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Background: Rotavirus (RV) is one of the most important causes of diarrhea in the calf and human neonates. Rotaviruses are divided into nine different serogroups, of which group A is more important compared to other groups. Objective: This study was performed because of the lack of information about the importance and prevalence of bovine rotaviruses B (RVB) and C (RVC) and human genotypes of rotavirus A (RVA) in the bovine population in Iran. Phylogenetic analysis of VP6 of bovine RVA was the second aim of the present study. Materials and Methods: A total of 581 stool specimens were collected from diarrheic calves of 14 provinces and were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and 485 of them were investigated by PAGE electrophoresis to determine the frequency of three rotaviruses A (RVA), B (RVB), and C (RVC). The presence of human G and P genotypes in Iranian bovine population was also evaluated using semi-nested multiplex RT-PCR. Results: RVA was detected by RT-PCR (VP6 gene detection) in 16.2% (94/581) and by PAGE in 22.16% (108/485) and no positive cases of RVB and RVC were confirmed by polyacrylamide gel electrophoresis (PAGE). This study showed that non-A RV groups (B and C) have little role in calf diarrhea in Iran. The results of the phylogenetic study of VP6 sequences of rotaviruses A identified in this study showed that they all belonged to genotype I2 and were classified into three different branches. Specimen isolated in Zanjan showed the highest difference (maximum identity of 94%) with other sequences and clustered along with the Japanese strain, R22. Human G and P genotypes were not found in the studied samples. Conclusion: The results showed that non-A rotaviruses and human genotypes of RVA are of little importance in calf rotavirus diarrhea in Iran. Also, there is the first phylogenetic study of rotavirus A VP6 protein in Iran.

Ebola virus (EBOV) glycoprotein (GP) is a class I fusion protein whose maturation is dependent on furin-mediated processing. EBOV-GP is heavily glycosylated, with glycans constituting ~50% of its molecular mass. Compared with 15 N-linked glycosylation sites, EBOV-GP1 has ~80 potential O-linked glycosylation sites in the mucin-like domain (MLD), suggesting that O-linked glycans are dominated. The membrane-associated RING-CH (MARCH) family consists of 11 members that are RING-finger ubiquitin E3 ligases. Recently, human MARCH1, MARCH2, and MARCH8 were reported to inhibit HIV-1 replication by targeting its Env. Here, we show that human MARCH8 also inhibits EBOV replication by blocking GP incorporation into virions via downregulating its cell surface expression. To understand how the downregulation occurs, we investigated EBOV-GP subcellular localization, processing, glycosylation, and intracellular trafficking in the presence of human MARCH8. We find that MARCH8 interacts with GP and retains GP in the Golgi. MARCH8 also interacts with the homoB domain of furin that blocks its convertase activity. In consequence, MARCH8 blocks GP processing in an MLD-independent manner. Consistently, MARCH8 also blocks the O-linked, but not the N-linked glycosylation of GP. Importantly, in the presence of MARCH8, the shedding of GP1 but not the secreted GP (sGP) is blocked, suggesting that MARCH8 targets the GP1 C-terminal region. The MARCH8 activity is extended to its orthologs from Bos taurus and mice, and its paralogs MARCH1 and MARCH2. In addition, MARCH8 inhibits the processing of two other class I fusion proteins, including HIV-1 Env and IAV HA, and it triggers the degradation of the class III fusion protein VSV-G. We conclude that MARCH8 exerts a very broad and conserved antiviral activity by inhibiting the maturation of class I fusion proteins, which blocks their secretion to the cell surface and incorporation into virions. It should also target class III fusion proteins by triggering their degradation.

Avian influenza (AI) A subtype H9N2 virus belongs to Orthomyxoviridae family and causes low-pathogenic disease AI. The use of gamma-irradiated viral antigens has been developed in the production of effective vaccines. In this research, LPAIV H9N2 strain, A/Chicken/IRN/Ghazvin/2001, was multiplied on SPF eggs and irradiated by a Nordian gamma cell instrument. Irradiated and non-irradiated AI virus (AIV) samples were titrated by EID50 method and hemagglutinin (HA) antigen was analyzed by HA test as the WHO pattern method. Infectivity of irradiated virus was determined by egg inoculation method during four blind cultures. The results showed that after increasing the dose of gamma radiation, virus titer gradually decreased. D10 value and optimum dose for complete virus inactivation were calculated by dose/response curve, 3.36 and 29.52 kGy, respectively. In addition, HA antigenicity of gamma-irradiated virus samples from 0 to 30 kGy was not changed. The results of safety test for gamma-irradiated AIV samples showed complete inactivation with gamma ray doses 30 and 35 kGy, without any multiplication on eggs after four blind cultures. According to the results of HA antigen assay and safety test, the gamma-irradiated and complete inactivated AIV subtype H9N2 is a good candidate as an inactivated immunogenic agent for poultry vaccination.

