Iranian Journal of Veterinary Research
Volume:16 Issue: 2, Spring 2015

The study was conducted to evaluate the effect of thermal stress on expression profile of genes related to apoptosis in peripartum Sahiwal cows. For this، twelve pregnant dry Sahiwal cows were selected from Livestock Research Centre at National Dairy Research Institute، Karnal. The cows were divided into two groups consisting of six Sahiwal cows each. Cows of group I calved during thermoneutral temperature conditions (THI=67. 3) and cows of group II calved in summer season (THI=79. 9). Blood sampleswere collected on -15، 0 and +15 days with respect to calving where day ‘0’ represents the day of calving. The peripheral blood mononuclear cells (PBMC) were separated and total RNA was isolated for the BCL-2 (B-Cell Lymphoma-2)، BAX (BCL-2 antagonist killer-1)، BAK (Bcl-2-associated X protein)، CASP-3 (cysteine-aspartic proteases-3) and P53 (tumour protien-53) mRNAs expression. It was found that there was up regulation of CASP-3 on the day of calving during both temperature conditions. Comparison between the two temperature conditions showed that expression of CASP-3، BCL-2، BAK، P53 and ratio of BAX/BCL-2 in PBMC increased during summer as compared to thermoneutral condition suggesting the susceptibility of these cells to apoptosis. Based on the above findings it can be concluded that during calving PBMC are more susceptible to apoptosis، and summer being more stressful potentiates the apoptosis of PBMC in Sahiwal cows.

Ichthyozoonotic Mycobacterium spp. poses health risks both to fish and humans. In this study، the presence of ichthyozoonotic Mycobacterium spp. was investigated in red mullet (Mullus barbatus barbatus) and surmullet (Mullus surmuletus)، widely caught species in the Mediterranean and the Aegean Sea. A total of 208 fish samples، provided from fishermen of Mersin province (Turkey) were studied. Using conventional methods، Mycobacterium spp. was isolated and identified at the genus level by PCR and at the species level by PCR-RFLP. Thirteen Mycobacterium spp. were detected in 13 (6. 25%) fish samples. Four mycobacteria were identified as M. genavense، three as M. fortuitum، three as M. scrofulaceum، one as M. marinum، one as M. vaccae and one as M. aurum. No signs of mycobacteriosis were observed in fish samples. Findings of this study can contribute to future studies of onichthyozoonotic Mycobacterium spp. in seafood.

Aflatoxins are secondary toxic metabolites produced by some Aspergillus spp. particularly، Aspergillus flavus and A. parasiticus that contaminate food and feed. The objective of this study was to evaluate the contamination of feedstuffs with Aspergillus spp. and detect genes involved in the aflatoxin biosynthesis pathway of A. flavus and A. parasiticus isolates. A total of 110 cow feed samples (comprised of silage، concentrate، hay and total mixed ration) from 30 industrial and semi-industrial dairy farms of Khorasan Razavi province، northeastern Iran، were examined using cultural and PCR methods. 68 (61. 82%) Aspergillus spp. were isolated from 110 samples of feedstuff. The predominant Aspergillus isolates were A. fumigates (21. 81%)، followed by A. flavus (17. 27%)، A. niger (10%)، A. parasiticus (8. 18%)، and A. oryzae (4. 54%). Fungal contamination levels of industrial and semi-industrial dairy farm samples were not significantly different (P>0. 05). Using four sets of primers، a quadruplex PCR was developed to detect genes (nor1، ver1، omtA and aflR) at different loci coding enzymes in the aflatoxin biosynthesis pathway of A. flavus and A. parasiticus strains. Out of 28 strains of A. flavus and A. parasiticus، 10 isolates (35. 71%) showed a quadruplet pattern indicating the important genes involved in the aflatoxin biosynthesis pathway، encoded for functional products. These isolates were confirmed to be aflatoxigenic by Thin Layer Chromatography. 18 isolates (64. 29%) had three، two and single molecular patterns. The results obtained by this study show that rapid and specific detection of aflatoxigenic molds is important to ensure the microbiological safety of feedstuffs.

