Genetic diversity of bovine Neospora caninum determined by microsatellite markers.

Neospora caninum is one of the most significant parasitic organisms causing bovine abortion worldwide. Despite the economic impact of this infection, relatively little is known about the genetic diversity of this parasite. In this study, using Nc5 and ITS1 nested PCR, N. caninum has been detected in 12 brain samples of aborted fetuses from 298 seropositive dairy cattle collected from four different regions in Tehran, Iran. These specimen (Nc-Iran) were genotyped in multilocus using 9 different microsatellite markers previously described (MS4, MS5, MS6A, MS6B, MS7, MS8, MS10, MS12 and MS21). Microsatellite amplification was completely feasible in 2 samples, semi-completely in 8 samples, and failed in 2 samples. Within the two completely performed allelic profiles of Nc-Iran strains, unique multilocus profiles were obtained for both and novel allelic patterns were found in the MS8 and MS10 microsatellite markers. The Jaccard's similarity index showed significant difference between these two strains and from other standard isolates derived from GenBank such as Nc-Liv, Nc-SweB1, Nc-GER1, KBA1, and KBA2. All samples originating from the same area showed identical allelic numbers and a correlation between the number of repeats and geographic districts was observed.

Molecular identification of ovine Theileria species by a new PCR-RFLP method.

Theileria spp. infect wild and domestic ruminants in the tropical and subtropical regions of the world. Two species, T. lestoquardi and T. ovis, are suspected to cause ovine theileriosis in Iran. The epidemiological aspects of ovine theileriosis in Iran are poorly understood, and further investigations by sensitive and precise techniques are required. In this study, the use of a nested PCR for amplification of a fragment of the 18S ribosomal DNA from virtually all species of Theileria is described. For differentiation of various Theileria spp. a RFLP assay was developed as a diagnostic tool enabling direct, concurrent, highly specific and sensitive identification of Theileria spp. The sensitivity of the nested PCR for Theileria species was 10(-5)% parasitemia. Restriction fragment length polymorphism (RFLP) of the PCR products allowed differentiation between three different Theileria species (T. annulata, T. lestoquardi and T. ovis) and seems to be useful for differentiation of other species such as T. separata and Theileria spp. china. From 100 field blood samples obtained from sheep in East and South-East Iran, 56% were positive for Theileria spp. by nested-PCR compared with 21% by microscopic examination. Out of 56 positive samples, 12.5% (7/56) were positive for T. ovis and 87/5% (49/56) were positive for T. lestoquardi. This is the first report in which T. ovis has been detected in Iran using molecular identification techniques.

Seroprevalence of Neospora caninum infection in dogs from rural and urban environments in Tehran, Iran.

103 serum samples were taken from owned dogs in Tehran (53 urban dogs and 50 farm dogs) from February to May 2005 and tested for IgG antibody against Neospora caninum using the immunofluorescence antibody test. Urban dogs were selected from apparent healthy dogs referred to the Small Animal Hospital of Faculty of Veterinary Medicine, University of Tehran, for routine operation, and the farm dogs were selected from the sheepdogs in sheep flocks around Tehran randomly. The titer of 1:50 was selected as cutoff. In total, 20 of 103 cases (19.4%) were identified as positive. The infection rate in farm dogs (28%) was higher than in urban dogs (11.3%), and the difference was statistically significant (p < 0.05). The infection rates in different genders and age groups were determined. The differences between seroprevalence of N. caninum infection in different sexes were not significant. A high correlation was observed between the infection rate and age (r (2) = 0.97, p < 0.01).

Seroprevalence of Toxoplasma gondii infection in stray and household cats in Tehran.

The prevalence of anti-Toxoplasma gondii specific IgG in stray and household cats in Tehran was determined by Indirect Fluorescent Antibody Test (IFAT) on serum samples from 100 cats (50 stray and 50 households). Overall infection rate was 63%. The infection rate in stray cats (90%) was significantly higher (p<0.001) than that of household cats (36%). Last serum positive dilutions varied from 1: 32 to 1: 512 titres in which the highest percentage (27%) was for 1:256 and the least (4.8%) was at 1:32. The rate of infection between male and female cats of both groups was not significantly different; 90.3% versus 89.5% for male and female in stray cats, respectively. Different sexes of household cats were seropositive at the same rate (36%). A high positive correlation (r(2)=0.97) between age and the rate of infection was observed. The prevalence of Toxoplasma gondii infection in cats in Tehran was high, especially in stray cats which are probably the main source of Toxoplasma infection in this area.