Avian infectious bronchitis (IB) is an acute and highly contagious viral disease causing severe economic losses in the poultry industry. The 793/B IB virus is an important infectious bronchitis virus (IBV) genotype currently circulating in several countries, including Iran. One hundred confirmed IBV samples (between 2014 and 2015; from 15 provinces in Iran) were selected for genotyping based on S1 sequencing. After phylogenetic analysis, it was found that 30% of the IBV isolates belonged to the 793/B genotype. Results showed that the Iranian 793/B-like IBV isolates could be divided in to three clusters: 4/91-like (50%), 1/96-like (40%), and IB88-like (10%). The sequence similarity between Iranian 793/B-like IBV isolates is 87.69%-100%. The highest identity is between the 4/91 and IB88 clusters (96.38%), and the lowest similarity is between the 1/96 and IB88 clusters (87.62%). This study provides a comprehensive analysis of 793/B-type IBV in Iran and characterization of IBV molecular epidemiology in the country.

Panleukopenia is a viral enteritis of high mortality rate in Feline population. It is caused by a small single-stranded DNA virus, belonging to the Family Parvoviridae, called Feline Panleukopenia Virus (FPV). Parvoviruses are capable of adapting to new hosts and can simply mutate like RNA viruses, resulting in the emergence of different strains with antigenic changes. Unlike the original Canine Parvovirus (CPV) type 2, the antigenic variants (CPV-2a, CPV-2b, CPV-2c) earned the ability to replicate in cat hosts either with clinical signs or without any symptoms. Cats may be able to shed Parvoviruses for a long time and can serve as an important reservoir. Co-infection with FPV and CPV-2 variants has been also reported. These characteristics make the cat an important host for Parvoviruses. Since no molecular study has reported on Parvoviruses infected cats in Iran, a decision was made to study these strains of Parvovirus. Twenty-eight fecal samples were collected from kittens under 6 months of age, both male and female and different breeds which had leukemia. PCR was performed on each sample and 26 samples tested positive for Parvoviruses. Randomly, the PCR products of eight samples were sequenced and their phylogenetic tree was generated. Analysis of the results revealed that all eight samples are FPV and it appears that FPV is more clonal than the CPV-2 variants in Iran. Amino acid residue 232 from the VP2 gene of FPVs has been changed from isoleucine to valine, which has only been reported in CPV-2 and its variants. It seems that some FPVs of Iran are changing to the CPV-2 variants. Hence, it is expected that in the future, the CPV-2 variants will be observed in cats.

Mammalian T7 polymerase-based cytoplasmic expression systems are common tool for molecular studies. The majority of these systems include the internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV). To carry out a cap-independent translation process, this type of IRES might require the expression of an extensive array of host factors, what is a disadvantage. Other IRESes might be less dependent on the host cell factors, but their biology is characterized to a lesser degree. Here, we compare the translational efficiencies of bovine viral diarrhea virus (BVDV) IRES with that of ECMV. Both IRESes were tested in reporter vectors containing the T7 promoter, an IRES of choice and the coding sequence of the enhanced green fluorescent protein (EGFP). To provide for the expression of T7 RNA polymerase, the corresponding gene was isolated from Escherichia coli and inserted into pCDNA3.1-Hygro(+). After co-transfection of the T7 RNA polymerase encoding vector with either of the two IRES-containing reporter vectors into T7 baby hamster kidney (T7-BHK), human embryonic kidney (HEK) 293T, chinese hamster ovary (CHO) and HeLa cells, the translational efficiency of the reporter construct was studied by fluorescence microscopy and flow cytometry. In T7-BHK, HEK 293T and HeLa cells the translational efficiency of BVDV IRES was two to three times higher than that of EMCV IRES. In CHO cells, BVDV IRES and EMCV IRES were equally efficient. An analysis of the secondary structure of respective mRNAs showed that their ΔG values were–544.00 and–469.40 kcal/mol for EMCV IRES and BVDV IRES harboring molecules, respectively. As EMCV IRES-containing mRNA is more stable, it is evident that other, still unidentified factors should be held responsible for the enhanced translational efficiency of BDVD IRES. Taken together, our results indicate the potential of BVDV IRES as a replacement for EMCV IRES, which is now commonly used for T7 polymerase driven cytoplasmic expression of genes of interest or virus cDNA rescue experiments.