An experiment was conducted to study the effects of an emulsifier (glycerol polyethylene glycol ricinoleate: GPGR) and different sources of fat on the performance of Khaki Campbell ducks. Ducks were assigned into five groups with three replicates (10 ducks/replicate) in each group. Treatments were a control diet (C1: without added oil and emulsifier)، control diet added with 2% soybean oil (C2). For the other three groups، maize was replaced with rice bran and added with 2% soybean oil plus emulsifier (T1)، 2% palm oil plus emulsifier (T2)، and 2% lard plus emulsifier (T3). Feed intakes were not affected (P>0. 1) by any dietary treatment. There were also no effects (P>0. 1) of dietary treatment on body weight gain and feed efficiency except for T3 group، where body weight gain was lower compared with other treatments، and feed efficiency was lower than C2، T1، and T2. The metabolizability of dry matter tended (P=0. 08) to decrease in T1، T2 and T3 groups than in C1 and C2 groups. Apparent metabolizable energy contents were significantly greater (P<0. 05) in the C2 group than in the C1 group، but were similar among C1، T1، T2 and T3 groups. Themetabolizability of fat and other nutrients were not affected (P>0. 10) by dietary treatments. Major carcass traits were unaffected (P>0. 10) among the treatments. In conclusion، soybean oil and palm oil with GPGR as emulsifier could be added in the diets containing high amount of rice bran without affecting the performance; whereas lard may adversely affect the performance of ducks.

This study was conducted to determine chromatin integrity and DNA damage by DNA electrophoresis and comet assays of buffalo fresh and frozen semen. Semen samples were collected from four buffalo bulls and evaluated after freezing for semen motility، viability، sperm abnormalities، chromatin integrity and DNA damage. A significant variation was found in semen parameters after thawing. Highly significant differences (P<0. 001) in chromatin integrity were observed between fresh and frozen semen. For the fresh semen، there was no significant difference between the bulls for chromatin integrity; however، a significant variation (P<0. 05) was detected in their frozen semen. No DNA fragmentation was observed by agarose gel electrophoresis. The percentage of sperm with damaged DNA detected by comet assay differed significantly between fresh and frozen semen. A significant negative correlation was recorded between motility and DNA damage (r=-0. 68، P<0. 05). Sperm abnormalities and DNA fragmentation were significantly positively correlated (r=0. 59، P<0. 05). In conclusion، DNA damage evaluation can provide reassurance about genomic normalcy and guide the development of improved methods of selecting spermatozoa with intact DNA to be used in artificial insemination.

The purpose of this experimental study was to evaluate the effect of chicken amniotic fluid (AF) on a cross section of rat sciatic nerves. Thirty adult male Sprague-Dawley rats weighing 275 to 300 g، were randomized into three groups treated with (1) amniotic fluid or AF (n=10)، (2) normal saline or NS (n=10)، and (3) sham surgery (n=10). The AF was aspirated from the amniotic cavity of incubating chick embryos at day 14. The sciatic nerve was exposed and sharply transected. Immediate epineurial repair was then performed. AF treated animals were given 2 ml/kg of the chick embryo AF subcutaneously، once daily، five times a week for up to 2 weeks. All animals were evaluated by sciatic functional index (SFI)، electrophysiology، histology، and immunohistochemistry at days 28 and 56 after surgery. The SFI difference between AF and NS groups at days 21 and 28 after operation was statistically significant (P<0. 05). The number of myelinated fibers in the AF group was significantly greater than that of the NS group at day 28 (P<0. 05). At days 28 and 56 after operation، the nerve conduction velocity (NCV) mean of the AF group was faster than that of the NS group، but the difference was not statistically significant (P>0. 05). The results of this study demonstrate that chick AF can enhance peripheral nerve regeneration.

