BACKGROUND: Bovine tuberculosis caused by Mycobacterium bovis is an important disease that has negative effects on public health and entails economic loss. Traditional controlling programs for cattle  focus on test and slaughter strategy, and false positive is one of the disadvantages associated with tuberculin skin test. To overcome this limitation, proteins with high specificity have to be utilized. OBJECTIVES: The objective of this study was to clone and express virulent protein CFP-10 from Mycobacterium bovis AN5. METHODS: Full-length genes of cfp-10 were amplified by PCR technique. In parallel, pET23a(+) and PCR products were double digested by EcoRI and HindIII. Ligation was performed at 16˚C followed by transformation into competence E. coli DH5α. After being identified with sequencing, the cloned vector was transformed into E. coli BL21. Induction was performed by isopropyl-β-D-thiogalactopyranoside (IPTG). Urea 8M was used to dissolve the expressed protein in the inclusion body form. Recombinant protein was purified by Nickle-Resin, and urea was eliminated by decreasing the gradient. RESULTS: The CFP-10 gene clone was proved by sequencing method. The CFP-10, as a 10 KDa protein, was confirmed by Western blotting using monoclonal antibodies. Based on the results, the recombinant protein was successfully cloned and expressed. CONCLUSIONS: The results showed that cfp-10 gene was successfully cloned and expressed in prokaryotic system, indicating that this recombinant protein could be utilized in diagnostic kits against bovine tuberculosis in the future.

BACKGROUND: Cystic echinococosis (CE) is known to be one of the most important zoonotic diseases in different parts of Iran. Even though it causes major health problems, there is limited information regarding its transmission cycles and strain of this infection in eastern Iran. OBJECTIVES: The present study aimed to characterize the strain of Echinococcus granulosus cysts in the slaughtered sheep and goats in Birjand area using morphological and molecular criteria. METHODS: Isolates of E. granulosus were collected from sheep (30) and goats (30) from Birjand slaughterhouse and characterized employing both DNA (PCR-RFLP of ITS1) and morphological criteria (metacestode rostellar hook dimensions). In addition, the fragments of the genes coding for ITS-1 were sequenced. RESULTS: Among the two different identified strains/genotypes (sheep and camel), the sheep strain appeared to be the most common genotype of E. granulosus affecting sheep and goats. All of the 30 sheep samples and 20 out of 30 goat samples were infected with sheep strain. However, the camel genotype was only observed in the goats and 10 out of 30 goat isolates were infected with the camel genotype. The camel genotypes had RFLP patterns, which were different from the RFLP patterns of the rest of isolates (sheep strain). CONCLUSIONS: The results of this study revealed that the ‘sheep’ strain was the most prevalent strain in sheep and goats in this area. Moreover, the camel genotype (G6) was confirmed to trigger infection in the slaughtered goats of Birjand area.

BACKGROUND: Oxidative stress is involved in pathogenesis of Polycystic Ovary Syndrome (PCOS). Lutein is a herbal compounds with antioxidant properties. OBJECTIVES: The current research aimed to evaluate the effect of lutein on body weight and histomorphometry of uterus in experimental PCOS induced with Dehydroepiandrosterone (DHEA) in mouse models. METHODS: Twenty-four female NMRI mice aged 20 days and weighing 14-17 g were randomly assigned to four equal groups: control, experimental PCOS, and PCOS treated groups with 125 and 250 mg/kg lutein. The induction period of PCOS with oral administration of DHEA (6 mg/100 g, daily) was 21 days and lutein treatment was followed by the induction period of 28 days. The mean body weight of the groups was evaluated on day 0, day 21 (at the end of DHEA treatment), and day 49 (at the end of treatment period) with lutein. The mean diameter of the uterine wall, the mean overall thickness of the uterine wall, the average thickness of the endometrium, myometrium and uterine epithelium, along with the number of endometrial gland branches were measured utilizing light microscope. RESULTS: The results revealed that body weight in the PCOS group was significantly higher than that in the control group on days 21 and 49. Treatment with 125 and 250 mg/kg of lutein reduced body weight in the lutein treated groups compared with PCOS (p < /em><0.01). The mean uterine wall diameter, mean total uterine wall thickness, mean thickness of endometrium, myometrium, and uterine epithelium with the number of uterine endometrial branching were significantly lower in the control and lutein treated groups compared to those in the PCOS group (p < /em><0.05). The use of both doses of lutein (125 and 250 mg / kg) significantly improved uterine histopathological indices, particularly the mean uterine wall diameter (p < /em>=0.0001) compared to the PCOS group. CONCLUSIONS: Lutein could improve the side effects of induced PCOS by DHEA on body weight and uterine parameters.

