Endocannabinoid and nitric oxide interaction mediates food intake in neonatal chicken.

The aim of the current study was to investigate the interaction of the nitric oxide and cannabinoidergic systems on feeding behaviour in neonatal chicken. A total of 6 experiments were designed to evaluate the interaction between cannabinoidergic and nitrergic systems on food intake in 3-h food-deprived (FD3) neonatal chickens. In Experiment 1, chickens received intracerebroventricular (ICV) injections of saline, 2-arachidonoylglycerol (2-AG) (a CB1 receptor agonist, 2 µg), l-arginine (nitric oxide precursor, 200 nmol) and co-administration of 2-AG + l-arginine. In Experiment 2, ICV injection of saline, 2-AG (2 µg), l-NAME (a nitric oxide synthesis inhibitor, 100 nmol) and their combination (2-AG + l-NAME) were applied to the birds. In Experiment 3, injections were saline, CB65 (a CB2 receptor agonist, 1.25 µg), l-arginine (200 nmol) and CB65 + l-arginine. In Experiment 4, birds received ICV injection of saline, CB65 (1.25 µg), l-NAME (100 nmol) and CB65 + l-NAME. In Experiment 5, chickens were ICV injected with saline, l-arginine (800 nmol), SR141716A (a selective CB1 receptor antagonist, 6.25 µg) and l-arginine + SR141716A. In Experiment 6, birds were injected with saline, l-arginine (800 nmol), AM630 (a selective CB2 receptor antagonist, 5 µg) and l-arginine + AM630. Cumulative food intake was recorded until 2-h post injection. ICV injection of CB1 and CB2 receptor agonists increased food intake. Co-injection of 2-AG + l-NAME increased the hyperphagic effects of CB1 receptors. CB2 receptor-induced food intake was not affected by co-administration of CB65 + l-NAME. l-Arginine decreased food intake and this effect was amplified by co-injection of l-arginine + SR141716A. However; CB2 receptor antagonists had no effect on l-arginine-induced hypophagia. The results suggest that there is an interaction between endogenous nitric oxide and the cannabinoidergic system on feeding behaviour which is mediated via CB1 receptors in the neonatal chicken.

Molecular identification and phylogenetic analysis of Macrorhabdus ornithogaster based on the 18S rRNA gene in companion birds of Tehran, Iran.

Background: Macrorhabdus ornithogaster (MO) is an infectious yeast which can cause acute gastric disturbances in birds. It has a worldwide distribution with a broad host-range of bird species. Aims: Molecular identification and phylogenetic analysis of MO based on the 18S rRNA gene in companion birds of Iran. Methods: A total of 54 stool samples were taken from birds (10 species) suspected of being infected. The presence of MO in collected stool samples was investigated using direct wet mount microscopy. Specific primers were designed to identify the MO 18S rRNA gene by using PCR. PCR products were sequenced and phylogenetic analysis was performed to determine the molecular diversity. Results: The obtained results demonstrated that 44.44% and 59.26% of the samples were diagnosed positive based on the first and second specific primers, respectively. MO was detected in the feces of canary, goldfinch, budgerigar, toucan, and English budge. Phylogenetic analysis revealed that MO sequence data from canaries, finches, and goldfinches had homology with an MO isolated from a German zebra finch. Moreover, MOs from cockatiels, rosy faced love birds, and budgerigars had a high phylogenetic similarity with multiple references, including American budgerigar, Japanese cockatiel, European goldfinch, and German budgerigar. Conclusion: MO exists in many species of Iranian birds, including goldfinches, budgerigars, toucans, and English budgies. As indicated by phylogenetic and polymorphism data analysis, the newly designed specific primers spanning a large portion of 18S rRNA gene of MO, provides additional tool to detect and study the molecular diversity of MO.

Hemagglutinin-neuraminidase Sequence and Phylogenetic Analysis of Two Newcastle Disease Virus Isolated from Chickens in Iran.

Newcastle disease is a highly contagious viral infection affecting many species of birds that can spread fast between poultry houses and cause a heavy economic burden on the poultry industry all around the world. Fusion and hemagglutinin-neuraminidase (HN) protein are important in the pathogenesis of the Newcastle disease virus (NDV). The HN protein is a critical viral protein with multiple functions and plays a key role in the formation of the virulence of NDV. Head of HN protein is responsible for receptor binding, neuraminidase activity. This study aimed to investigate the sequence homology of hemagglutinin-neuraminidase of two NDV isolates sampled from infected farms in Iran. The samples were collected from flocks that had been vaccinated by both types of live and killed vaccines for NDV. After isolation of NDV, the viruses were subjected to the polymerase chain reaction (PCR) amplification using two pairs of specific primers designed for the HN gene to amplify the complete HN gene (1730bp). Afterward, the PCR products were sequenced and analyzed by phylogenetic tree construction software. Based on the analysis, substantial sequence homology among Iranian isolates is within the range of 97.1-100%. Moreover, the sequence homology searching revealed a level of similarity between HN sequences of Iranian isolates and the HN sequences from other countries, particularly Asian ones. For instance, a high homology ratio (95.34%) was found between Iranian isolates and the sequences registered on online molecular databases from China. Based on phylogenetic analysis, the NDV isolates belong to the VIId genotype. Finally, it can be concluded that monitoring the circulation of NDVs among poultry and other birds can help to obtain an insight into the evolution of NDVs and control of panzootic viruses in the future.

laying Farm: Up to Date Data on a Fowlpox Outbreak in Phylogenetic Analysis in Iran, 2018.

