BACKGROUND: Sheep are a form of investment and a quickly liquidatable resource, particularly in traditional and low income production systems. Tropical and long warm-season regions always affect sheep production negatively. Methods: In this experiment 15 female and 7 male sheep were chosen and their body temperature, heart rate, respiration rate measurements and blood sample for biochemical parameters analysis were taken during May 5 to September 5. Results: Heart rate and respiration rate in male sheep were a little higher compared with female sheep but there was no significant difference between them respiration (56 vs. 55) and beat (120 vs. 118 per min). Rectal temperature wasn’t significant between two sexes (40.6-40.09 C˚). Also skin temperature wasn’t significantly different between two sexes (36.02- 36.08 C˚). The only difference was related to month effect (p<0.05). Sex and month hadn’t significant different effects on blood urea, creatinine, glucose and potassium concentration. Blood urea concentration of female sheep was not significantly higher than male’s (p>0.05). Blood Sodium concentration was significantly different between two sexes so that male sheep had the highest minimum average (p<0.05). There was no significant increase in blood Potassium concentration of female sheep compared to male sheep (p>0.05). ConclusionS: These results indicated that sheep of this region had been well adapted to summer heat stress and they showed usual changes of blood metabolites in response to heat stress.

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: Fibroblasts are one of the important cells in wound healing. These cells create a proper bed for keratinocytes migration and wound contraction.Wound healing in distal limb of horses has complications, such as formation of exuberant granulation tissue (EGT). The main factor in this problem is overgrowing of fibroblasts. ObjectiveS: The purpose of the present study was to compare fibroblast growth curve in isolated skin from horses’ neck and distal limb. Methods: 5 horses with normal hematological and clinical signs were selected. Two samples of full thickness of skin were taken from the neck and lateral metacarpal region of each horse asseptically. Then the samples were washed with PBS minced and placed in ventilated flask 25 cm2. After attaching samples to flask, 5 ml culture medium(RPMI-1640 with 10% FBS) were added and the flask was placed in an incubator at 37°c in 5% CO2. After leaving a sufficient number of cells from tissues adhered to the bottom of the flask, the cells were passaged to a new ventilated flask. After growth and proliferation of cells, they were passaged again and a suspension of cells in culture medium (10000 cells/ml) was maked. To each cell of a 24-well plate, one ml of this suspension was added. After 48 hour, cells of 3 well were detached with tripsin daily, counted and viability determinted within 8 days. Results: There was no significant difference between viable cells number but there was significant difference in viability percent of cells in neck and distal limb. The mean of population doubling time (PDT) for fibroblasts of neck is 31.73 hours and for fibroblasts of distal limb is 26.4 hours. This difference was not significant. ConclusionS: With regard to different viability percentage, it seems that the appoptosis in fibroblasts of neck skin is more regular than distal limb skin.

BACKGROUND: Probiotics, in form of microbial supplements, are known to be a suitable alternative for antibiotics and can affect the health indicators of host. Objectives: The present study was conducted to assess the combined effects of dietary autochthonous Saccharomyces cerevisiae and Aspergillus niger on haematological and serumbiochemicalparameters of beluga sturgeon (Huso huso) juveniles. Methods: This study was based on a completely randomized design with 4 treatments and 3 replicates on beluga juveniles with average weight of (mean ±SE) 31.8±2.81g. Beluga Juveniles were divided randomly into 12 fiber glassy tanks with density of 30 fish per tank and were fed with diet contain dietary probiotic with density of 2×106 (Cells/g) for the first treatment, 4×106 (Cells/g) for the second treatment, 6×106 (Cells/g) for the third treatment and basal diet without probiotic for the control group for 8 weeks. Results: Diet supplementing with concentration of 6×106 (Cells/g), significantly improved serum biochemical parameters (p<0.05), however hematological parameters were affected by supplemented diet with probiotics that showed no significant difference in comparison with the control group (p>0.05). Also results indicate that growth factors were improved in experimental treatments in comparison with the control group. Conclusions: The results showed that the use of combination of these species with studied concentrations can improve the performance of some biochemical parameters such metabolites factors, immune, enzymes and serum electrolytes of belugajuveniles. It is recommended that the concentration of A. niger and S. cerevisiae, used for third treatment be used as an immune stimulator for beluga juveniles.

