to determine the fungal virulence. Based on the obtained results, five strains including M. anisopliae (IRAN 437 C and DEMI 001), B. bassiana (IRAN 403 C) and L. psalliotae (IRAN 468 C and IRAN 518 C) were found to be virulent to various stages of tick developmental cycle. Mortality rate of engorged females was found to be dose-dependent with regard to the conidial concentration used. Total mortality rates of 90-100%, 70% and 56.6% were observed for M. anisopliae (IRAN 437 C and DEMI 001), B. bassiana (IRAN 403 C) and L. psalliotae (IRAN 468 C), 6-11 days post inoculation (PI) with 10(7) conidia/ml, respectively. Most strains were able to inhibit egg laying by females in the range of 0-26% in different conidial concentrations. The results indicated that the mean egg laying of treated engorged tick females exposed to M. anisopliae (IRAN 437 C) was less than the mean values of those treated with other fungal strains. Results revealed 89.1%, 35.5% and 56.3% decrease in egg hatchability and 88.69%, 78.15% and 59.74% reduction in reproductive efficiency of the ticks using 10(7) conidia/ml of M. anisopliae (IRAN 437 C), B. bassiana (IRAN 403 C) and L. psalliotae (IRAN 468 C), respectively. In general, the entomopathogenic effects of native M. anisopliae and B. bassiana against various developmental stages of R. (B.) annulatus were confirmed in the present work. Likewise, although L. psalliotae, which was introduced for the first time as an entomopathogenic fungus against tick had not more than 13.3% mortality effect against adult females, but its effect on egg hatchability and reproductive efficiency was remarkable.

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