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: Various flea species have already been reported from dogs, among which the most important ones include Ct. felis, Ct. canis, and P. irritans. Fleas can cause annoyance in dogs and human and transmit a variety of bacterial, fungal, and viral agents to the host. In addition, they could function as an intermediate host of Dipylidium caninum and Hymenolepis diminuta.  OBJECTIVES: Due to the lack of molecular species-associated identification data, we conducted the current study to differentiate Ct. felis and Ct. canis with molecular assay. METHODS: In the present study, 605 fleas were primarily collected from the dogs referred to Tehran Veterinary Faculty hospital. Subsequently, the flea species were identified under a microscope with morphological keys. Afterwards, COX1 genes of Ct. felis and Ct. canis were amplified via PCR and the locus was finally compared utilizing RFLP and sequencing. RESULTS: Totally, 605 fleas were isolated from 20 dogs. In morphological studies, three species were identified: Ctenocephalides felis, Ctenocephalides canis, Pulex irritans. Pulex irritans had the highest frequency (61.8 %). In molecular study, 552 bp fragment of COX1 gene in two species was amplified and seen on agarose gel. After sequencing, it was seen that two species sequences in COX1 locus had a similarity of 99 % and all of them depended on Ct. canis. In PCR-RFLP, in which Taq1 enzyme was used for differentiation of two species, the same result was obtained. CONCLUSIONS: Even though these two species of dog flea are distinct morphologically, their molecular differentiation using COX1 genes was not successful.

BACKGROUND: Full supply of the nutritional requirements of C57BL/6 mice plays an important role in preventing ulcerative dermatitis syndrome. OBJECTIVES: We conducted this work to asses ulcerative dermatitis in C57BL/6 mice colony and reduce it through dietary modification. METHODS: During a two-years period, a C57BL/6 mice colony in a laboratory animal breeding center was monitored for the presence of ulcerative dermatitis syndrome. In the first year, the animals were fed with outbred laboratory mice pellet and in the second year, the modified diet was used based on the nutritional requirements of this inbred mouse. RESULTS: The average numbers of litters per parturition were 3 and 6 in the first year and the second year, respectively. The litter's mortality rate in the first year was 75 % and zero in the second year. The total number of mice remaining in the colony was 220 in the first year, which increased to 1100 in the next year. Syndrome symptoms were observed in the adult mice in the first year. Alopecia was observed mainly in the back and neck and the severity of the symptoms was mild to severe. No mortality was observed in the mice, even in severe cases, and their recovery was spontaneous, yet very slow. In the necropsy, axillary lymphadenopathy, splenomegaly, and hepatomegaly were observed. In histopathological samples taken from the skin lesions, the accumulation of inflammatory cells and the spongiosis of cornea cells were observed. In lymph nodes, spleen, and liver, pathological changes were observed from the accumulation of inflammatory cells to necrosis. In the second year, the number of cases reduced significantly to 2 %. The severity of the symptoms was mild and their recovery was faster than that in the previous year. CONCLUSIONS: By fully supplying the nutritional requirements of C57BL/6 mice, we could significantly prevent the occurrence of ulcerative dermatitis syndrome.

BACKGROUND: Microbial growth and unfavorable taste and aroma are the leading reasons of meat spoilage under refrigerated temperatures. Today, plant essential oils are good alternatives to synthetic preservatives. In the meantime, nanotechnology has been instrumental in helping to apply the most effective plant compounds on the target cell. OBJECTIVES: The present study aimed to identify the compounds of essential oil of Cuminum cyminum and to investigate the microbial counts (total count, psychrotrophs, lactic acid bacteria, mold and yeast) after preparation of nanoemulsions of cumin essential oil via ultrasonic method and its nanoemulsions. Moreover, herein, we intended to evaluate organoleptic in chicken fillet samples under temperature of 4 °C. METHODS: The essential oil components were analyzed with GC / MS and the effects of Cuminum cyminum essential oil nanoemulsion were examined on microbial count and organoleptic properties in terms of color, odor, and overall acceptance (with 9-point hedonic scale) on Cuminum cyminum EO nanoemulsions in chicken fillets under 4 °C. RESULTS: The major identified components of Cuminum cyminum by extraction time were: beta- pinene (8.90 %), benzene- methyl (16.51 %), gamma- terpene (13.39 %), propanal (29.43 %), 1 -Isopropylidene-3-N-butyl2-cyclobutene (6.52 %), and benzene methanol (8.36 %). The results of microbial count at the last day of storage showed that the treatment containing 20 % and 25 % nanoemulsions significantly delayed microbial spoilage compared to other treatments (p < /em><0.05). The control treatment also indicated the highest level of microbial spoilage; as represented in Log 6, the maximum bacterial count in chicken fillets and the shelf life of BHT at 0.02 % concentration continued until day 3. Furthermore, in organoleptic evaluation, 20 % and 25 % nanoemulsion treatment scored higher than other treatments. CONCLUSIONS: Nanoemulsions of Cuminum cyminum EO at concentrations of 20 % and 25 % were able to delay the microbial spoilage of chicken fillets and produced organoleptically favorable results.

