BACKGROUND: Gallibacterium anatis is a recently defined genus, which is a member of the Pasteurellaceae family. This advantageous pathogen is frequently found as part of the normal microflora of the upper respiratory tract and lower genital tract of the healthy poultry. Provided with appropriate conditions, it leads to various diseases, such as salpingitis, peritonitis, and loss of egg production with mortality in layer flocks. According to previous studies, multiple antibiotic resistance has been observed among G. anatis isolates, which can impose high costs on layer flocks. Due to the lack of the pathognomonic symptoms in the conflicts caused by this bacterium,  not enough comprehensive research has been conducted to date on the condition of this disease in Iran.OBJECTIVES: This study aimed to investigate the infection rates of this bacterium via PCR.METHODS: 295 tracheal swabs were collected from 10-layer flocks. Subsequently, the suspected colonies were isolated and identified with morphological features, differential cultivation, and PCR.RESULTS: 43.72 % of the samples were positive.CONCLUSIONS: The results of this study indicated that laying farms in Iran were infected with Gallibacterium anatis; thus, certain measures should be taken to control the factors reducing the production of layer flocks.

BACKGROUND: Bovine Herpes Virus-1 (BHV-1) belongs to the  Alpha herpesviral family. The virus is the cause of Infectious Bovine Rhinotracheitis (IBR) and Bovine Abortion. In the initial infection, the virus proliferates excessively. Moreover, shedding the virus leads to conditions in the latent phase of the disease. Infectious Bovine Vulvovaginit (IPV ) is the genital form of the disease that represents a genital infection and transmits via pustules and mucopurulent secretions. Exposure to the virus in genital mucosa leads to IPV infection through mating or artificial insemination and the diseases that can be transmitted to healthy livestock by frozen sperm during artificial insemination.OBJECTIVES: Viral contamination of the semen is one of the routes to spread the disease among dairy cattle. Therefore, we investigated the presence of the virus in domestic and frozen imported semen consumed in industrial dairy cattle farms.METHODS: In the present study, 140 frozen straws were collected. After melting each straw, 200 µl of obtained semen was used for DNA extraction, which was done directly on the semen samples and via a Genome Extraction Kit. Subsequently, to ensure the accuracy of the extraction, the PCR technique was done using PRM-1 gene primer. Tracking the viral genome was done using the PCR technique and known primers.RESULTS: In total, one out of 140 samples was found to be virally contaminated, and IBR contamination was confirmed by repeating all the steps and determining the gene sequence.CONCLUSIONS: It is necessary to further investigate the possibility that contamination can be transmitted via frozen semen, given that even one out of 140 samples is contaminated, and the importance of the disease.

BACKGROUND: Clostridium perfringens is an important animal pathogen that causes severe loses to the livestock and poultry industries. Therefore, bacterial detection is believed to be of particular importance. OBJECTIVES: The present study aimed to identify Iranian isolates using conventional and molecular methods and to evaluate their toxicity. METHODS: In this work, 23 Clostridium perfringens type B isolates were examined via microbiological and biochemical tests. Subsequently, they were subjected to PCR technique for the final confirmation. After culturing of the isolates in specific medium, the minimum lethal dose test was performed. The most toxigenic isolate and reference strain was prepared the enterotoxaemia anaculture vaccine. Serum neutralization test was performed on the experimental inactive vaccines. RESULTS: The results revealed that etx and cpb gene could be found in all of the isolates, yet cpb2 gene was found in 65.2 % of the isolates. The minimum lethal dose ranges for these bacteria was less than 1/10 to more than 1/900. The results of serum neutralization in Iranian isolate and reference strains were 5 and 10 IU / ml, respectively. CONCLUSIONS: The findings herein implied that strain 1795 with high toxicity could be used in vaccine production. Of course, for use in production, further research on target animals is needed.