BACKGROUND: Several studies have reported the application of mesenchymal stem cells (MSCs) for the treatment of equine musculoskeletal disorders. Bone marrow (BM) and adipose tissue (AT) are the main sources for isolation of MSCs. Although the MSCs derived from various sources have similarities, certain differences have been reported in terms of biological, proliferative, immunological, and differentiative characteristics. They are also different concerning gene expression profiles. So, comparative gene expression analysis on mRNA level via quantitative real-time polymerase chain reaction (qRT-PCR) is believed to be of great value for better characterization of MSCs from different sources.

BACKGROUND: Sex determination is an essential issue in certain fish species, like sturgeons. During the rearing management, distinguishing between male and female fish is an important process for producing meat or caviar.OBJECTIVES: The current study aimed to find the feasibility of electric organ discharge (EOD) frequency ‎generating to introduce a novel method of sex determination in three freshwater species, namely the ‎black ghost knife fish (Aptronotus albifrons), goldfish (Carassius auratus), and the Sterlet ‎sturgeon (Acipenser ruthenus), as non-electric fishes.METHODS: In this regard, following sex detection, EOD frequency ‎of fishes were recorded via an oscilloscope. In addition, knife fish was used as a reference sample on which a lot of work has been done to evaluate the accuracy of the method and the use of the oscilloscope. Furthermore, the EOD frequency of male and female ‎Sterlet sturgeons was recorded and reported for the first time in this study.RESULTS: The frequency ‎magnitude was significantly different in genders of Sterlet (P<0.05) with higher EOD frequency in males ‎than in females. There was no recorded frequency in goldfish ‎male and female. EOD frequencies, which were recorded in male and female of black ghost fish, ‎represented significant differences compared with the other studies (P<0.05).CONCLUSIONS: The results achieved ‎in this study revealed that sex differences are one of the effective parameters in terms of EOD ‎frequency inequality; this non-invasive method can be utilized for sex identification of valuable ‎species, like sturgeon. However, it is required to gather advanced equipment and conduct further ‎research in physiology and biology of reproduction for accurate recording of electrical frequencies ‎and detection of their origins.

BACKGROUND: Serological survey of NDV infection from LBMs would give a good picture of Newcastle disease ecology in a country.OBJECTIVES: This cross-sectional study was carried out to evaluate the seroprevalence of Newcastle disease in live bird markets, bird parks, and zoos in Iran.METHODS: From July to December 2016, blood samples were collected from different bird species in each unit. The serum samples were evaluated via Hemagglutination inhibition test. The seropositive sample was considered a serum with antibody titer 4 or more (1.16) in HI, and units with at least one seropositive bird were considered as the contaminated unit. In this study, the serum samples were taken from various bird species (N=2292) selected from 127 bird markets, bird parks, and zoos distributed in 22 Iranian provinces.RESULTS: Among the 127 sampled units, 70 (55.12 %) were found to be seropositive. In addition, among the 2292 sampled birds, the number of seropositive birds were found to be 495. Among different bird species, the highest sero-positive prevalence belonged to chickens and turkeys with 38.7 % and 32.89 %, respectively.CONCLUSIONS: The results of this study suggested a high prevalence of Newcastle disease in the live bird markets, bird parks, and consequently, across our country. Given the importance of this infectious disease, it is essential to apply appropriate controlling measures, including continuous surveillances of circulating viruses and vaccination programs with conventional vaccines, such as heat-resistant vaccines. On account of the important role of rural poultry and wild birds in Newcastle disease distribution, controlling the disease in rural poultry and continuous surveillance in both can prevent the spread of NDV, particularly to the commercial poultry.

BACKGROUND: Leucine is one of the subgroups of amino acids, which play an important role in the anabolism of muscles, adipose tissue, and the liver by stimulating insulin secretion.OBJECTIVES: Effects of different levels of leucine were investigated on carcass yield, characteristics, and quality, and expression of insulin-like growth factor (IGF-1) and insulin genes in male broilers.METHODS: Five levels of L-leucine (100, 110, 120, 130, and 140 % of Ross strain requirements) were tested with 250 male one-day-old chicks in a completely randomized design with five treatments and five replicates (containing 10 chicks each). On day 42 of their age, the blood samples of two birds from each replicate (10 birds per treatment) were taken to determine serum IGF-1 gene expression. Subsequently, these birds were slaughtered for analysis of carcass characteristics and quality, and collecting the samples of liver and breast for expression of IGF-1 and insulin genes.RESULTS: Body weight increased by consumption of 140 % of leucine as compared to 100 %. Reduction in feed conversion ratio was observed by feeding 140 % of leucine level. The IGF-1 gene expression of breast and liver increased by 110 % of leucine level. Moreover, feeding 110 % of leucine level caused a higher expression of insulin gene in breast and liver. Consumption of 130 % of leucine improved the meat protein, fat, and ash contents. Furthermore, consumption of 110 % of leucine increased the serum IGF-1 concentration.CONCLUSIONS: Consumption of leucine in broiler diets was found to increase the expression of IGF-1 and insulin genes and consequently, improve the performance and carcass quality. It also decreased the abdominal fat in broiler chickens.

