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: Toxocaiasis is an important zoonotic disease caused by the second stage larvae of Toxocara canis and Toxocara cati. Toxocariasis is the most common worm infection in several temperate countries and causes severe complications. C-type lectin is one of the larval stage products of this parasite. It is involved in immune responses, including cellular signals in vertebrate immunity, activation of innate immunity in vertebrates and non-vertebrates, and induction of homeostasis.  OBJECTIVES: The current research aimed to optimize the production of recombinant C-type lectin of Toxocara canis and investigate its antigenic properties. METHODS: Reference nucleotide sequence of lectin type C (CTL) of Toxocara canis (T. canis) was extracted from Genbank. Recombinant Plasmid, pET32a, including the desired sequence was then constructed by GENERAY. The recombinant plasmid was transformed to Escherichia coli strain BL21 (DE3). The expression of the recombinant protein was investigated using SDS-PAGE and dot blot methods and approved with human T. canis positive serum. RESULTS: The findings of the present study showed that optimization and high level production of recombinant protein expression was achieved by selecting Escherichia coli (BL21 (DE3), 37 °C temperature for 4 hours after induction and 1 mM IPTG concentration. After optimization, the recombinant protein was obtained at a concentration of 1160±0.6 µg/mL. The molecular weight of the resulting recombinant protein was 42 kDa. Recombinant plasmid passage in Escherichia coli DH5α strain caused a significant increase in recombinant protein expression. The results of condition optimization evaluation, with SDS-PAGE and Dot blot methods, showed that the highest production of recombinant C type lectin protein of T. canis was obtained under optimized conditions. CONCLUSIONS: Based on these results, to produce reasonable amounts of specific C-type lectin recombinant protein, further studies are needed to evaluate its immunogenicity and protection against Toxocara canis infection.

BACKGROUND: Medicinal plants have a long history in the treatment of parasitic diseases and are usually free of side effects. Removing and killing tomonts and theronts of this parasite can prevent the parasite pathogenicity in fish. OBJECTIVES: The present study aimed to investigate the inhibitory effect of ethanolic extract of Zataria multiflora on both free and parasitic life stages. METHODS: Three soluble forms of Zataria multiflora extract (80 ml/L) were prepared, which contained freshly prepared solution, stored solution at room temperature for one month and stored solution at refrigerator for one month. Afterwards, different dilutions were prepared for each of the solutions and the tomonts and Theronts were treated with different dilutions of the extract. RESULTS: Zataria multiflora extract can kill all theronts at concentrations of 20 ml/L during 6.04 to 6.37 min and concentration of 10 ml/L during 31/2 to 32.4 min, respectively. However, at these concentrations, only 3.46-6.11 % of tomonts were killed herein after 4 hours. CONCLUSIONS: Tomonts were found to be more resistant to the parasiticides than theronts. The use of Zataria multiflora extract significantly reduced tomont reproduction and decreased Ich infectivity prevalence and intensity. Zataria multiflora extract prevented Ich infestation in naive fish and effectively treated infected fish.

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: Mastitis could be detected in various ways, including physical, on-farm, and laboratory tests. OBJECTIVES: The present research aimed to evaluate the accuracy of the diagnosis of mastitis using lactate dehydrogenase (LDH)-based dipsticks and to assess these dipsticks with regard to the effects of different lactation days, the amount of milk production, and parity. RESULTS: Considering bacteriologic culture as a gold standard method for the diagnosis of subclinical mastitis, the sensitivity and specificity of LDH test were 68.9 % and 54 %, respectively. The results revealed a high correlation between SCC and LDH. A statistically significant relationship was observed between the response of dipstick and CMT results; with the increase in the CMT score, the score of LDH dipstick increased. By investigating the effects of lactation days, the amount of milk production, and parity, it was determined that the chance of having subclinical mastitis in cows with positive dipstick result was 5.59 times greater than that in cows with negative dipstick result. There were no significant relationships among SCC, LDH, and CMT with lactation days and milk production; meanwhile, with the increase in parity, the three above-mentioned variables showed significant increase. CONCLUSIONS: The results of the present study indicated that the best methods for subclinical mastitis detection were SCC, CMT, and LDH based dipsticks, respectively.