Purpose. Bovine leukemia virus (BLV) infects cattle worldwide, imposing an economic impact on the dairy cattle industry. The purpose of this study was to evaluate the molecular epidemiology of BLV in Iran. Methodology. Blood samples taken from 280 cows aged over 2 years old from 13 provinces of Iran were used for leukocyte count and blocking ELISA. Genomic DNA was extracted from the peripheral blood leukocytes of BLV-infected samples and fetal lamb kidney cells to perform PCR of partial env, rex and tax genes and long-terminal-repeat region. The PCR products were sequenced, the phylogenetic tree of each gene was constructed, and nucleotide and amino acid sequence pair distances were calculated. Results. The frequency of BLV infection was 32.8 % among animals and was 80 % among provinces. In BLV seropositive animals, the rate of persistent lymphocytosis was 36.9 %. The constructed phylogenetic trees showed the presence of two BLV genotypes (1 and 4) in Iranian strains. As previous studies, our results showed that the env gene was more variable than previously thought, the Rex protein could withstand more amino acid changes compared to the Tax protein, and no significant differences were observed in average changes of the nucleotide of these genes between clinical stages. Conclusions. Our data indicates an increase in the frequency of this infection in Iran. This is the first study report of the presence of BLV genotype 4 in Iranian farms. These findings may have an important role in the control and prevention of BLV infection in Iran and other countries.

Partial gene sequencing for the bovine coronavirus at the World Genebank is available for many countries, which are distributed unevenly in five continents, but so far, no sequencing of strains has been recorded in Iran. One hundred ninety‐four stool samples from calves with diarrhoea less than one‐month old were collected from five different geographical regions of country in order to detect coronavirus and characterize it if coronavirus was found. Samples were screened for the presence of BCoV by using a commercially available ELISA kit. Furthermore, RT‐PCR was carried out on positive samples for confirmation of the presence of N and S specific genes. Sequencing and phylogenetic analysis was carried out following RT‐PCR tests. 7.2% of samples, were positive for BCoV and all stool samples from the South‐West, Northeast and West regions of Iran were negative. The results showed that all the strains of coronavirus identified in Iran were completely in independent clusters and that they did not stand in the same cluster as any of the strains identified in other parts of the world. The strains from Iran were quite different from strains in other parts of the world but from the point of similarity these viruses showed some similarities to the European strains, such as those found in France, Croatia, Denmark and Sweden., This is the first report of sequencing and phylogeneticic analysis of bovine coronavirus strains isolated from neonatal calf diarrhea in Iran. Sequencing and phylogenetic analysis of BCoV showed that coronaviruses in Iran were quite different from known Bovine coronaviruses (BCoV) in other parts of the world. The phylogenetic tree of gene N indicated that all the viruses identified in Iran were completely in independent clusters and that they did not stand in the same cluster with any of the coronaviruses identified in other parts of the world. Regarding the gene S phylogenetic tree, the coronaviruses were divided into four general groups, with all strains from Iran being grouped together which was different from BCoV isolated in other parts of the world.

The H9N2 avian influenza A viruses (AIV) have been recorded in Eurasia for several years. In this study, the prevalence of the circulated H9 subtype in the poultry population in middle and south of Iraq provinces was studied during a period from September 2014 to June 2015. Samples were col-lected from one hundred broiler flocks with respiratory signs from seven provinces. The detection and identification of virus were carried out by using highly sensitive method, Taqman Real-time Poly-merase Chain Reaction, which has been increasingly used for detecting avian pathogens in recent years. The prevalence of H9 subtype in 16% of the infected flocks was reported, and the results re-vealed that there was a significant difference (P