Fowlpox is an economically significant viral disease in poultry, characterized by two forms of clinical signs, including cutaneous and diphtheritic lesions. This infection can have several adverse effects on flock performance, such as a reduction in egg production and growth and an increase in mortality. In winter 2018, an infection suspected to fowlpox was reported from a Hy-line W-36 laying farm in Isfahan province, Iran. The birds were 38 weeks of age and showed obvious diphtheritic signs in mucous membranes with increased mortality and reduced egg production. In total, 20 samples were collected from diphtheritic lesions (Trachea and Esophagus) of infected birds. The Polymerase Chain Reaction method was used to amplify a 578 bp fragment of the poxvirus 4b core protein gene. Phylogenetic relationships of avian poxviruses are usually analyzed using the 4b core protein-coding gene sequences with molecular weights of 75.2 kDa. The major elements had the fowlpox genome, and sequencing was performed for one isolate as representative. The nucleotide sequence result showed that this isolate (FP\UT-POX-2018) had a similarity rate of 99.53% with the previous Iranian fowlpox isolate (FP\GHPCRLAB.3) sequenced in the GenBank.Moreover, there was a 100% similarity among the current isolate nucleotide sequence, FP/NobilisVarioleW, and FP/FPV-VR250. The derived phylogenetic tree showed that these isolates were clustered in A1 subclades. Therefore, Iranian isolates of fowlpox virus have remained in the same subclade of phylogenetic classification (subclade A1), and they show high genomic similarity with previous isolates of Iran. Veterinarians and farmers must not underestimate fowlpox. However, they should consider the importance of vaccination against this disease like any other disease care.

Molecular Detection of Mycoplasma synoviae from Backyard and commercial Turkeys in Some Parts of Iran.

M. synoviae (MS) is an economically important pathogen and the major cause of airsacculitis and infectious synovitis in turkeys. Infection with this pathogen may remain asymptomatic but can render infected birds susceptible to secondary infections. This study was carried out for the molecular detection of MS infection in commercial and backyard turkey flocks in Tehran, Semnan, Isfahan, Qazvin, Zanjan, East Azerbaijan, Gilan, Mazandaran, and Golestan provinces of Iran. Sixty-hundred tracheal, choanal cleft or/and infraorbital sinus samples were collected from 18 commercial and 31 backyard turkey flocks. The polymerase chain reaction (PCR) technique was performed by using primers specific for detecting the 16S rRNA and vlhA genes of MS. The results showed that 51.61% of backyard and 33.33% of commercial farms were MS-positive. These findings suggested the molecular presence of MS, especially in northern and central regions of Iran. Further, the frequency of MS-positive samples was significantly lower in commercial farms than backyard farms (P<0.05).

A survey of the prevalence of infectious bronchitis virus type 4/91 in Iran.

To evaluate the prevalence of infectious bronchitis virus (IBV) type 4/91 in Iran, tracheal swabs from 77 broiler flocks in 16 provinces were collected at the slaughterhouse. Swabs were subjected to RNA extraction and tested by RT-PCR, followed by a type-specific nested PCR. The viral RNA was detected in 33 samples (42.8%) from different provinces. The results indicate a relatively high prevalence of IBV type 4/91 in Iran and necessitate revising the vaccination programme against this disease.

Isolation and identification of Ornithobacterium rhinotracheale in broiler breeder flocks of Guilan province, north of Iran.

The aims of the present study were to isolate and serotype, determine the Seroprevalence, Drug susceptibility and diagnosis of infection by Polymerase Chain Reaction (PCR). In this study 460 serum samples and 220 tracheal swabs, 90 ovaries and oviduct swabs, 90 misshapen egg shells swabs were collected from 22 broiler breeder flocks of 5 companies. Serological results showed that all of the 22 flocks (100%) were positive for ORT infection. Ornithobacterium rhinotracheale (ORT) antibodies were detected in 289 (62/83%) out of the 460 serum samples. ORT was detected from tracheal swabs of seven flocks (31/81% or 3/18% out of 220 tracheal swabs). There was significant correlation between flock different ages and ORT titers (p<0.05), but correlation of flock ages and ORT isolates was not significantly different (p>0.05). Seven flocks infected with ORT were detected positive in PCR but bacteria were Isolated from only five culture. No ovaries and oviducts, misshapen egg shell swabs yielded ORT. A 784 bp fragment of the 16S rRNA gene was amplified using ORT specific primers in the PCR. All the isolates were identified as serotype A by Rapid Agglutination Test. Drug sensitivity test using standard disk diffusion technique was performed with 27 antibiotics. Antibiotic susceptibility for Quinolons family was seen more than the others and Cephalosporins family except to Cephalexin. The isolates were 80-100% susceptible to Tetracycline family and the most antibiotic resistant were seen for Aminopenicillins, Polypeptides, Sulfanamides and 80-100% resistant to Aminoglycoside family. Eighty percent of the isolates were resistant to Licomycin and 60% were moderate sensitive to Lincomycin. This study is the first report of prevalence of ORT, bacterial isolation, biochemical characteristics, serotyping and molecular method (PCR) in broiler breeder flocks in Guilan province of Iran.