Characterization of Eimeria Species in Commercial Broilers by PCR Based on ITS1 Regions of rDNA

Background: Coccidiosis is an intestinal disease of chickens caused by various species of proto­zoan parasites within the genus Eimeria. Diagnosis and genetic characterization of different spe­cies of Eimeria are central to the prevention, surveillance, and control of coccidiosis. The aim of this study was to detect different chicken Eimeria species from several areas in Khuzestan, south­west Iran.Methods: From February to September 2008, PCR assay as well as parasitological examinations was applied for the identification of field isolates of Eimeria parasites around Ahvaz, center of Khuzestan, southwest Iran. Data were analyzed by the Kappa statistic test.Results: Eimeria maxima, E. necatrix, E. tenella, E. acervulina and E. mitis were detected in this study. The prevalence of Eimeria spp. was 31.5% (126 of 400) and E. tenella was the most preva­lent species in Khuzestan. Based on the Kappa statistical test, a good correlation between the results of PCR and traditional biometrical methods was only observed for E. maxima.Conclusion: The present study is the first on the prevalence of Eimeria species in Khuzestan, based on the molecular findings. We believe that traditional methods are not sufficiently reliable for specific diagnosis of Eimeria species in chickens and PCR based amplification of DNA se­quence of parasite, could resolve this problem

Molecular cloning of S1 glycoprotein gene of infectious bronchitis virus (IBV) serotype 793/B in secretory Pichia pastoris vector

In vitro protein expression is an important method of obtaining large amounts of viral proteins to investigate their biological properties. The S1 glycoprotein of infectious bronchitis virus, due to its effective immune-dominant role is an appropriate candidate for production of recombinant vaccine against infectious bronchitis disease. In this study, the S1 gene fragment of infectious bronchitis virus strain793/B was amplified using reverse transcriptase-polymerase chain reaction (RT-PCR) and purified. Itwas then cloned into pPICZαA a secretory expression vector of Pichia pastoris. The insertion was proved by PCR analysis and isolation of gene from construct by restriction enzymes and finally, it was sequenced. After the expression of S1 gene in P. pastoris expression system, it was found that it could be used in the production of recombinant vaccines against infectious bronchitis disease. Keywords: Infectious bronchitis, S1 glycoprotein, cloning, Pichia pastori

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

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

The economics of biomass production in the United States

Biomass crops (e.g. poplar, willow, switchgrass) could become important feedstocks for power, liquid fuel, and chemical production. This paper presents estimates of the potential production of biomass in the US under a range of assumptions. Estimates of potential biomass crop yields and production costs from the Department of Energy`s (DOE) Oak Ridge National Laboratories (ORNL) are combined with measures of land rents from USDA`s Conservation Reserve Program (CRP), to estimate a competitive supply of biomass wood and grass crops. Estimates are made for one potential biomass use--electric power production--where future costs of electricity production from competing fossil fuels set the demand price. The paper outlines the methodology used and limitations of the analysis

Expression of the G1 epitope of bovine ephemeral fever virus G glycoprotein in eukaryotic cells

The envelope glycoprotein (protein G) of bovine ephemeral fever virus (BEFV) has been identified as a plausible vaccine candidate against the BEF disease. In the present study, G1 epitope of the G gly-coprotein gene was cloned in an eukaryotic expression vector, pcDNA3.1(+), under the control of the human cytomegalovirus (CMV) promoter. The pcDNA3.1-G1 construct was transfected into human embryonic kidney 293 (HEK 293) cell line and the expression efficiency was verified by immunofluorescence staining of transfected cells and Western blot analysis. The results indicated that G1 protein was expressed by the recombinant pcDNA3.1-G1 construct in the transfected cells. The recombinant plasmid constructed in this study can be used as a DNA vaccine to evaluate its potential for immunogenicity and protection against BEF virus in animal models

Transcriptional Landscape of the Prenatal Human Brain

Summary The anatomical and functional architecture of the human brain is largely determined by prenatal transcriptional processes. We describe an anatomically comprehensive atlas of mid-gestational human brain, including de novo reference atlases, in situ hybridization, ultra-high resolution magnetic resonance imaging (MRI) and microarray analysis on highly discrete laser microdissected brain regions. In developing cerebral cortex, transcriptional differences are found between different proliferative and postmitotic layers, wherein laminar signatures reflect cellular composition and developmental processes. Cytoarchitectural differences between human and mouse have molecular correlates, including species differences in gene expression in subplate, although surprisingly we find minimal differences between the inner and human-expanded outer subventricular zones. Both germinal and postmitotic cortical layers exhibit fronto-temporal gradients, with particular enrichment in frontal lobe. Finally, many neurodevelopmental disorder and human evolution-related genes show patterned expression, potentially underlying unique features of human cortical formation. These data provide a rich, freely-accessible resource for understanding human brain development