[Prevalence and severity of animal Fasciolosis in six provinces of Iran]

Fasciolosis is one of the most important parasitic disease common among both humans and livestock. Considering the health and economic importance of the disease, an understanding of the epidemiology of Fasciolosis is highly crucial. This study aimed to evaluate the prevalence and severity of Fasciola infection in animals from different geographical regions of Iran during 2009-10. In this descriptive study, 11100 livers taken from slaughtered sheep and cattle were carefully examined for Fasciola parasites at the six industrial slaughterhouses of East Azerbaijan, Khorasan-Razavi, Khuzestan, Fars, Mazandaran and Markazi provinces. All Fasciola parasites isolated from the livers of infected animals were transferred to the laboratory, and then the parasite species were identified and counted. Finally, the frequency distribution and the severity of infection were analyzed. In this study, 1.10% of the total sheep and cattle slaughtered in six industrial slaughterhouses were found positive for Fasciolosis. The severity of Fasciola in sheep and cattle livers was 7.77 +/- 0.42 and 15.24 +/- 1.78, respectively. Khorasan Razavi and Fars provinces had the highest [14.54 +/- 3.16] and lowest [7.75 +/- 0.79] severity of infection, respectively. Rresults of the study show a reduction in the prevalence and severity of Fasciolosis in sheep and cattle. But considering the importance of the disease and its endemicity, the preventive measures should be taken against the animal and human Fasciolosis in Iran

Molecular identification and differentiation of fasciola isolates using PCR-RFLP method based on internal transcribed spacer [ITS1, 5.8S rDNA, ITS2]

In this study, we used both ITS1 and ITS2 for molecular identification of Fasciola species. The region between 18S and 28S of ribosomal DNA was used in PCR-RFLP method for molecular identification of Fasciola species. Ninety tematodes of Fasciola were collected during abattoir inspection from livers of naturally infected sheep and cattle from Khorasan, East Azerbaijan, and Fars provinces in Iran. After DNA extraction, PCR was performed to amplify region ITS1, 5.8S rDNA, ITS2. To select a suitable restriction enzyme, we sequenced and analyzed the PCR products of F. hepatica and F. gigantica samples from sheep and cattle. Tsp509I fast digest restriction enzyme was selected for RFLP method that caused the separation specifically of Fasciola species. The fragment approximately 1000bp in all of the Fasciola samples was amplified and then digested with the Tsp409I restriction endonuclease. Seventy F. hepatica and 20 F. gigantica were identified of total 90 Fasciola isolates. The new PCR-RFLP assay using Tsp509I restriction enzyme provides a simple, practical, fast, low cost, and reliable method for identification and differentiation of Fasciola practical, fast, low cost, and reliable method for identification and differentiation of Fasciola isolates

Assessment of piwil2 gene expression pattern upon germ cell development from mouse embryonic stem cell

In order to improve culture conditions for optimal spermatogenesis, quantitative assessment of the male germ cell gene expression profile upon spontaneous ES cell differentiation is necessary. In this study, the quantitative expression profile of Piwil2, germ-line specific marker, during the early stage of embryoid body [EB] formation and differentiation [0-3-day-old EB] was studied. CCE mouse embryonic stem cells [ESCs] were cultured in DMED containing 20% fetal bovine serum [FBS] for 1, 2 and 3 days. The total RNA was isolated from ESCs, 1-3-day-old EBs, and adult testis as positive control. cDNA was prepared and quantitative real-time PCR was done for Oct-4 to study the pluripotency of this cell line. Also, the molecular pattern of PiwiI2 expression in developing EB was investigated. Our quantitative results confirmed the pluripotency of CCE mouse ESC line and showed that PiwiI2 was expressed in undifferentiated CCE mouse ESC line. Our results also showed that expression of Piwil2 increased significantly during the process of EB formation and differentiation up to 2-day-old EB and decreased non- significantly in 3-day-old EB. Our results suggest that EB provide a cellular environment similar to the early embryonic microenvironment and cause the efficient and progressive germ cell lineage differentiation in this system