BACKGROUND: Newcastle disease is one of the most serious viral diseases in the poultry worldwide. OBJECTIVES: Since the traditional strategies have been hardly effective in controlling the disease, the purpose of this study was to introduce new methods for early and rapid diagnosis of Newcastle. The present study helps to reduce further damage to the poultry industry. METHODS: RNA extraction was performed, using RNease mini kit, according to the manufacturer’s instructions. Extracted RNA with 68.23×109 copy numbers was prepared as serial dilutions of 100 μL for RT-PCR and RRT-PCR reactions. RRT-PCR and RT-PCR were performed, using commercial kit and RNease mini kit, respectively.  RESULTS: Results showed that amplification was done according to prepared dilution equal 10-34 for RRT-PCR reaction and a visible band observed on 1.5% Agarose gel up to 10-20 for RT-PCR reaction. Based on the results observed, RRT-PCR and RT-PCR reactions are able to detect 10-34 and 10-20 copy numbers of primary sample, respectively. CONCLUSIONS: The sensitivity of RRT-PCR reaction is almost twice compared with RT-PCR reaction, also RRT-PCR reaction is able to diagnose Newcastle disease virus in infected samples with 10,000 copy numbers of the RNA virus less than RT-PCR.

BACKGROUND: Causing site direct mutation can be one of the efficient methods to evaluate the characteristics and properties of various genes. Brucellosis is the most common zoonotic infectious disease that would cause great economic losses. Thus, recognition of pathogenic and immunogenic factors in the genus Brucella can lead to control this health problem. Objectives: Considering the importance of site direct mutation in identification of genome structure and numerous ways to achieve this goal, Overlap Extension PCR is introduced as an improved technique for the removal and replacement of the gene target. Methods: For this study, with two-step PCR using specific primers, upstream and downstream fragments from target gene and antibiotic resistance cassette from plasmid pET28a (+), were reproduced and were connected to each other. The resulting fragment was cloned in specific position of pBluescriptIISK(-) plasmid by the restriction enzymes. Then, the construction was transferred into the genome of Brucella abortus by electroporation method. Results: Fusion PCR product was obtained without any change in the nucleotide sequence and then it was cloned into pBluescriptIISK (-) plasmid, finally the construction was replaced and the target gene was deleted. Conclusions: The results of this study show that the Overlap Extension PCR is an optimized and modified technique to create mutations in the bacterial genome structure and can easily be used in the family Brucella.

BACKGROUND: One of the most important heart diseases in dogs is valvular insufficiency, which can be evaluated by diagnosis ways such as phonocardiography, echocardiography, etc. OBJECTIVE: The purpose of the current study was to evaluate of valvular insufficiencies with phonocardiography and echocardiography and using phonocardiography technique in detection of cardiac valvular disease in practice. METHODS: This survey was done on 180 five-year-old dogs which 30 of them had valvular insufficiency. They have been referred to radiology section and echocardiography technique was used after listening to heart sounds and recording heart murmur and surveying by phonocardiography. The type and location of valvular insufficiency was diagnosed by phonocardiography and then echocardiography was used, the results from both techniques was compared afterwards. RESULTS: In all of these 30 dogs, murmur was systolic and mitral insufficiency and mitral regurgitation were diagnosed by phonocardiography. using echocardiography, the mitral insufficiency was confirmed in 28 dogs, one of them has been diagnosed to have tricuspid inssufiency and pulmonary stenosis in addition to mitral insufficiency. In two cases no abnormality sign has been detected. CONCLUSIONS: According to this study, it is recommended to use phonocardiography technique in order to pre-diagnose the valvular insufficiency, it's type and location and use echocardiography to determine the process of disease and control this progress.

BACKGROUND: Brucellosis is one of the most important zoonosis that is prevalent among human and animal. Today, a large percentage of animal and human population suffer from its side effects. OBJECTIVES: The purpose of the present study was to estimate the prevalence of anti-Brucella antibodies in flock- and animal-level in districts of south of Kerman province. METHODS: In this cross-sectional study, 300 herds of 7 districts in the area were selected randomly; 10 samples of sheep and goats in each flock were randomly selected. Out of 3000 samples, 2952 samples were examined using Rose-Bengal test; Wright and 2-ME tests were done on positive samples. Descriptive statistics and logistic regression in Stata 11.2 were used to analyze the data. RESULTS: The seroprevalence of anti-Brucella antibodies in animal- and flock-level was 6.4 and 25.3 percent, respectively. The chance of being infected in sheep was 2.12 times of goats. CONCLUSIONS: The prevalence of Brucella was considerably high in animal- and herd-level in this area. It is necessary to empower Iran Veterinary Organization in financial aspects to control this infection.

