Sequence analysis of ORF94 in different white spot syndrome virus (WSSV) isolates of Iran

White spot syndrome virus (WSSV) is a pathogen that causes high mortality in shrimp culture in the whole world. Sequence analysis of WSSV has shown similarity of WSSV isolates in different countries with exception of a few variable genomic loci. This study investigated the sequence variation of some Iranian WSSV isolates and previously identified isolates. Samples were collected during target surveillance and were feed, broodstock, post-larvae, artemia, crabs, and wild and cultured shrimp of northern Persian Gulf (Boushehr and Khuzestan provinces). The open reading frame (ORF) 94 sequence of different Iranian WSSV isolates were amplified using specific primers from positive samples. The ORFs 94 sequence of positive samples were sequenced and registered in the Gene Bank and then compared to other WSSV isolates. The number of repeat units in ORF94 showed that WSSV isolates were varied in number. There are SNPs (G and T) in position 48 of RUs that varies in different Boushehr and Khuzestan isolates. Also these sequences were compared to Gene Bank WSSV isolates and showed a high similarity (>90%) to Southeast Asian countries. To our knowledge this is the first report of sequence analysis in Iranian WSSV isolates applications

Isolation and expression of recombinant viral protein (VP2) from Iranian isolates of Infectious Pancreatic Necrosis Virus (IPNV) in Escherichia coli

Infectious Pancreatic Necrosis Virus (IPNV) is a member of the family Birnaviridae that has been linked to high mortalities in salmonids. Bacterial based systems as live vectors for the delivery of heterologous antigens offer a number of advantages as vaccination strategies. VP2 is a structural viral protein of IPNV with immunogenicity effects. In this study IPNV was isolated from diseased fry of rainbow trout Oncorhynchus mykiss (Walbaum) using CHSE-214. Then an expression vector was constructed for expression of viral protein VP2. The designed vector was constructed based upon pET-26b (+) with T7 promoter. A fragment containing the full length of the VP2 gene of Iranian Sp strain was amplified by PCR using genomic RNA of IPNV as template and cloned inpET-26b(+) plasmid. Recombinant structural viral protein VP2 was expressed as a soluble, N-terminal PelB fusion protein and secreted into the periplasmic space of Escherichia coli BL21(DE3) and Rosetta (DE3). The glucose, Isopropyl-β-D-thiogalactopyranoside (IPTG) was used as a chemical inducer for rVP2 production in 37º C. The rVP2 was extracted from the periplasm by osmotic shock treatment. The presence of gene in bacterial system of E. coli was confirmed by gel electrophoresis technique. The constructed vector could efficiently express the rVP2 into the periplasmic space of E. coli. The successful cloning and expression of the structural viral protein gene into E. coli can be used for developing a useful and safe vaccine to control IPNV infection in Iranian fish industry

Phylogenetic relationships of Iranian Infectious Pancreatic Necrosis Virus (IPNV) based on deduced amino acid sequences of genome segment A and B cDNA

Infectious Pancreatic Necrosis Virus (IPNV) is the causal agent of a highly contagious disease that affects many species of fish and shellfish. This virus causes economically important diseases of farmed rainbow trout, Oncorhynchus mykiss, in Iran which is often associated with the transmission of pathogens from European resources. In this study, moribund rainbow trout fry were collected during an outbreak of IPNV in three different fish farms in one northern province (Mazandaran), and two west provinces (Chaharmahal and Bakhtiari, and Kohgiluyeh and Boyer Ahmad) of Iran. We investigated full genome sequence of Iranian IPNV and compared it with previously identified IPNV sequences. The sequences of different structural and non-structural protein genes were compared with other aquatic birnaviruses sequenced to date. Our results showed that the Iranian isolate fall within genogroup 5, serotype A2 strain SP, having 99 % identity with the strain 1146 from Spain. These results suggest that the Iranian isolate may have originated from Europe

Comparison of serum homocysteine levels in patients with coronary artery disease with and without diabetes mellitus

Background: Homocysteine is an amino acid that is produced during the metabolism of the methionine cycle. In previous studies, a causal role of homocysteine in coronary artery disease (CAD) has been investigated; however, the homocysteine level in diabetic and non-diabetic patients with CAD has not been compared. This study aimed to investigate this issue. Materials and Methods: This cross-sectional study was conducted on 200 patients with CAD in Kashan Shahid Beheshti Hospital during 2014-2015. Coronary artery disease was confirmed by angiography. Homocysteine levels, blood urea nitrogen, creatinine, hemoglobin A1C and mean blood pressure were measured. Data were analyzed using t-test with SPSS version 16. Results: In this study, one hundred and eight of patients (54) were men. The mean concentrations of hemoglobin A1C in diabetic and non-diabetic patients were 6.69±1.44 and 5.74±0.92, respectively (P<0.001). Moreover, the mean homocysteine serum levels in diabetic and non-diabetic patients were 19.89±6.86 μmol/L and 24.35±9.93 μmol/L, respectively (P<0.001). Conclusion: Results of the current study showed that the homocysteine serum level in patients with CAD was higher than the normal level and in patients without diabetes was significantly higher than patients with diabetes. Also, in diabetic patients with CAD, the serum creatinine and urine protein levels were higher than those in non-diabetics patients