Background Breast cancer is one of the most common cancers in the world particularly among Iranian women. Bovine leukemia virus (BLV) is an enzootic, exogenous, and oncogenic retrovirus that causes B-cell leukosis in 1–5% of infected cattle. The current study aimed at evaluating the correlation between BLV infection and breast cancer in an Iranian population. Materials and techniques A total of 400 samples including 200 breast cancer-suspected tissue samples and 200 blood samples of women without breast cancer, were collected from July 2017 to October 2018 from women referred to two general hospitals in Qom Province, Iran. The nested PCR technique was performed to determine the presence of tax and gag gene of BLV in the collected samples. Results Out of 200 breast cancer-suspected tissue samples, 172 samples were malignant in terms of pathology. Other samples were reported as non-malignant and non-tumor. Based on nested PCR technique, tax and gag genes of BLV were detected in 30% and 8% of breast cancer-suspected tissue samples, respectively. The frequency of BLV in blood samples collected from women without breast cancer was 16.5% (33/200). Conclusion It seems that human breast cancer and BLV infection in cattle could be associated using nested PCR technique.

Infectious bronchitis virus (IBV) is one of the most critical pathogens in the poultry industry, causing serious economic losses in all countries including Iraq. IBV has many genotypes that do not confer any cross-protection. This virus has been genotyped by sequence analysis of the S1 glycoprotein gene. A total of 100 tracheal and kidney tissue specimens from different commercial broiler flocks in the middle and south of Iraq were collected from September 2013 to September 2014. Thirty-two IBV-positive samples were selected from among the total and were further characterized by nested PCR. Phylogenetic analysis revealed that isolates belong to four groups (group I, variant 2 [IS/1494-like]; group II, 793/B-like; group III, QX-like; group IV, DY12-2-like). Sequence analysis revealed nucleotide sequence identities within groups I, II, and III of 99.68 %-100 %, 99.36 %-100 %, and 96.42 %-100 %, respectively. Group I (variant 2) was the dominant IBV genotype. One Chinese-like recombinant virus (DY12-2-like) that had not been reported in the Middle East was detected. In addition, the presence of QX on broiler chicken farms in the area studied was confirmed. This is the first comprehensive study on the genotyping of IBV in Iraq with useful information regarding the molecular epidemiology of IBV. The phylogenetic relationship of the strains with respect to different time sequences and geographical regions displayed complexity and diversity. Further studies are needed and should include the isolation and full-length molecular characterization of IBV in this region.

Foot-and-mouth disease (FMD) is an extremely contagious viral disease of cloven-hoofed animals that can lead to huge economic losses in the livestock production. No antiviral therapies are available for treating FMD virus (FMDV) infections in animals. The antiviral effects of magnesium oxide nanoparticles (MgO NPs) on the FMDV were investigated in cell culture. The viability of the cells after MgO NP treatment was determined using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The direct effects of MgO NPs on the FMDV in extracellular (virucidal assay) and also different stages of virus replication (antiviral assay) were evaluated by plaque reduction assay. The results showed that MgO NPs were safe at concentrations up to 250 µg/ml in the Razi Bovine kidney cell line. The treatments with NPs indicated that the MgO NPs exerted in vitro virucidal and antiviral activities. Plaque reduction assay revealed that MgO NPs can inhibit FMDV by more than 90% at the early stages of infection such as attachment and penetration but not after penetration. The results of this study suggested that NPs might be applied locally as an antiviral agent in early stages of infection in susceptible animals.

Determination and distribution of the G and P genotypes of group A bovine rotavirus was investigated on 386 fecal samples collected from calves with diarrhea using a semi-nested RT-PCR typing assay. Samples were collected from 11 provinces of Iran during 2010–2012. The provinces divided into 5 different groups based on geographical distance and climates. One hundred and nine samples (28.2%) were confirmed positive for rotavirus group A using ELISA. 75 positive samples were selected randomly and subjected to typing assay. G10 (50.6%) and P[11] (64%) were detected more than G6 (21.3%) and P[5] (9.3%). No any G8 and P[1] were observed. Of the 75 samples analyzed by RT-PCR in each geographical areas named as the south of Alborz mountain ranges area, the north-east area, the central area, the north-west area and the south-east area, number of samples with G10 genotype were 19, 9, 9, 0 and 1; G6 were 8, 0, 3, 5 and 0; P[11] were 25, 7, 11, 5 and 0 and finally P[5] were 5, 0, 2, 0 and 0 in each area, respectively. The most common VP7/VP4 combinations were G10P[11] (40%), G6P[11] (12%), G6P[5] (5.3%) and G10P[5] (2.6%). Phylogenetic analysis of one strain showed high identity with strain B223. Since the identification of G and P genotypes and their diversity is fundamental to development and use of effective vaccines, we determined the most prevalence G and P genotypes of bovine rotavirus group A (BRVA) in a broad area of Iran.