BACKGROUND: Adiponectin is one of the most important adipocytokines that regulate male and female fertility via AdipoR1 and AdipoR2 receptors. Recently expression of adiponectin system and its negative regulatory role on hypothalamus-pituitary axis have been confirmed. Objectives: No information is available about the expression pattern of adiponectin and its receptors in hypothalamus-pituitary axis in domestic animals. Here for the first time, we studied hypothalamus-pituitary adiponectin system gene expression in different stages of bovine estrous cycle. Methods: Anterior pituitary and hypothalamus were collected from Holstein cow at the local abattoir. The estrous cycle was classified to four phases (proestrous, metstrous, early luteal and late luteal) based on macroscopic examination of ovaries and uteri. Gene expression analysis of adiponectin and its receptors was done using quantitative real time PCR (qPCR Probe MasteKit) and according to the comparative 2-ΔΔCt method. E2 and P4 levels were measured using ELISA method. Results: Our results demonstrated that adiponectin and its two receptors were expressed in pituitary and hypothalamus of cyclic cow. Maximal expression of adiponectin was observed in early luteal phase, while it was expressed at minimal level during the proestrous stage. We observed no significant changes in the expression of AdipoR1 in both tissues at different stages of estrous cycle. The highest expression of AdipoRII in both tissues was detected during the proestrous stage, while it expressed at minimal level during the late luteal phase. E2 and P4 had respectively negative and positive correlations with adiponectin expression levels in hypothalamus and pituitary. Conclusions: Based on our results that demonstrated adiponectin was minimally expressed at proestrous stage and other data about the negative action of adiponectin on LH secretion from pituitary, we concluded that adiponectin may has role in the hormonal function of this axis during the estrous cycle.

BACKGROUND: Reptiles, especially turtles that inhabit both on land and water, have made some special adaptations. Many people keep turtles as pets. Therefore, the anatomical knowledge of turtles should be more carefully evaluated and used for therapeutic purposes. One of these turtles is European pond turtle (Emys orbicularis). Most of vital systems are enclosed by the carapace and the plastron so it cannot be examined customarily by clinicians. The noninvasive diagnostic imaging techniques provide detailed information concerning these organs. OBJECTIVES: This study was conducted to give complete topographic information and knowledge about the position of the non respiratory organs of the coelomic cavity in the European pond turtle using Computed Tomography (CT) and usual anatomic methods. METHODS: 10 adult turtles (5 female, 5 male) were selected. All scans were obtained on a two detector scanner. In anatomical study three female and three male turtles were dissected. Two other female and male turtles were sectioned transversely. RESULTS: The results showed some differences in the position of the organs including stomach, gall bladder, liver and heart with those of other species. Moreover, the topography of the organs is described in retracted and protruded neck in this article. Retraction of the neck had an influence on the position of the organs such as oesophagus, stomach, liver and heart. CONCLUSIONS: The general morphological features of the non respiratory organs of the coelonic cavity of European pond turtle were examined by CT images and macroscopically in this study. Significant differences were found compared with other species.

BACKGROUND: Pre-purchase radiographic examinations of horses are very important for the diagnosis of possible disorders.OBJECTIVES: This study aims to determine the variation and frequency of common disorders of the tarsus in pre-purchase radiographic examinations of horses and assess the effect of age, sex, and type of hind limb (right or left) on the occurrence of these disorders.METHODS: In this study, we used the radiographs taken from the tarsal joints of the right and left hind limbs in 110 horses for a two-year period which were requested for pre-purchase radiographic examinations. Among these radiographs, the cases that included the standard position of the tarsal joint were evaluated for the existing disorders and the effect of age, sex, and type of involved hind limb.RESULTS: The most frequent disorders were osteoarthrosis (32.27 %), osteochondritis dissecans (5.91 %), and soft tissue swelling (1.36 %). The least common disorders were calcification, desmopathy, and tenosynovitis, each with a prevalence of 0.45%. Also, most of the tarsal ​​joint disorders had a mild degree of severity (80 %). The prevalence of disorders was 100 % for horses aged <4 years (4 out of 4) and 49.1 % for horses aged >4 years (52 out of 106). No significant difference was observed in terms of gender (P=0.65) and type of involved hind limb (P=0.17).CONCLUSIONS: Considering the prevalence of tarsal joint disorders in horses, pre-purchase radiographic examination can be very important for predicting the future health and performance of the horse. Therefore, radiographic evaluation of the tarsal joint should always be a part of pre-purchase examinations in horses.

