Comparison of Penton and Hexon Genes of Fowl Adenovirus Serotype 11 in Molecular Detection of IBH in Broilers.

Fowl Adenoviruses (FAdVs) are widely distributed pathogens across the globe. The FAdVs from serotypes FAdV 2, 3, 8a, 8b, 9, and 11 are responsible for inclusion body hepatitis (IBH). Recently, increased mortality and IBH-suspected lesions were observed in 8-10-day-old broiler chickens in West Azerbaijan Province, Iran. In this regard, the present study aimed to compare penton and hexon genes of ADDV11 in the molecular detection of IBH in broiler chickens. In total, 100 liver specimens were collected from 10 suspected farms, and their DNAs were extracted. Two polymerase chain reactions (PCRs) were applied; one targeting the L1 region of the hexon gene and another aiming at the penton gene. Based on the findings, 60% of samples showed positive results in both PCRs and phylogenetic analysis clustered the studied viruses into serotype 11 (species D) FAdV. The detected FAdVs also shared a multitude of homologies with previously published serotype 11 viruses from Iran and those identified in Pakistan, Saudi Arabia, India, China, and Canada. This research not only provides an update on circulating FAdVs in Iran, but also introduces the penton gene as an alternative target for IBH diagnosis. Considering that IBH is a primary disease in Iran with both horizontal and vertical routes of transmission, urgent preventive measures are needed.

Molecular Detection of Avian Infectious Bronchitis Viruses in Live Bird Markets, Gilan Province.

Coronaviruses (AvCoV) which include infectious bronchitis virus (IBV) and other bird coronaviruses belong to the genus gammacoronavirus, subfamily Coronavirinae. One of the most prominent representatives of gammacoronavirus genus is infectious bronchitis virus (IBV) which is a highly contagious viral pathogen of chickens causing considerable economic losses to the poultry industry. IBVs mostly affect the respiratory, urinary, and reproductive tracts leading to a substantial drop in production. Backyard poultry in the villages usually share their food and water with free flight birds which puts them at serious risk of disease transmission. Furthermore, the poor hygienic measurements which are often used in backyard flocks make them a potential reservoir for diseases that can be transferred to commercial poultry flocks. Live bird markets (LBMs) which receive live poultry to be resold or slaughtered and sold onsite play a significant role in spreading infectious diseases among the different bird species. In the present study, a number of 354 cloacal swab samples were collected from different bird species from LBMs of Gilan province. Subsequently, after RNA extraction, reverse transcription-polymerase chain reaction (RT-PCR) technique was carried out using specific primers of S1 gene to detect coronavirus infectious bronchitis virus. Two samples from backyard chickens were reported to be positive to coronavirus which were named Iran/Backyardchicken 96/2017 and Iran/Backyardchicken 94/2017. The results of the phylogenetic analysis demonstrated that these two isolates are placed in QX and IS-1494 strains, respectively. On a final note, the obtained results highlighted the role of live birds offered in LBMs in the epidemiology of IBV and the transmission of the virus to the industrial flock.

Prevalence of Avian Influenza in Live Bird Markets, Bird Gardens, and Zoos in Iran in 2015: A Cross-sectional Study.

Avian Influenza (AI) H9N2 is endemic in Iran; therefore, it is necessary to estimate the disease prevalence among birds in live bird markets (LBMs) and assess the risk spread across the country. Accordingly, this study aimed to estimate the prevalence of AI subtypes in LBMs, bird gardens, and zoos during October and November 2015 in Iran and investigate the associated risk factors. Data related to independent variables for birds and premises were collected using a prepared questionnaire which included items about previously known potential risk factors associated with avian influenza infection. Serological testing was carried out to detect the antibodies against H5, H7, and H9. Regarding H5 and H7, the antigens H5N2 and H7N1 were used in this study. Positive samples on the first test were examined with the second antigens, namely H5N1 and H7N7. Moreover, sera with titers ≥4 (i.e. log2) were considered positive and premises with at least one positive bird were considered as positive units. In total, 87 premises were included in this cross-sectional study. Serum samples were examined utilizing hemagglutination inhibition, and RT-PCR was conducted on swab samples. Regarding the molecular test, the RNA was extracted using the High Pure Viral RNA Kit (Roche, Germany). In addition, real-time RT-PCR was conducted based on the described method. The seroprevalence rates of H9N2 were 83.9% and 31.8% at the premises and bird levels, respectively. Totally, 9.2% of pooled swab samples were positive for H9N2. However, all sera and swab samples were negative for H5 and H7. Hot and humid weather (OR=0.13, 95% CI 0.02 – 0.78) as well as bird-keeping condition (i.e., enclosed area) (OR=0.11, 95% CI 0.012 – 1.02) were protective factors for H9N2. High seroprevalence rate of H9 indicates that the disease is endemic in Iranian LBMs. Active surveillance must be carried out in LBMs, especially in the northern provinces of Iran. In addition, cleanliness and improved hygiene would be useful to prevent the spread of disease in LBMs.

Molecular Detection of Gamma Coronaviruses in Bird Parks of Iran.