BACKGROUND Honey bee venom contains complex compounds such as polypeptides, enzymes, and amines. One of the important components of bee venom is the phospholipase A2 enzyme, which is considered an important honey bee venom allergen and is also used to treat some diseases. This enzyme is found in other insects, arachnids, snakes, and mammalian cells, and its function is the hydrolysis of the second ester bond of glycerophospholipids and the release of fatty acids and lysophospholipids. Although transient transfection can produce recombinant proteins, stable cells are more suitable for high-scale production with economic efficiency.OBJECTIVES: The present study created a stable cell line to produce recombinant phospholipase A2 from honey bee (Apis mellifera) venom.METHODS: Plasmid cloning DNA vector containing phospholipase A2 gene was prepared by Macrogen Company. The recombinant plasmid was transferred to Chinese hamster ovary cells by heat shock method, and gene expression was carried out in a HamsF12 culture medium containing neomycin antibiotic. After increasing polyclonal strains containing plasmid, monoclonal clones were selected by limiting dilution. Then, monoclonal clones were propagated, the soup of the selected cells was collected and concentrated, and the protein expression was checked by sodium dodecyl-sulfate polyacrylamide gel electrophoresis test.RESULTS: The results of electrophoresis, which was performed to confirm the expression of the phospholipase A2 gene in the cell soup, showed a band with a molecular weight of 20 kilodaltons, which confirms the creation of a stable cell line for the production of recombinant phospholipase A2 honey bee venom.CONCLUSIONS: After the transient transfection of the plasmid containing this gene, several cells undergo recombination due to having repair mechanisms and putting the desired gene along with the antibiotic resistance gene in their genome. These cells can be selected and propagated by adding antibiotics to the culture medium.

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: Infectious diseases and microbial antibiotic resistance are the major problems of fish farming industry annually causing remarkable losses. Apart from the economic losses caused by these infections, some of these agents are zoonotic and may be transmitted to humans.OBJECTIVES: This study was aimed to identify the common causative agents of infections in rainbow trout farms and to determine their antibiotic resistance toward some common antibiotics.METHODS: Sampling was performed during a nine-month period between March and December 2021 by visiting and inspecting rainbow trout farms and the affected fish with disease symptoms were obtained from the farmed fish in Mazandaran, Lorestan, Chaharmahal and Bakhtiari and Zanjan provinces. Bacterial culture was undertaken from anterior kidney or spleen organs and the isolated bacterial strains were identified by phenotyping, biochemical and molecular assays. Antibiotic resistance pattern was evaluated by disk diffusion method (DDM) and minimum inhibition concentration against erythromycin, oxytetracycline, florfenicol, enrofloxacin and nitrofurantoin.RESULTS: Seventy-four bacterial isolates of Gram-positive cocci or Gram-negative coccobacilli were isolated. In phenotyping, biochemical and molecular (PCR) assays Lactococcus garvieae (12 isolates, 16.2 %), Aeromonas hydrophila (9 isolates, 12.2 %), Streptococcus iniae (17 isolates, 23 %), Streptococcus agalactiae (20 isolates, 27 %), and Yersinia ruckeri (16 isolates, 21.7 %) were identified. The majority of these isolates were obtained from the fish farms in Mazandaran province. Erythromycin and oxytetracycline with 87.8 % resistance were antibiotics with the highest resistance, while enrofloxacin with 24.3 % resistance revealed the lowest level of resistance. Antibiotic resistance rates for florfenicol and nitrofurantoin were also 43.2 % and 44.4 %, respectively. The highest antibiotic resistance was detected in the bacterial isolates of Lactococcus garvieae, Aeromonas hydrophila, Streptococcus iniae, Streptococcus agalactiae and Yersinia ruckeri, respectively.CONCLUSIONS: This study shows that the spread of streptococcosis, lactococcosis, yersiniasis and Aeromonas septicemia and their frequent treatments has led to an increase in antibiotic resistance, especially against commonly used drugs such as erythromycin and oxytetracycline.