BACKGROUND: The bacterium Capnocytophaga canimorsus is a relatively newly recognized gram-negative, facultative, slow-growing bacillus that forms part of the normal oral flora of dogs and cats. Considering the pathogenicity of this bacterium in humans, determining its prevalence is very important for public health as well as the health of dog owners.OBJECTIVES: This study aims to investigate the prevalence of Capnocytophaga canimorsus in the normal oral flora of healthy dogs.METHODS: After taking samples from the saliva of 32 healthy dogs without oral, dental or digestive diseases at different ages, breeds, and sexes, they were placed in a test tube containing 10 mL of sterile peptone water with sterile plastic brushes, and immediately sent to the bacteriology laboratory under sterile conditions. The samples were cultured on a chocolate agar medium containing 5 % defibrinated sheep blood. Then, all the samples were kept in a greenhouse for 48 hours at a temperature of 37 °C and under anaerobic conditions. Using a loop, the grown pink colonies were isolated and to confirm the identification of the isolates, polymerase chain reaction (PCR) test was used in three main steps: Gene extraction, PCR reaction, and electrophoresis.RESULTS: Out of 32 saliva samples, four positive cases of Capnocytophaga canimorsus bacteria were identified by PCR diagnostic method.CONCLUSIONS: Given that Capnocytophaga canimorsus bacterium is present in the oral flora of healthy ​​dogs, dog owners should have sufficient and favorable knowledge about this bacterium and related diseases. The PCR method can be used to detect this bacterium.

BACKGROUND: Coronaviruses, which mainly cause gastrointestinal and respiratory infections, have been identified in various species. Among the extensive genomic data of disease-causing Coronaviruses in humans and animals, some similarities can be analyzed by in-silico methods.   OBJECTIVES: In the present study, comparative genome analysis of medical and veterinary medicine Coronaviruses was performed to obtain more accurate information about the genetic similarities and differences of different members of this family.METHODS: The genomic sequences were retrieved from NCBI and Virus Pathogen Resource databases. Using the NCBI database blast algorithm, all sequences were aligned with the SARS-CoV-2 genome sequence, and similarity was obtained. Amino acid sequences of structural and non-structural proteins associated with coding regions (CDS) were aligned separately with the SARS-CoV-2, and their similarities were calculated. The 3D structure from each protein was compared with the corresponding protein in SARS-CoV-2, and Template Modeling Scores (TM-Score) were obtained. A phylogenetic tree of different species of the Coronaviridae family was drawn based on nucleotide and amino acid sequence data.RESULTS: Nonstructural coding gene sequences detected the highest interspecies similarities in nucleotide, amino acid sequence, and 3D structure (nsp12, nsp13, nsp14, and nsp16). The ORF1ab, encoding non-structural proteins, carries essential functions for viral replication.CONCLUSIONS: This study showed that the transcription complex is highly conserved among human and animal Coronaviruses. A comparison and analysis of the Coronaviridae transcription complex can be considered a key target for diagnosing, developing antiviral therapies, and designing vaccines.

BACKGROUND: Respiratory pathogenic beta-hemolytic streptococci in horses, including Streptococcus equi subsp. equi, the causative agent of strangles disease, Streptococcus equi subsp. zooepidemicus is an important cause of respiratory disease and Streptococcus dysgalactiae subsp. equisimilis has been isolated from nasal swabs taken from horses with a history of respiratory disease.OBJECTIVES: The present study aimed to determine the frequency and risk factors of respiratory tract infections originating from beta-hemolytic streptococci in the provinces of West Azerbaijan, East Azerbaijan, and Ardabil.METHODS: During this study, 121 horses with clinical respiratory symptoms were sampled. After performing clinical examinations and recording clinical signs in special worksheets, sampling of the upper part of the respiratory tract was performed using nasopharyngeal swabs. The samples were sent to the laboratory in a standard transfer medium with cold chain.RESULTS: In this study, out of 121 samples collected from horse breeding clubs from 10 different regions of northwestern Iran, 51 were negative for beta-hemolytic streptococci while the results were positive for the other 70 samples (P<0.001). Regarding the positive samples for beta-hemolytic streptococci, the results of differential cultures were as follows: eight cases of Streptococcus equi subsp. equi, 57 cases of Streptococcus equi subsp. zooepidemicus, and five cases of Streptococcus dysgalactiae subsp. equisimilis. There was no significant relationship between the frequency of beta-hemolytic infections with variables of gender, race, and geographical area (P>0.05). Meanwhile, the statistical test showed a significant relationship between the frequency of infection with these bacteria and the variable of clinical symptoms (P<0.001). Moreover, the frequency of beta-hemolytic streptococcal infections was significantly associated with age (P<0.05).CONCLUSIONS: The results herein suggested that the bacterial cause of the majority of respiratory infections in infected and sampled horses in the provinces of West Azerbaijan, East Azerbaijan, and Ardabil at the time of sampling was Streptococcus equi subsp. zooepidemicus and that this organism is a potential pathogen for respiratory diseases in horses in these provinces.