BACKGROUND: Magnetic cell targeting is a novel non-invasive cellular delivery technique. It improves stem cell delivery to and retention in the injury site. Labeling cells with superparamagnetic iron oxide nanoparticle (SPION) is one of the most important steps of this technique. Appropriate SPIONs selection is believed to be of vital importance. OBJECTIVES: The current study aimed to produce SPIONs which are capable of attaching to Mesenchymal stem cells surface (MSCs). METHODS: Dextran coated SPIONs were produced following co-precipitation method under N2 atmosphere. Bone marrow derived MSCs were isolated and cultured from rabbit humerus bone. Anti-rabbit CD44 monoclonal antibody was attached to the surface of SPIONs and MSCs and were labeled with this final product. SPIONs coating process, particle size, and antibody conjugation efficacy were evaluated using FT-IR, SEM, and Bradford protein measurement assay, respectively. Attachment of antibody-linked dextran coated SPIONs to MSCs was accessed utilizing Prussian blue staining, immunofluorescence analysis, and SEM analysis. RESULTS: Peaks of FT-IR at 3200 cm-1 and 2922 cm-1 are representative of dextran. The average particle size was 56.13±6.67. The average antibody-SPION conjugation ratio was 77.78±6.35%. The average percentage of the labeled cells in Prussian blue and IF analysis were 71.57±2.53 and 95.04±0.95, respectively. MSCs-SPIONs conjugation was also confirmed via SEM analysis. CONCLUSIONS: In conclusion, it could be inferred that mesenchymal stem cells could successfully be labeled with dextran coated-anti CD44 antibody conjugated- superparamagnetic Iron oxide nanoparticles. This product could be used for further in-vitro and in-vivo evaluations.

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: Vitamin C and Carum copticum with antioxidant activity may eliminate the negative effects of heat stress on broiler performance and immunity.OBJECTIVES: This experiment was conducted to investigate the effect of feeding heat-stressed broilers with Carum copticum seed powder (CSP) and vitamin C (VC) on their growth performance, blood metabolites, carcass characteristics, and meat quality.METHODS: Herein, we carried out A 2*3 factorial arrangement in a completely randomized design with six treatments, including three levels of CSP (0, 0.75 and 1.5 % of diet), two levels of VC (0 and 250 mg/kg of DM of diet), and four replicates, on 240 one-day-old male Ross 308 broilers. The collected data were analyzed using PROC GLM of SAS. Treatment means were compared employing the Tukey test.RESULTS: The effect of CSP on their performance was not significant whereas VC treatment improved weight gain significantly (P<0.01). Interaction of CSP and VC on weight gain and feed conversion ratio was significant as broilers fed with 250 mg VC and those fed with 1.5 % CSP had the best WG and FCR (P<0.01). The addition of VC decreased the concentration of alkaline phosphatase in the blood (P<0.05). The heterophil to lymphocyte ratio was reduced through the use of VC and 1.5 % CSP (P<0.05). CSP supplementation at the rate of 1.5 % increased the relative weight of bursa of fabricius (P<0.05). Dripping loss percentage was reduced in birds fed with 1.5 % CP and VC (P<0.01). The effect of CSP and their interaction on water holding capacity was significant and the highest amount belonged to the birds fed with 1.5 % CSP (P<0.01). Additionally, VC increased meat ash percentage compared to the control group (P<0.01).CONCLUSIONS: The results of this study indicated that addition of 250 mg VC and 1.5 % CSP could be conducive to improving growth performance, blood metabolites, and meat quality of broilers under heat stress conditions.

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: 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.