BACKGROUND: Neosporosis is a disease caused by the protozoan Neospora caninum, which is characterized by abortion in cattle and neuromuscular paralysis of various organs, particularly the hind limbs of dogs. The diagnosis of neosporosis is often made by serological molecular tests.OBJECTIVES: This study was conducted to investigate the presence of N. caninum oocysts in the feces of dogs. METHODS: A total of 100 fecal samples were collected from indoor and outdoor dogs during 2018-2019 around Tabriz. Information about age, location, and history of antiparasitic treatment of the dogs were recorded in a questionnaire. Primarily, fecal samples were examined microscopically for Neospora ocysts. After breaking the collected oocysts through freeze-thaw and sonication, DNA contents of the oocysts were extracted and analyzed via PCR.RESULTS: In a light microscopic study, oocysts were observed in 45 (45 %) of the fecal samples. In the PCR study, 21 of the 45 cases tested positive for Neospora infection (21 %). All the positive cases of infection were observed in molecular examination in dogs older than one year. The positive cases were observed in 2 % of the domestic dogs, 8 % of the stray dogs, 6 % of the kennel dogs, and 5 % of the rural dogs. Furthermore, 19 % of the infected dogs had no history of antiparasitic treatment; only 2% had a history of antiparasitic treatment. The results of statistical analysis showed that the rate of infection in dogs around Tabriz with Neospora caninum was significantly (P<0.05) related to the animal's living environment and history of antiparasitic treatment. However, this rate was found to have no significant relationships with the age of the animals.CONCLUSIONS: Due to the high rate of infection with Neospora caninum in dogs in Tabriz, it is necessary to apply preventive methods in traditional and industrial farms around this city and use rapid diagnosis methods in them.

BACKGROUND: Staphylococcus aureus is one of the most important human pathogens that cause infection and also food intoxication by secreting various enterotoxins. Conventional culturing methods to detect S. aureus have some limitations such as being time-consuming due to bacterial growth and low precision and sensitivity in detecting viable but non-cultivable cells. OBJECTIVES: The objective of this study was to detect and quantify enterotoxigenic (A-E) S. aureus in cream pastry products applying PCR coupling with propidium monoazide (PMA) to distinguish dead and live cells. METHODS: One hundred samples were randomly collected from pastry shops in Amol, in a period of 2 months. After preparing dilutions, bacterial pellets were separated and treated with PMA before DNA extraction. Real time PCR was conducted in order to quantify S.aureus cells and enterotoxigenic strains using specific primers. RESULTS: Results of conventional method were close to PMA-qPCR data (P>0.05), but data from qPCR that includes live and dead cells shows more bacterial count than two other methods. Sensitivity of the method applied in the present study, detecting low number of S.aureus cells (less than 10/g) seems considerable. CONCLUSIONS: Findings showed that applying PCR coupling with PMA results in more reliable data than conventional culturing method. Regarding this approach being time-effective, considerably sensitive and specific, it is expected that it be used in food quality control labs in monitoring systems in future.

BACKGROUND: Toxoplasma gondii is an intracellular protozoan parasite that can infect most species of warm-blooded animals, including birds and humans. Because of feeding habits of domestic chickens, prevalence of Toxoplasma infection in free-range chickens is considered as a suitable indicator of environmental distribution of oocysts. OBJECTIVES: The present study was designed to investigate the seroprevalence of Toxoplasma infection in domestic chickens from Khorramabad and compare the results obtained from serological and molecular methods. METHODS: In total, 97 serum samples were randomly obtained from domestic chickens and examined for the presence of anti-Toxoplasmaantibodies using modified agglutination test (MAT). Fifty grams of muscles (mixture of breast and heart) and whole brain from seropositive chickens were separately homogenized and examined by PCR which targets the repeated element (RE) of the parasite. RESULTS: Anti-Toxoplasma antibodies were observed in 21 of 97 (21.64%) sera. T. gondii DNA was detected in 10 out of 21 (47.61%) seropositive chickens (with titres of ≥1:20). The low agreement between serological and molecular results can be explained by several factors such as possibility of cross-reactions in MAT and/or limited sample size in PCR. CONCLUSIONS: These results indicate that domestic chickens may have an important role as a source of infection for cats and individuals living in rural areas.