BACKGROUND: Heparin is a sulfated glycosaminoglycan. Blood is a common source for DNA detection in all kinds of samples, and anticoagulants such as heparin and ethylenediaminetetraacetic acid (EDTA) are used to prevent coagulation. Because heparin has a strong inhibitory effect on polymerase chain reaction (PCR), it is not used in samples that will be tracked by DNA. There are physical, chemical, and enzymatic methods to eliminate the inhibitory effect of heparin on PCR test.OBJECTIVES: First, to compare the intensity of the inhibitory effect of two anticoagulants, heparin, and EDTA, on the Real-Time PCR (qPCR), and then to investigate the impact of the heparinase enzyme present in the medium culture extract of Pseudomonas aeruginosa, on removing the inhibitory effect of heparin during the real-time PCR.METHODS: In the present study, two blood samples containing heparin and EDTA were subjected to a real-time PCR test to check the intensity of the inhibitory effect. Then, the medium culture extract of Pseudomonas aeruginosa was added to the heparinized blood sample infected with Escherichia coli bacteria in two groups with different conditions. In the first group, the DNA in the heparinized blood sample was extracted by the phenol-chloroform isoamyl alcohol method. Then, these samples were incubated with the extract of Pseudomonas aeruginosa bacteria culture medium at different hours, but in the second group, the samples were incubated at different hours before DNA extraction. Also, the DNA concentration in both groups was measured by a Nanodrop device, and finally, all samples were subjected to a real-time PCR test.RESULTS: The results of the research samples showed that although the heparinized blood sample contains more DNA concentration than the EDTA blood sample, it completely prevents genome replication. Also, incubating heparinized blood with Pseudomonas aeruginosa culture medium extract before DNA extraction for more than 24 hours removes the inhibitory effect of heparin during the real-time PCR, even at a lower cycle threshold than the EDTA-containing sample.CONCLUSIONS: The Pseudomonas aeruginosa culture medium extract may enable researchers to use heparinized blood samples for genome amplification and diagnosis without using expensive and limited commercial heparinase enzyme.

BACKGROUND: Clostridium perfringens (C. perfringens) is an anaerobic Gram-positive bacillus with spores, whose biotype A is responsible for a variety of diseases, including intestinal inflammation, bloody diarrhea, and gas gangrene, and hemorrhagic bowel syndrome. Genetic variety can explain the bacteria’s phenotypic diversity, geographic distribution, host specificity, pathogenicity, antibiotic resistance, and virulence. A molecular method using the pattern of DNA bands classifies bacteria based on the size of fragments produced by enzymatic digestion of the genome.OBJECTIVES: This study aims to standardize the polymerase chain reaction (PCR)- restriction fragment length polymorphism (RFLP) method in identifying the genetic diversity of C. perfringens biotype A isolates.METHODS: The genomic DNA of the investigated strains was extracted, and the complete sequence of the alpha toxin gene locus was synthesized using specific primers designed by PCR technique. Enzymatic cleavage of the synthesized amplicons was performed with the Mse l restriction enzyme, and the resulting fragments were separated by electrophoresis and analyzed by ImageJ and NTSYSPC software.RESULTS: The findings showed that the alpha toxin gene locus sequence may change and is not conserved. In this research, 4 different patterns were identified based on enzymatic cleavage. Mutations in this locus can lead to diversity in C. perfringens biotype A and the creation of new strains.CONCLUSIONS: The results of this research showed that the alpha toxin gene locus could be considered a DNA molecular marker in C. perfringens, and the PCR-RFLP technique can be used as a tool for typing this bacterium and estimating the phylogenetic relationships through comparative studies of nucleotide sequences.