The most important causal agent of border disease (BD), border disease virus (BDV), is a Pestivirus in the family Flaviviridae. In attention to the lack of molecular documented data on the prevalence of BDV in Iran, especially in East Azerbaijan Province, northwest of Iran, this research was designed to determine the prevalence of BDV in suspected ewes that have an antecedent of reproductive failure. Eighty-eight suspected ewes in the breed of Ghezel, Mogan, and mix (Ghezel × Mogan) in five various districts of East Azerbaijan Province (Azershahr, Lyqvan, Osku, Sofian, and Varzqan) were selected. Razi bovine kidney (RBK) cell lines were grown, and the buffy coat layer was separated and inoculated to the monolayer cell culture. All cell cultures were inspected for cytopathic effects after 5–7 days. BDV genomes were being checked using nested reverse transcription polymerase chain reaction method. Ten samples (10/88, 11.36 %) were positive for the genome of BDV. The prevalence of BDV infection was found to vary between different districts. The highest percentage of infected sheep was in animals at the 3–4.5-year-old range (15.38 %). There was no statistically significant difference in the prevalence among different age categories. The highest BDV infection rate was observed in full blood ewes in Mogan and Ghezel. Cytopathic effects of the virus in the culture on RBK cell line were not observed at all. This study proved that the infection of BDV is present in sheep in the East Azerbaijan, northwest of Iran, and cause some reproductive failure in the flocks. The survey is the first molecular evidence of the occurrence of the disease in the country.

H9N2 avian influenza viruses (AIVs) have been recorded in Eurasian for several years. Since 2004-2005, the disease has become endemic in Iraq, causing serious economic losses in the poultry industry. The hemagglutinin (HA) and neuraminidase (NA), two out of eight protein-coding genes, play an important role during the early stage of infection and hinder virus assembling. Little is known about the genetic information of the H9N2 viruses currently circulating in Iraq; thus, gene sequences of six AIVS of the H9N2 subtype have been detected and analyzed in the period of 2014-2015 from different outbreaks of broiler flocks in five provinces situated in the middle and southern parts of Iraq. Genetic comparison of the partial sequences of HA gene indicated that all Iraqi viruses are related to each other and could be divided into two subgroups. Viruses of the first and the second subgroups demonstrated a high similar identity with Pakistani and Iranian viruses, respectively. The nucleotide sequences of the NA protein of the all studied Iraqi viruses were very similar (95.2-100% identity), and shared high nucleotide sequence identity with Iranian, Pakistani, and Lebanese strains. All six recent viruses possessed histidine, alanine, and leucine at positions 183, 190, and 226, respectively, which are the key residues in receptor-binding sites. The Iraqi viruses were closely related to viruses of G1-like lineage isolated from poultry flocks of Iran and Pakistan, suggesting that possible epidemiological links could be derived from a common origin. Further investigations are required and should include the viral isolation and full-length molecular characterization of H9N2 AIVs in this area.

The aim of this study was to investigate the frequency of the Newcastle disease virus (NDV) infection and its virulence in exotic cage birds over a limited area and time period. A set of 335 samples was collected from 24 different species of exotic unvaccinated cage birds kept in the zoological gardens and bird markets of the Tehran province of Iran during 1.5 years. Except for three pigeons, all of the sampled birds were healthy with no clinical signs of Newcastle disease. NDV was detected in three sick pigeons by haemagglutination assay (HA), haemagglutination inhibition (HI) and reverse transcription-polymerase chain reaction (RT-PCR) tests while two of them were identified as virulent types by RT-PCR. Although the remaining samples were negative by Newcastle-disease-specific HA and HI tests, 35 of them (10%) were identified as positive and 25 (72%) were determined as the velogenic type by RT-PCR test. Five PCR products were sequenced and all were confirmed as NDV but sequences were different from each other and from other sequences from Iran. In total, 14 species (58%) were infected and 10 species were uninfected with the velogenic type without showing any signs. Pigeons are very sensitive to NDV infection and play an important role in its epidemiology. In this study, the PCR test was found to be a more sensitive and powerful method than the HA and HI tests for detection of NDV reservoirs and carrier status in exotic birds. Also, the frequency of infection with the virulent type showed that the exotic birds should probably be considered one of the main causes of recurrent annual epidemics of Newcastle disease in endemic regions.