BACEKGROUND: Toxoplasmosisis one of the most important zoonotic diseases in Iran and the world. OBJCTIVES: Due to the high consumption of lamb meat and the high frequency of Toxoplasma infection in sheep in Iran, the aim of study was to determine frequency of Toxoplasma infection in the slaughtered sheep of Mashhad area. METHODS: In order to do this study, from summer 2015 to spring 2016, 25 blood and 25 heart muscle samples were seasonally collected from Torghabae slaughterhouse in Mashhad area. The samples were transferred to parasitology laboratory. First, the blood samples were centrifuged and the serum samples were isolated, then a portion of the heart muscles sample was taken for PCR examination. The sera and muscles samples were kept at -20 ºC in freezer until examination time. The sera samples were examined to detect antibody against T.gondii by ELISA method. DNA of heart muscle was extracted by commercial extraction kit and was examined to detect Toxoplasma DNA by nested –PCR. RESULTS: In the present study, of 100 sampled sheep, only 1 (1%) of the serum samples was seropositive, while 22 (22%) of the DNA samples were PCR positive. In this study, the highest frequency of Toxoplsma PCR-Positive was seen in spring and the lowest in summer in sheep. Also, the result of this study showed that the agreement between the molecular and EISA method was “fair”. CONCLUSIONS: Based on the high frequency of Toxoplasma infection in heart muscle of sheep, it seems that the risk of transmission of Toxoplasma infection from sheep meat is high.

BACKGROUND: Neospora caninum is a protozoanintracellular parasite which is considered as one of the main factors forrecurrent abortions of dairy cattle in various countries such as Iran. Thisparasite leads to negative economic impacts such as decline in reproduction,reduced amount of milk, and long calving intervals. OBJECTIVES: Therehave been numerous tests to determine the cause of abortion. PCR test isconsidered as a suitable method to specify Neospora caninum DNA and itcan determine the DNA in tissue samples and body fluids of the aborted fetus.This study aims to use PCR to evaluate parasites in the tissues of abortedfetuses so as to detect the best tissue for determining the parasite. METHODS:In this study, 82 aborted fetuses in the first six months of 2015 were studied.The tissues were selected from brain, liver, lungs, spleen, kidneys and rennetfluids. The NP21plus primer was used to detect the presence of Neosporacaninum in samples. After conducting the PCR Test, samples with 340bp bandin Gel electrophoresis were considered as positive. Statistical data from thesurvey of Neospora caninum’s presence in selected tissues were evaluatedby SAS (version 9.2) software. RESULTS: Contamination with this parasitewas found in 34 brain samples (41.5%) of aborted fetuses. In 2 (2.4%) and 4(4.9%) of the aborted fetuses, parasite DNA was found in lung and liver tissuesalong with brain tissues, respectively. CONCLUSIONS: Due to significantdifference of infection of brain tissues in comparison to other tissues, ourstudy considers brain tissue as the most appropriate sample for detecting Neosporacaninum infection in aborted fetuses in PCR method

BACKGROUND: Granuloca cells have a key role during estroeidogenesis. OBJECTIVES: The purpose of this study was to determine the effect of number of culture medium granulosa cells on estradiol concentrations and mRNA codding estrogenic and progestagenic enzyme. METHODS: Briefly, follicles between 2 and 5 mm diameter were dissected from the ovaries of adult cows and were collected by rinsing the follicle walls with Dulbecco Modified Eagle medium/F12 (DMEM/F12). The number of cells was counted with a haemocytometer and the viable cells were assessed by the dye exclusion method using 0.4% Trypan Blue. Treatments were 1) 500,000 cell/500 ml, 2) 250,000 cell/500 ml, 3) 500,000 cell/200 ml 4) 250,000 cell/ 200 ml.  All data were analyzed by JMP (SAS). RESULTS: Low plating density increased E2 secretion and mRNA encoding LHR, FSHR and estrogenic enzymes (17βHSD, CYP19), whereas decreased mRNA encoding GADD45β. There were no differences among treatments for RNA and protein concentration. Low plating density also decreased protein amount but there was no difference among treatments for RNA amount. In conclusion, decreased cell density cause increase in mRNA encoding codding estrogenic enzyme gene expression and decrease in mRNA encoding progestagenic enzyme gene expression. CONCLUSIONS: Protein concentrations did not changed with decreased cell density therefore we can save cells against harmful effect of increasing cell density.

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.