m. mayahi

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).

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 tonsils, 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.

Newcastle disease, as one of the most important diseases of the poultry industry, has many economic losses. The aim of this study was to investigate the role of Newcastle disease virus in respiratory syndromes of broiler flocks in Khuzestan and Isfahan provinces. For this purpose, tissue samples (trachea, lung, and cecal tonsils) were taken from ten dead birds of 20 broiler farms with respiratory symptoms in different regions of these two provinces. Samples were inoculated into 9 to 11 days old embryonated chicken eggs. The RT-PCR was performed to detect Newcastle disease virus on allantoic fluid. Out of 20 studied flocks, the virus was isolated and identified in nine flocks. The 330 base pair nucleotide sequence in the F gene virus cleavage site was examined in 9 isolates. The results of RT-PCR test showed that Newcastle disease virus was isolated from respiratory syndromes in 50% of broiler flocks in Khuzestan province and 40% of broiler flocks in Isfahan province. Of the 9 isolates, 3 were in genotype II and 6 were in genotype VIId of class II Newcastle disease virus, and isolates of both genotypes were present in both provinces. The variety of VIId isolates found in Khuzestan province was more than Isfahan province. Isolates belonging to genotype II in this study were similar to Pakistani, Chinese and Nigerian strains, and other isolates were similar to previous Iranian strains.

Avian pasteurellosis (fowl cholera) is an important disease affecting domestic and wild birds all over the world. Although the capsular type A of Pasteurella multocida is mostly involved, other capsular types are occasionally incriminated. The present study aimed at investigating the effect of some adjuvants on immunogenicity and protectivity of P. multocida bacterin in chickens, compared to an Iranian commercial vaccine. Eight-week-old chicken pullets were double vaccinated with an interval of three weeks. Vaccine immunogenicity testing was conducted using an in-house indirect enzyme-linked immunosorbent assay and assessing serum antibody titers at 7, 14, and 21 days post-primary and 14 days post-secondary immunization. The possible adverse effects were recorded by a poultry-disease expert. For evaluating the vaccine protection rate, chickens were subjected to 2×Lethal Dose 50%of a virulent P. multocida strain two weeks post-secondary immunization. The rate of live and normal animals was regarded as protection rate 7days after the exposure. The findings showed that oil adjuvants Montanide ISA 70-and Montanide ISA 71-containingvaccines (with or without saponin) caused a powerful immune reaction than the aluminum adjuvanted vaccine and commercial vaccine (P<0.05). Significant protection against challenge was merely induced by the oil adjuvanted vaccines (P<0.05). The majority of the studied chickens showed inflammation at the injection site (yellow) throughout the trial. Vaccines made by Montanide ISA 70 and Montanide ISA 71 are novel and effective inactivated vaccines that are able to cause significant protection to fowl cholera disease.

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.

Turkey Astroviruses (TAstV) is one of the key factors in the emergence of the enteritis syndromes in poults. The aim of this study was to investigate the astrovirus contamination of some industrial turkey flocks with diarrhea in Zanjan province of Iran by RT-PCR and then to determine the genome sequence of the detected viruses. For this purpose, Samples were collected from 30 suspected turkeys' flocks less than 3 weeks ages with clinical signs, diarrhea or fecal pest around vent during 2018-2019. From each suspected flock, randomly ten turkeys were collected and after euthanized, post mortem performed and probable gross lesions were noted in special work sheet. The RT- PCR was performed. The analysis of the nucleotide sequences and their related amino acids was performed. The virus was detected in eight (66/26%) out of 30 flocks.The nucleotid sequences obtained in this study were compared with the sequences in the gene bank and the phylogenetic tree was drawn, ased on the polymerase genes and amino acid sequences obtained from GenBank all eight isolates obtained in this study were in the branch of Astrovirus turkey 2 ( TAstv). These strains most closely resembled the Polish Astrovirus turkey 2 strains.It was concluded that astrovirus play roles in turkeys flock enteritis and farmer should consider prevention and control of turkey's intestinal diseases.

