Effect of Vaccination on Distribution and Immune Response of Avian Influenza Virus H9N2 in Coturnix coturnix.

Influenza viruses can multiply in quails and be transmitted to other animal species. As vaccination reduces virus shedding in chickens, the effect of the killed H9N2 avian influenza virus (AIV) on tissue distribution and virus shedding was evaluated in quails. One hundred 20-day-old quails were divided into six equal groups, kept in separate pens, and fed ad libitum. Before vaccination, blood samples were randomly collected from the wing veins. Four groups were vaccinated with the inactivated H9N2 Razi Institute vaccine at 21 days subcutaneously at the back of neck. Three weeks later, two groups were re-vaccinated. Two weeks later, at the age of 56 days, three groups were challenged with 100 µL of allantoic fluid containing 105 EID50 H9N2 through the oculonasal route. Blood samples were collected from quails at 42, 56, 63, and 70 days from each group to determine AIV antibodies by the hemagglutination inhibition test. Three quails were randomly selected and euthanized from each group on days 1, 3, and 6 post-inoculation (PI). Tissue samples were collected, and the RT-PCR test was performed. No clinical signs or gross lesions existed in any of the groups during the experiment. However, the virus was detected in different tissues on the first, third, and sixth days after the challenge in unvaccinated challenged birds. Virus detection was significantly more frequent in the quails vaccinated once and challenged than in the twice-vaccinated challenged group (P≤0.05). On the third day of PI, the virus was detected in some organs of the challenged groups. On the sixth day of PI, the virus was detected only in the lungs of two unvaccinated and once-vaccinated challenged birds. It was concluded that the vaccination of quails against AIV H9 is necessary to protect them from clinical signs, as well as respiratory tract and intestine replication. Two-time vaccination significantly protects the respiratory and intestine tracts, compared to one-time vaccination (P≤0.05).

The role of Newcastle Disease Virus in Broiler Chickens with High Mortality of Kerman Province.

The Newcastle disease virus (NDV) is a member of the paramyxoviridea family and has great significance in the poultry production industry, which spends a huge amount of money every year on prevention and economic loss caused by this disease. A wide range of symptoms, including respiratory and nervous disorders, as well as hemorrhage lesions in the digestive system are observed in this disease. This research investigated the presence of NDV in 10 poultry farms with high mortality and respiratory symptoms in Kerman province, Iran (between January 2020 to October 2020). Tissue samples were collected from mortalities of 10 flocks in different parts of Kerman province and inoculated into embryonated eggs. The NDV was detected in the allantoic fluid by polymerization of partial F gene protein. The virus was positive in the samples of 5 flocks. The results of the phylogenetic analysis also showed that the sequence of isolates was related to genotype II (three isolates) and sub-genotype VIId (two isolates) of NDVs. It was also found that the amino acid sequences of sub-genotype VIId isolates in the 113 to 116 positions were RRQKR and in the 117 positions was the presence of F (phenylalanine). The other three isolates were grouped with B1, Clone, and LaSota vaccines, and the amino acid sequence in the cleavage site included GRQGRL. The similarity between the studied isolates was 99.6%-98.4%. In this study, virulent viruses were isolated and tracked in broiler farms that were vaccinated with live and killed vaccines. It is recommended to pay more attention to designing the vaccination program.

Investigation of Newcastle Disease Virus Infection in Pet Birds in the Southwest of Iran.

Newcastle disease virus (NDV) is a serious threat to the international poultry industry. Therefore, to determine the role of pet birds (Psittaciformes and Passeriformes) in its spread and epidemiology, the presence of this virus in these birds was investigated. In this study, fecal and cloaca swabs from 63 Psittaciformes and 37 Passeriformes, along with tissue samples of dead birds, including proventriculus, trachea, lungs, and intestine, were collected from breeding and sales markets as well as the birds referred to Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran. Isolation of the virus was performed by injecting the suspension of the samples into the allantoic fluid of fertilized eggs, and NDV was detected in the achieved allantoic fluids by reverse transcription polymerase chain reaction. The NDV was detected in 13 allantoic samples. The partial F gene sequences of 10 positive samples were investigated, and their genetic relationship with each other as well as with other isolates in the gene bank was marked. Consequently, subgenotype VII.1.1 (VIId) was in the locus of all 10 viruses. By the amino acid cleavage site sequences of F protein, 10 isolates were determined as velogenic NDV. Moreover, all sequences were similar to each other and other Iranian isolates. Furthermore, the 112RRQKR/F117 pattern was the main amino acid (aa) sequence in the F-protein Cleavage site for VIId genotype isolates.