Newcastle disease (ND), caused by avian paramyxovirus type 1, is a highly contagious and devastating viral disease of poultry of worldwide distribution with an enormous economic impact. In this study, the sequences of matrix protein (M) gene of three ND virus strains isolated from outbreak in chickens in poultry farms of Iran in 2011–2012 were determined. Phylogenetic analysis revealed that both strains clustered with the class II viruses, with one phylogenetically close to the genotype VIIb Newcastle disease viruses and the other closer to genotype VIId. This finding is essential for improving the disease control strategies and development of vaccines for ND. The study results thus emphasize the importance of continuous surveillance of this disease and of sharing the information to the global scientific community, which would help to fill the epidemiological gaps in the regions and to validate the robustness of diagnostic screening.

One of the important issues of food hygiene is Salmonella contamination of eggs that may cause food-borne infection and disease in humans. The aim of this study was to investigate Salmonella contamination of traditionally produced poultry eggs in Tehran. Contamination of eggs to other Enterobacteriaceae was also investigated. For this purpose, 200 eggs including 131 egg samples from chickens and 69 from quails, geese, and ducks (23 samples from each species) were investigated. After conventional isolation procedures, Salmonella contamination was detected in five chicken eggs. Serological tests revealed that all of the isolated genera belonged to D serogroup and multiplex PCR showed that all strains carried spv and sefA genes and also random sequence (specific for the genus Salmonella); therefore, all strains confirmed as Salmonella Enteritidis. Antimicrobial susceptibility test revealed sensitivity of all Salmonella isolates to florfenicol, ceftriaxone, trimethoprim–sulfamethoxazole, chloramfenicol, and oxytetracycline. In addition, except the quail eggs, 29 enteric bacteria were isolated from eggs including 21 Enterobacter spp., 4 Klebsiella spp., 3 Escherichia coli, and 1 Proteus spp. This study indicated that traditionally produced poultry eggs were highly contaminated by Enterobacteriaceae. Moreover, the chicken eggs were contaminated by Salmonella Enteritidis; therefore, non-commercial chicken eggs can be considered as an important threat for public health.

Bovine viral diarrhea virus (BVDV) is an economically important animal pathogen affecting cattle. Despite the use of vaccination, test and slaughter practices, BVD remains a serious problem of cattle breeding. This study was conducted in order to develop a cell line that expresses some of BVDV sub-replicons. BVDV-NADL NS3 and 5’UTR were cloned in pWPI-linker B lentiviral plasmid at the upstream of EGFP gene. Consequently, lentiviral vectors containing BVDV-NS3 and BVDV-5’UTR were produced by using the second-generation lentiviral packaging system. By these lentivectors, MDBK cells expressing BVDV-5’UTR and BVDV-NS3 partial fragments were prepared. The efficiency of the infection was evaluated by fluorescence microscopy, western blotting, and RT-PCR. The results indicated that the development of MDBK cell line expressing these transgenes provides a very sensitive antiviral drug screening system for anti-bovine viral diarrhea virus (BVDV) therapies.

Analyses for alanine aminotransferase, aspartate aminotransferase (AST), alkaline phosphatase (ALP) activities, and iron concentration were carried out in infected and uninfected cattle in order to determine the degree of hepatic damage and blood iron status caused by Theileria annulata and Babesia bigemina. Blood samples from 30 infected and 20 uninfected cattle with T. annulata and also from 40 infected and 20 uninfected cattle with B. bigemina were taken. The biochemical analyses revealed increased serum AST and ALP activities in infected cattle with T. annulata and B. bigemina compared with uninfected cattle (P < 0.05). Significant decreases in iron concentration were observed in cattle infected with T. annulata and B. bigemina when compared with uninfected cattle (P < 0.05) although the level of serum iron concentration in infected cattle was within the normal reference values for cattle. It was concluded that iron deficiency anemia is not an important factor in T. annulata- and even in B. bigemina-infected cattle with severe intravascular hemolysis. The increased activities of serum enzymes indicated the hepatic injuries associated with infection with T. annulata and B. bigemina.

