Molecular Characterization of Echinococcus granulosus Sensu Lato from Livestock in North Khorasan Province, Iran.

BACKGROUND: Echinococcus granulosus is one the most important zoonotic disease which is endemic in worldwide. Molecular method has allowed discrimination of different genotypes (G1-G10), providing new approach in development of prevention and control program of hydatid cyst. This study was conducted to identify the genotypes of E. granulosus from domestic animals in nine districts of North Khorasan Province using the mitochondrial cox1 gene sequence. METHODS: Overall, 122 hydatid cyst were collected during 2016-2017 from sheep (n=43) and cattle (n=79). DNA was extracted from protoscoleces and germinal layers and amplified by PCR. Phylogenetic analysis was also performed by analyzing the complete nucleotide sequences of mitochondrial cytochrome C oxidase subunit 1 (cox1) of E. granulosus genotypes from various locations. RESULTS: Sequencing of the amplified products revealed the presence of G1 as dominant genotype, G3 and Echinococcus canadenesis in one isolate each. Altogether, 9 haplotypes were detected based on cox1 gene. Haplotype 3 was the common variant that found in 58 including 42 cattle and 16 sheep. CONCLUSION: This study provided knowledge on the identity of E. granulosus cysts collected from sheep and cattle in North Khorasan Province. Furthermore, these results showed the potentials of sheep as a main source of infection to humans, contributing the transmission and maintain of hydatid cyst in this region.

Molecular characterization of H9N2 avian influenza viruses isolated from vaccinated broiler chickens in northeast Iran.

Avian influenza is a highly contagious respiratory disease of poultry caused by influenza A viruses, family Orthomyxoviridae. H9N2 avian influenza outbreaks are a major problem of the poultry industry in Iran. To determine the genetic differences between field viruses and the vaccine strain, the genomes of four strains isolated in 2011 from vaccinated broiler flocks with a history of respiratory illness were sequenced. Genetic and serological comparisons were made. Sequence analysis of the hemagglutinin (HA) and neuraminidase (NA) genes indicated that the isolated strains shared nucleotide homologies of 91.6-93.9 and 90.2-91.7% with the vaccine strain, respectively. Phylogenetic analyses of HA and NA genes showed that all strains isolated in this study fell into the same group and belonged to the influenza A virus (A)/quail/Hong Kong/G1/97 H9N2 sublineage. Several amino acids have changed at the antigenic sites in HA in the field viruses. Extra potential glycosylation sites were observed in the HA and NA proteins expressed by the current isolates relative to those in the vaccine strain. The deduced amino acid sequence at the cleavage site of HA in recent isolates is the KSSR/GLF motif, whereas it is RSSR/GLF in the vaccine strain. A serological analysis revealed that the currently circulating strains are antigenically distinct from the vaccine strain. These results suggest that the commercial vaccine is insufficiently genetically and antigenically similar to the viruses currently circulating in the region. These findings confirm that it is important to monitor the genetic and antigenic variations in H9N2 influenza viruses when selecting a vaccine strain.

Molecular identification of macroscopic and microscopic cysts of sarcocystis in sheep in north khorasan province, iran.

Sarcocystis is an obligatory intracellular protozoan parasite which can infect humans and animals. Sheep are intermediate hosts of four Sarcocystis species: Sarcocystis tenella, Sarcocystis gigantea, Sarcocystis arieticanis, and Sarcocystis medusiformis The purpose of this study was to perform a molecular identification of the macroscopic and microscopic cysts of Sarcocystis in sheep. In this investigation, the macroscopic and microscopic cysts of Sarcocystis were assessed in slaughtered sheep. The digestion method was used for bradyzoites observation in heart, liver, diaphragm and muscle samples. PCR analysis was conducted on macroscopic and microscopic cysts and also all other samples. Sequencing was performed for ten PCR products. Genotypes were identified by BLAST search and homology analysis. Macrocysts were seen in two muscle tissues. Digestion method and PCR analysis revealed positive results in all samples taken from heart, liver, diaphragm, and muscle. Genotyping of ten tissue samples proved that the genotype of macroscopic belonged to Sarcocystis gigantea and microscopic cysts to Sarcocystis tenella. Microscopic cysts are more prevalent than macroscopic cysts and they can cause enormous economic losses.

First Molecular Identification of Sarcocystis ovicanis (Protozoa, Apicomplexa) in the Brain of Sheep in Iran.

BACKGROUND: The objective of the present study was to survey the presence of Sarcocystis in sheep's brain in North Khorasan Province. METHODS: In general, 80 samples of sheep's brain were collected from slaughtered sheep in slaughterhouses of North Khorasan Province. Tissue digestion method was used for observing bradyzoites in tissues. Histopathological processing tracing Sarcocystis and ensuing structural change in the brain tissue were conducted. PCR analysis was conducted on all the brain samples. Sequencing was done for one PCR product. Genotype was identified by Blast search and homology analysis. RESULT: Sarcocystis spp. was found in one of the brain samples (1.25%) using tissue digestion method. The presence of bradyzoite was also confirmed in the prepared histopathological sections. PCR analysis was positive in one of samples. Genotyping of one sample proved that Sarcocystis species was Sarcocystis ovicanis and the nucleotide sequence of this parasite was deposited in the GenBank database under accession number No.KF489431. CONCLUSION: Sarcocystis ovicanis can involve brain tissue of sheep and consequently causes clinical symptoms.