Gamma Coronaviruses (GCoVs) are distributed worldwide, affecting a wide range of bird species, the beluga whale, and bottlenose dolphins. Because of the limited proofreading capability in the viral encoded polymerase, they emerge genetically diverse. There has been no molecular surveillance data to describe the epidemiology of GCOVs in avian species. The present study was conducted to detect GCOVs in Tehran birds’ parks, 2015. Cloacal swabs (267 samples) from eight different bird species ((Chickens (Gallus gallus), Pheasant (Phasianus colchicus), Turkey (Meleagris gallopavo), Partridge (Perdix perdix), Quail (Coturnix coturnix), Duck (Anas platyrhynchos), Goose (Anserini),and Guinea fowl (Numididae)) were collected, the viral RNA was extracted, the RT-PCR was performed using QIAGEN one step RT-PCR kit and the primers targeting “3'-UTR” and “Nucleocapsid” genes. The detection rate was approximately 8.99%. GCOVs were detected in the chicken, quail, pheasant, turkey, and the partridge with different prevalence rates. Phylogenetic tree based on partial nucleotide sequences of the N gene clustered the samples into two groups. It is the first report of GCOVs in non-commercial birds in Iran. According to our results, GCOVs are circulating in different avian species, and further studies are needed to isolate these viruses and evaluate their pathogenesis.

Serological Survey of Avian Influenza (H9N2) in Commercial Ostrich Farms in Iran, 2015.

The aim of this study was to determine the seroprevalence of avian influenzaH9N2 subtype in the industrial ostrich farms and its geographical distribution. This cross-sectional study was conducted from January to June 2015. A total of 40 farms were selected from different provinces of Iran, from each of which 11 ostriches (n=440) were sampled. The sera samples were examined using 4 hemagglutination units of H9N2 antigens. A frequency distribution was used to describe the responses to the survey questions. The mean titers between provinces were compared using one-way analysis of variance. According to the results, 21 (47.5%) out of 40 farms and 108 (24.5%) out of 440 ostriches tested positive in the HI-H9N2 test. There were statistically significant differences between the mean titers of samples in different provinces (P<0.001). The current study was conducted on unvaccinated ostriches. The results showed that H9N2 had a high seroprevalence at both farm and bird levels. The findings of this study can be for the further investigation of infection in ostrich farms in order to consider this species in the surveillance programs of the Iranian Veterinary Organization. The detection and isolation of viruses and epidemiological investigation are necessary for the persistent use of H9N2 vaccines in some ostrich farms.

Co-administration of ginseng and ciprofloxacin ameliorates epididymo-orchitis induced alterations in sperm quality and spermatogenic cells apoptosis following infection in rats.

Korean red ginseng (KRG) may be a beneficial adjuvant along with ciprofloxacin to ameliorate devastating effects of epididymo-orchitis (EO) on male fertility. This study intends to assay the effects of KRG and ciprofloxacin on sperm quality and spermatogenic cells apoptosis in EO rats. We divided 54 adult rats into nine groups (n = 6 rats per group): control (CO), sham-operated (SH), EO (E); ciprofloxacin (C), EO-ciprofloxacin (EC), KRG (G), EO-KRG (EG), ciprofloxacin-KRG (CG) and EO-ciprofloxacin-KRG (ECG). We administered ciprofloxacin and KRG 48 hr after the Escherichia coli (E. coli) injection for 10 days. Bilateral orchiectomy was performed after one sperm cycle (14 days) following the last treatment with ciprofloxacin and KRG. Total and progressive motility of E, C and EC groups decreased. However, motility is improved in CG and ECG in comparison with these groups. The E group induced negative changes in the architecture of testes tissue and dramatic increase in apoptosis indices. Interestingly, co-administration of ciprofloxacin and KRG has dramatically improved Miller's and Johnsen's scores and decreased the apoptosis indices of animals in the ECG group. Combined treatment of ciprofloxacin and KRG may improve the quality of spermatozoa and attenuated apoptosis indices in the ECG group.

Pathogenicity characteristics of an Iranian variant-2 (IS-1494) like infectious bronchitis virus in experimentally infected SPF chickens.

Avian infectious bronchitis (IB) is a major cause of economic loss to the poultry industry. IB virus primarily affects respiratory tract, but strains differ in their tropism for such other target organs as kidneys and alimentary tract. The objective of this study was to estimate the pathogenicity of an Iranian IB virus (IBV) variant (variant-2) which is one of the most prevalent isolates circulating in Iranian poultry farms. SPF chickens were intranasally inoculated with 104 EID50/0.1 ml of the virus. Sera, fecal swabs, and different tissue samples were collected on different days post infection. Clinical signs, gross pathology, and histological changes were recorded. The amount of virus genome was quantified in different tissues and feces using quantitative real-time PCR assay. The highest viral loads were detected in the feces and cecal tonsils. Real-time PCR results demonstrated variant-2 tropism for respiratory tract, digestive system and renal tissue that is due to its epitheliotropic nature. This is the first pathogenicity study of Iranian variant-2 virus. Based on histology observations and clinical signs this isolate was classified as a nephropathogenic IBV. Further knowledge of IBV pathogenesis permits to perform more effective prevention practice.