High mortalities in 17 canary flocks from different regions of Tehran, Iran, were reported. Necropsy and histopathologic examination revealed necrotic hepatitis and overall congestive septicaemia in carcasses. Salmonella enterica was isolated from 34 examined samples, two samples from each flock, including visceral organs of carcasses and droppings of live diseased birds. All isolates were typed as Salmonella enterica serovar Typhimurium by conventional serotyping. Antibiotic resistance profiling using 33 antibiotics and random amplification of polymorphic DNA differentiation by three primers were performed and showed an identical clonal relationship between these isolates and S. Typhimurium isolated from a sample of feedstuffs. Changing the feed ingredients along with antibiotic therapy via the drinking water by enrofloxacin solution controlled the outbreaks, and mortalities ceased. The zoonotic nature of S. Typhimurium and close contact of bird owners with pet birds in the home environment made the case significant in relation to public health.

Genomic and phenotypic evaluation of Salmonella typhimurium and Salmonella enteritidis by two simple, fast, and applicable methods of random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) and antibiotic susceptibility for employment in epidemiologic surveillance screening systems in Iran was the objective in this study. We selected 60 (30 S. typhimurium and 30 S. enteritidis) isolates from different animal sources and different times and places among 1975–2006 in Iran. Serotyping with traditional method and reliable antisera was done and then confirmed with multiplex PCR (with four and three pair of primers, respectively). All of S. typhimurium and S. enteritidis isolates have virulence genes of invA and spv, respectively. Then, from nine primers, two primers that have enough discriminatory power were selected for RAPD analysis. We set up the quality and quantity of DNA template, primers, MgCl2, Taq DNA polymerase, and deoxyribonucleotide triphosphates concentration for RAPD analysis. We also examined the strains with disk diffusion standard antibiotic susceptibility test. With the application of primer P1254; we saw four and seven and primer 23L, six and three profiles in RAPD analysis and 13 and six profiles of R-type in susceptibility test of S. typhimurium and S. enteritidis, respectively. Combination of two methods differentiated these 30 strains to 20 and 16 different profiles, respectively. The finding of this study indicated that clonality of S. enteritidis is more than S. typhimurium in Iran and RAPD-PCR in combination with antibiotic susceptibility test were fast, easy, relatively reliable, and discriminatory molecular and phenotypic methods in the differentiation of S. typhimurium and S. enteritidis for epidemiologic purposes in Iran.

Infectious bronchitis (IB) is a viral avian disease with economic importance in the world, including Iran. S1 gene sequencing has been used for molecular epidemiological studies and genotypic characterization of infectious bronchitis virus (IBV). A total of 118 IBV isolates were obtained from tissue samples from chickens with clinically suspected IB from Iranian broiler farms (eight provinces, 200 samples). The isolates were confirmed by real-time polymerase chain reaction (PCR) and characterized by sequencing the spike glycoprotein gene. The isolates formed six distinct phylogenetic groups (IS/1494/06 [Var2] like, 4/91-like, IS/720-like, QX-like, IR-1 and Mass-like) that were related to variants isolated in the region. The most frequently detected viruses were of the Var2-like (IS/1494/06-like) genotype, with an overall prevalence of 34 %. Twenty-one percent of the isolates formed a cluster together with the 4/91 IBV type, 10 % were of the QX genotype, and 8 % were of the IS/720 genotype. In addition, 4 % and 3 % of the isolates belonged to the Massachusetts and IR-1 genotype, respectively. For the first time, we have isolated and characterized IBV variants from broiler farms in different provinces of Iran. This study demonstrates a constant evolution of IBV in Iran, demonstrating the need for continuous monitoring and development of new vaccines based on indigenous viruses.