Adenoviruses are common infectious agents in poultry around the world and show a wide range of clinical signs and symptoms. The importance of the presence of adenovirus infections, in addition to economic losses, is associated with other viral diseases.The purpose of this study was the molecular detection of Fowl adenoviruses and histopathological  examination of liver lesions in broiler chickens suspected of inclusion body hepatitis in Markazi and Ghom provinces, Iran. In this regard, the liver samples with macroscopic lesions were collected from dead birds of ten broiler chicken flocks of these provinces in the winter of 97 and spring 98. The chickens of these flocks were at the age of 6 to 41 days. For molecular analysis, PCR was performed on L1 (Loop1) region of the hexon gene. For histopathology, a portion of the liver tissues were placed in 10% formalin. After routine procedures in the pathology laboratory, microscopically examined. Eventually, 9 flocks from the 20 flocks were positive in the PCR test. These positive products were sequenced. These adenoviruses belonged to the D and E genotypes and D11 and 8b serotypes of avian adenoviruses. Also, in the microscopic study of the affected liver, necrosis with several heterophilia infiltration around these regions and within the liver sinusoid was observed. In the nucleus of healthy hepatocytes around the necrosis regions, the inclusion of intra-basophilic nucleus was observed. Confirmation of the presence of the IBH agent in these provinces, the need for prevention programs in mother and broiler farms should be considered.

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).

Detect FAdVs serologically and molecularly and sequencing of FAdVs in broiler flocks in Golestan province.

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).

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.

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.

Present study was designed in order to molecular confirmation, capsular typing and detection of 12 important virulence factors include ompH, oma87, sodA, sodC, hgbA, hgbB, exBD-tonB, nanB, nanH, ptfA, pfhA and toxA in 18 Pasteurella multocida avian isolates from different areas of Iran with a strain of the organism was isolated from the epidemic pasteurellosis in the Northern region, and also survey of acute isolates lethality in poultry embryos and pullet. Molecular confirmation of tested isolates was performed using specific primers of kmt1. As well, capsular typing and detection of virulence factors of isolates were performed by application of multiplex PCR method and using specific primers of capsule biosynthesis genes and genes encoding virulence factors of organism. All isolates were confirmed molecularly but based on the results of capsular typing, 16 (88.8%) isolates with strain isolated from epidemic pasteurellosis were identified as type A and 2 (11.1%) as untypeable isolates. Detection of virulence genes showed that all studied isolates has six virulence genes of sodC, hgbA, hgbB, nanB, nanH and ptfA. The oma87, exBD-tonB, ompH, sodA and pfhA genes were respectively present in 94.4, 94.4, 83.3, 66.6 and 27.7% of the isolates, but there was not any toxA positive isolates. To determine the chicken embryo lethal dose 50 (CELD50) and lethal dose 50 (LD50) of the isolates, different dilutions of five acute isolates containing the majority of virulence genes as well as strain isolated from epidemic pasteurellosis were respectively inoculated to embryonated chicken eggs and lying pullets. Based on the results of CELD50, though all six isolates were able to waste the poultry embryos, but strain isolated from epidemic pasteurellosis was recognized as the most acute isolate. Results of LD50 test showed that none of the five acute isolates were able to waste the studied pullets while the LD50 of strain isolated from epidemic pasteurellosis was determined 5 × 101 colony forming unit per bird (cfu/case), and this strain was identified as the most acute understudy isolate. The findings of this study showed that, despite the presence of the majority of virulence genes in P. multocida avian isolates in Iran, and that the most acute isolates were able to waste the poultry embryos but they were not infectious to lying pullets, and virulence of strains isolated from birds with hyperacute form of the disease is far higher. Our data could be useful in the design of new and efficient vaccines that would be acceptable protection in animal populations.

Î II prell.:nt against reecnt virulent strains or in/ectious bursal disease virus (IBDY), a local isolaLe of the virus was propagated in bursal tissue, l(lfIllali/.ed and used I(lr vaccine production. An experimental vaccine adjll\anted hy oil ISA-70 was prepared and compared with a commercial IBDY-NDY inactivatcd vaccine. A single injection or the two vaccines proteeted chickens against mortality hut the oil adjuvanted bursal derived vaccine conli:rred a higher percent or bursal protection.