BACKGROUND: The veterinary Rabies vaccine was produced using BHK-21 cells and PV strain. Although various protocols have been suggested for virus purification, they have an adverse effect on the final production and require further optimization.OBJECTIVES: The present study aimed to optimize the concentration and purification of the virus for rabies vaccine production.METHODS: First of all, the Pasteur virus strain (PV) was cultured by using BHK 21 cells with DMEM media contain bovine fetal serum (7 %) for five days. Subsequently, the virus purification was done via tangential flow filtration (TFF) system. The quality of purifying viruses was an assessment with titration and SDS-PAGE. Secondly, the virus inactivation was optimized using Minitab software based on three factors, namely time, temperature, and concentration. Afterwards, the inactivity of the samples was tested on mice. Finally, the virus potency was evaluated by the National Institute of Health (NIH) method.RESULTS: The viral titration test in TFF samples revealed that viral titer increased in comparison with the control group (P<0.05). The SDS-PAGE analysis of the purified and concentrated samples showed that the purified virus via TFF had a higher purity compared to the not-concentrated samples. Moreover, the NIH test indicated a 10-fold increase in potency result in the TFF group.CONCLUSIONS: The present study implied that the TFF method is highly suitable for condensation and purification of a high volume of viral fluid and could be applied on an industrial scale to increase the potency of the vaccine produced.

BACKGROUND: As the population grows, there is further need to food, and fish is not an exception. Several fish species are potential sources of common parasites between humans and fish. One of the important diseases common between human and fish is Anisakiasis. The parasitic agent of this disease is the larval stage of the Anisakid family nematodes, including Pseudoterranova and Anisakis.OBJECTIVES: In this study, Epinephelus fish, one of the edible and commercial valuable fishes of the Oman Sea, was examined for the presence of nematodes of the Anizakidae family.METHODS: Fifteen out of the 26 specimens were infected with Nematodes. Nematodes were isolated from fish abdominal area. For morphological study, each nematode sample was first clarified with lactophenol. Subsequently, it was examined using an optical microscope. After morphological examination of these nematodes, DNA extraction was performed. Using primers related to a part of cytochrome oxidase subunit 1 (Cox1), PCR products were 710 bp for PCR reaction. Finally, the amplified fragment was sequenced.RESULTS: The larvae were about 1 to 3 cm long, white, and often twisted. At the anterior end of the parasite, a button was seen, and in some larvae, a terminal spine was observed. In certain larvae, a small abdomen at the end of the esophagus can be seen. Out of the obtained 26 nematode specimens, eight Anisakis specimens were identified following morphological analysis. These specimens had terminal spines and three anterior lips. After sequencing, Pseudoterranova nematode was identified to belong to aniakidae family. Separate clad tree showed paraphylitic for isolated Pseudoterranova.CONCLUSIONS: Morphological examination categorized isolated larvae as the Anizakidae family. Other molecular results of this nematode showed Pseudoterranova krabbei. The results of sequencing this parasite were recorded in the gene bank under the Accession number: MK317965. This nematode was initially isolated from the Oman Sea Epinephelus fish.

BACKGROUND: Open wounds affecting the distal part of limbs are commonly seen in horses. Due to certain factors, such as limited connective tissue available, potentiated growth of excessive granulation tissue, risk of contamination, and poor response to common treatments, healing of these wounds becomes a major problem for veterinarians on a number of occasions. Application of Mesenchymal Stem Cells (MSC) for enhancing wound healing has received a great deal of scientific attention. Among the MSCs, those derived from adipose tissue are frequently used owing to their availability, large number of cells after the primary harvest, and the capacity to differentiate to different cell lines.OBJECTIVES: The current study aimed to evaluate type 1 and 3 collagen genes expression in horse distal limb wounds treated via adipose-derived Mesenchymal Stem Cells and its comparison with bone marrow-derived stem cells.METHODS: After treatment of the experimental open wounds created in the distal limbs of four horses via autologous MSCs, real-time PCR was used for evaluating and comparing the expression of type I and III collagen genes in the healing wounds.RESULTS: Significant differences in the expression of type I and III collagen genes were observed between the treatment groups. Despite the fact that the greatest collagen genes expression belonged to bone marrow-derived MSCs, no significant differences were seen with adipose-derived MSCs.CONCLUSIONS: Owing to the advantages and an acceptable performance, adipose-derived MSCs could be considered as a novel approach to enhancing limb wound healing in horses.