Two experiments were conducted to investigate the effects of fortification of the pre-starter diet with glutamine in post-hatch broiler chicks. In the first experiment, male chicks were used to study the impact of glutamine supplementation at three levels (0%, 1% and 2%) on performance, apparent faecal digestibility (AFD) and ileal digestibility (AID) of amino acids (AAs), as well as intestinal morphometric parameters from 0 to 14 days old. The best performance was obtained at 1% supplementation. Average daily feed intake and feed conversion ratio (FCR) were affected by treatments. Supplementing diets with glutamine resulted in increase of digestibility values for arginine (Arg), lysine (Lys), isoleucine (Ile) and valine (Val). Glutamine-supplemented diets led to higher villus height (VH) and shallower crypt depth (CD) in the jejunum and ileum. A second experiment was performed to investigate whether the form of glutamine could affect performance and morphometric parameters in newly hatched chickens. A control diet was prepared, based on casein as the sole source of protein-bound AAs. Three diets based on synthetic AAs were used to obtain nutrient values of the control diet, which were supplemented with a synthetic form of glutamine, glutamic acid and a 50 : 50 ratio of glutamine : glutamic acid. Chickens fed the control diet presented higher average daily weight gain and better FCR. None of the AID coefficients of amino acids were influenced by the dietary treatments, except glutamic acid and aspartic acid, which were decreased in chicks fed synthetic AAs. In conclusion, 1% glutamine supplementation improved performance, morphometric parameters and AID and AFD values of cationic AAs. The form in which dietary glutamine was provided could affect performance. Keywords: Amino acid, broiler, digestibility, intestinal morphology

Salmonella infection is one of the most important diseases that affect all animal species and is the leading cause of foodborne infections worldwide. One of the challenges in the control and prevention of salmonellosis is the recognition of potential carriers. To assess the potential role of shepherd dogs in the epidemiology of salmonellosis, fecal samples were taken from 38 shepherd dogs in Garmsar Province. Salmonella strains were isolated from 4 dogs (10.5%) related to 3 sheep flocks. All of the strains were confirmed at the genus level using invA virulence gene PCR. Serotyping confirmed all of the strains as Salmonella Reading. To study the clonal relatedness of the isolates, the strains were subjected to RAPD-PCR, and antimicrobial resistance patterns were also determined using 9 antibacterials. The similar RAPD-PCR profiles and resistance patterns strongly suggested that the shepherd dogs in the present study shed a similar clone of S. Reading; therefore, shepherd dogs can be recognized as a common source for this serotype in Garmsar.

Upper respiratory tract diseases (URTD) are common clinical problem in cats worldwide. Feline calicivirus (FCV) and feline herpesvirus type 1 (FHV-1) are the main primary pathogens. Feline immunodeficiency virus (FIV) and Feline leukemia virus (FeLV) are also among the most common infectious diseases of cats which suppress the immunity. Oropharyngeal and conjunctival swabs and blood samples were taken from 16 cats with clinical signs of URTD and 26 clinically healthy cats. PCR and RT-PCR were used to detect FHV/FIV or FCV/FeLV infections, respectively. Feline calicivirus was detected in all cats with URTD and 87.00% and 93.00% of them were positive for FIV and FeLV, respectively. Feline herpesvirus rate of infection was 43.00% in sick cats. In clinically normal cats, prevalence rates of FCV and FHV were about 50.00%, but FIV and FeLV rates (42.00% and 65.00% respectively) were higher compared to other studies. Stomatitis was observed in 50.00% of cats with URTD. The main causative agent of corneal ulcers is FHV-1, but in 50.00% of cats with corneal ulcers, FCV was detected alone. It seems new variants of Caliciviruses are the main causative agents to attack uncommon tissues like cornea, although retroviral infections may be in the background of these various signs. The high retroviral prevalence may be due to existence of large population of stray cats. This is the first molecular study of FeLV and FCV in Iran and seems that FCV and FHV prevalence rates in FIV or FeLV infected cats is more than other non-infected ones.

The purpose of this study was to determine the utility of RAPD-PCR and antibiotic susceptibility tests in differentiation of S. typhimurium isolates in Iran. Thirty isolates of S. typhimurium, selected based on animal source; place and time of the isolation and a reference strain of the bacterium were used in this study. Serotyping of the isolates was performed by reliable antisera and confirmed with a multiplex PCR. Genomic DNAs were extracted and subjected to an optimized RAPD-PCR, using two previously reported arbitrary primers (P1254 and 23L). Bacteria were also examined for resistance against 8 antibiotics, using a standard antibiotic susceptibility test. While the antibiotic susceptibility test resulted in the identification of 13 profiles of R-type among the bacterial isolates, application of primers P1254 and 23L in the RAPD-PCR could discriminate the isolates only in four and six profiles respectively. However, combination of the two methods could differentiate the 30 isolates in 20 different profiles. The results of this study indicate that the discriminatory power of RAPD-PCR for S. typhimurium is low but a combination of this method with antibiotic susceptibility test could be considered an easy and relatively reliable discriminatory approach in differentiation of S. typhimurium for epidemiologic purposes in Iran.