BACKGROUND: Polycystic kidney disease is the most prevalent inherited genetic disease in Persian cats, which is caused by mutations in PKD1 and PKD2 genes. Due to the accumulation of fluids inside the cysts and their pressure on the renal parenchym, the patient is prone to developing symptoms of chronic renal failure.OBJECTIVES: The present study aimed to compare ultrasonography and molecular tests in diagnosis of autosomal dominant polycystic kidney disease.METHODS: This study was performed on 97 Persian cats, including 46 male and 51 female cats, with an average age of 6 years (minimum 2 months and maximum 14 years). All the cats were evaluated for the presence of disease using ultrasound and molecular methods.RESULTS: Among 97 females, 32 (33 %) were found to be positive for PKD on the basis of presence of anechoic cysts. In molecular tests, all the cases with cysts in the ultrasonography had mutation in PKD1 gene and 13 cases (13 %) without cysts in ultrasonography were diagnosed to be positive through molecular technique. Among 97 studied cats, 45 (46 %) showed mutated genes. The degree of agreement between the two methods of ultrasonography and PCR was determined by calculating Kapa 0.725 (Cl: 0.592-0.895). The sensitivity and specificity of the ultrasonography were calculated to be 77.11 % and 100 %, respectively.CONCLUSIONS: Imaging and molecular methods were utilized to diagnose the disease. The more frequent use of the molecular methods for the diagnosis of the disease compared to the use of ultrasound could be attributed to the higher sensitivity of the molecular technique, the small size of the cysts, the low number of cysts, the low age of the animal, and the presence of cysts in the medula of the kidney. Therefore, the molecular method could be recommended for screening the disease in the early stages. It can also be employed in breeding programs and the removal of cats with this mutated gene.

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: Optimal levels of minerals in the diet are crucial for improving performance and immune response against the environmental stresses.OBJECTIVES: This study was conducted to evaluate the effects of water supplementation of chelated trace mineral (CTM) on growth performance, hematology, blood biochemistry, antioxidant status, and antibody response in broilers reared under heat stress condition.METHODS: A total of 240 male Ross 308 broiler chickens were randomly allocated into one of the following five treatments (each consisting of four replicates): (1) positive control (PC; thermoneutral condition), (2) negative control (NC; heat stress condition), (3) low-CTM (NC + low CTM level in tap water), (4) medium-CTM (NC + medium CTM level in tap water),  and (4) high-CTM (NC + high CTM level in tap water). The CTM levels in the tap water for the low-CTM treatment were 0.1, 0.15, 0.3, 0.4, and 0.45 mg/bird/day on days 1-10, 11-20, 21-30, 31-40, and 41-50, respectively. The respective values were 1.0, 1.5, 3.0, 4.0, and 4.5 for the medium-CTM, and 10, 15, 30, 40, and 45 mg/bird/day for the high-CTM water.RESULTS: The average weight gain during the whole experimental period in the medium-CTM treatment was higher compared to that in the negative control treatment, but lower than that in the positive control treatment (P<0.05). The heterophil/lymphocyte ratio and total antibody titers against Newcastle disease in the medium-CTM and high-CTM groups were similar to those of the broilers in the PC group, yet better (P<0.05) those of the NC group. The serum superoxide dismutase activity was also higher (P<0.05) in the high-CTM group compared to that of the NC group.CONCLUSIONS: The results suggested that the water supplementation of CTM at a moderate level (1.0, 1.5, 3.0, 4.0, and 4.5 mg/bird/day during different rearing periods) could be a strategy to ameliorate the detrimental effects of heat stress in broilers.