Avian Influenza-Killed Vaccine on Tissue Distribution and Shedding of Avian Influenza Virus H9N2 in Ducklings.

Ducks play an important role in the transmission of avian influenza to poultry farms. Because of the importance of vaccination in reducing virus shedding, this study evaluated avian influenza-killed vaccine H9N2 on tissue distribution and shedding of avian influenza virus H9N2 in ducklings. One hundred-day-old ducklings were purchased and, after bleeding from 20 birds, were kept in four separate rooms under standard conditions. Groups 1 and 2 were vaccinated at 9 days, and groups 2 and 3 were challenged with 0.1 ml of allantoic fluid containing 105 EID50 (A/chicken/Iran/Aid/2013(H9)) virus intranasally at 30 days. Group 4 chicks were kept as the control group. Chicks were observed two times daily. On days 1, 3, 5, and 8 after inoculation, 3 chicks were randomly selected from each group and cloaca and trachea swabs samples were collected from each bird. Then the ducklings were euthanized and trachea, lung, spleen, intestine, liver, and brain tissue samples were collected for molecular detection. The virus was detected in the tissues and tracheal and cloacal swabs by polymerase chain reaction (PCR), and anti-AIV titres were measured by HI test. The results showed no clinical signs in the challenged groups. In the vaccinated challenged group, virus was detected only in cloacal swabs, but in the unvaccinated challenged group, virus was detected more in tracheal swabs than in cloacal swabs. In challenged-unvaccinated chicks, virus was detected in the trachea and lungs, and in challenged-vaccinated birds, virus was detected in the intestines. In conclusion, vaccinating ducks against the AI H9N2 virus reduced shedding and tissue distribution of AI viruses in challenged ducks.

Molecular identification of aviadenoviruses in broiler chickens suspected to inclusion body hepatitis in Golestan province, Iran.

BACKGROUND: Fowl adenoviruses (FAdVs) are distributed widely throughout the world, and domestic avian species of all ages are susceptible. Fowl aviadenoviruses (FAdVs) can be separated into 5 different species (A-E) with various genotypes and 12 serotypes. Some geno- or serotypes induce hepatitis-hydropericardium syndrome (HPS), inclusion body hepatitis (IBH), and adenoviral gizzard erosion (AGE). AIMS: Detect FAdVs serologically and molecularly and sequencing of FAdVs in broiler flocks in Golestan province. METHODS: From December 2017 to June 2018 liver tissues and blood samples were collected from 31 broiler flocks suspected of IBH. Polymerase chain reaction (PCR) was applied on liver samples and the positive samples were sequenced and antibody against FAdVs was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Out of 31 flocks, the titers of 29 flocks (93.5%) were high in ELISA test for FAdVs and 22 flocks (70.96%) were positive in PCR test. Sequence analysis indicated the isolates belonged to D and E genotype of adenovirus. CONCLUSION: Inclusion body hepatitis caused by FAdVs, are spreading increasingly in broiler flocks of Golestan province and more attention and surveillance programs of breeder and broiler farms are needed to develop preventive measures. Moreover, vaccination of poultry farms in Iran should be considered by more complement studies.

Molecular identification of reovirus in broiler type flocks in Golestan province, Iran.

BACKGROUND: Avian reovirus (ARV) has a global distribution in nature and most clinical signs are found in broiler type chickens. AIMS: This study was conducted to detect and identify reovirus infections from vaccinated breeder chickens and their progenies. METHODS: A total of 20 tissue and blood samples were collected from vaccinated broiler breeders and their progenies with gastrointestinal or performance problems during peak production. Antibody titers were measured by indirect enzyme-linked immunosorbent assay (ELISA) tests. RNA extraction from tissue samples was performed and cDNA was prepared and directly used in the polymerase chain reaction (PCR). Nucleotide sequences were bilaterally determined using internal primers. The analysis of the nucleotide sequences and their related amino acids was performed by the specialized Molecular Evolutionary Genetics Analysis software (6th version). RESULTS: The virus variant was detected in two vaccinated broiler breeders and five broiler flocks. The vaccine strains in breeder flocks included S1133, SS412, 1733, 2408 belonging to genotype 1 from the reovirus phylogenetic tree. Sequence 7 from the isolated reovirus based on the σC revealed that they were different from the reovirus vaccine, and that 6 isolates belonged to genotype 1 of the phylogenetic tree while 1 isolate belonged to branch 4 of the phylogenetic tree. CONCLUSION: The study showed that the new reovirus strain isolated from vaccinated birds differs from common strains used in the vaccines. It is therefore essential to prevent the effects of the field reovirus on the performance of industrial poultry, by updating and inventing new commercial vaccines, live and killed, against the reovirus.