Canine distemper virus (CDV) is the cause of canine distemper (CD) which is a severe and highly contagious disease in dogs. In the present study, a duplex reverse transcription polymerase chain reaction (RT-PCR) method was developed for the detection and differentiation of wild-type and vaccine strains of CDV. Four primers were designed to detect and discriminate the two viruses by generating 638- and 781-bp cDNA products, respectively. Furthermore, the duplex RT-PCR method was used to detect 67 field samples suspected of CD from Guangdong province in China. Results showed that, 33 samples were to be wild-type-like. The duplex RT-PCR method exhibited high specificity and sensitivity which could be used to effectively detect and differentiate wild-type and vaccine CDV, indicating its use for clinical detection and epidemiological surveillance.

Although Mycoplasma equigenitalium has been implicated in equine reproductive problems، its prevalence is largely unexplored due to the lack of specific diagnostic tests. To address this limitation، the authors developed and optimized species-specific primer pairs that target M. eguigenitalium rpoB (RNA polymerase B subunit) gene sequences. The specificity of the PCR assay developed in this study was determined using 12 field isolates including the type strain of M. equigenitalium and other Mycoplasma species. In the field study، a total of 122 mare and stallion samples comprising of 50 clinical and 72 random samples were subjected to speciesspecific PCR assay to detect M. equigenitalium in equine genital tracts. Mycoplasma equigenitalium (MEG) species-specific PCR detected 22. 13% positive samples; however، only 9. 01% of the samples were found to be positive using the conventional culture technique. The PCR established in this study could be used for rapid، specific and accurate diagnosis of M. equigenitalium strains. To the authors’ knowledge، this is the first report addressing the development and evaluation of species-specific PCR to detect M. equigenitalium.

Dilated cardiomyopathy (DCM) is accompanied by myocytes and connective tissue changes. Matrix metalloproteinases (MMPs) play important roles in cardiac remodeling. It seems that the gelatinases (MMP-2 and MMP-9) are effective enzymes in cardiomyopathy. Dilated cardiomyopathy was confirmed in 22 dogs (patient group) including 11 female and 11 male by clinical examination، auscultation، thoracic radiography and echocardiography. 17 healthy dogs (control group) with similar weight and breedto patients were also selected from referred cases to Small Animal Hospital of the Veterinary Faculty of Tehran University and the same diagnostic procedures were performed on them. After that، serum MMP-2 and MMP-9 of control and patient groups were measured by semi-quantitative zymography. Semiquantitative analysis of zymograms from canine serums with DCM showed that total MMP-9 in patients is more than control group، while there was no significant difference in total MMP-2 between the two groups. Pro-MMP-2 was not detected in patient group but its active form was present in both groups، of course MMP-2 activity inpatients was significantly more than control. Active form of MMP-9 was detected only in patients. Although pro-MMP-9 was present in both groups، its level in control group was significantly higher than patients. The heart enlargement was observed in the left، right or both parts. Statistically significant differences in active form of MMP-2 and MMP-9 levels were observed between different groups of heart enlargement (right، left and both parts) compared to control but this difference was not significant considering chambers affected and VHS (vertebral heart score) groups. In conclusion، although there are some changes in serum MMP-2 and MMP-9 levels in canine DCM، it seems that increase of MMP-9 is more prominent than MMP-2 and neither of them were affected by heart enlargement or VHS grade.

In the present study، the fertilizing potential of semen recovered from slaughtered bulls epididymis was evaluated after cryopreservation، by conventional techniques and flow cytometry methods. The cauda epididymal was dissected and sperm were recovered and evaluated for volume، sperm concentration، and membrane and acrosome integrity using a flow cytometer. Sperm fertility potential was tested by in vitro fertilization (IVF). For each bull، three trials of IVF were performed. Before freezing، on average، the sperm concentration was 216 ± 27. 5 × 106 sperm/ml. Sperm viability averaged 86. 5 ± 4%. The mean percentage of sperm with intact plasma membrane and acrosome before and after cryopreservation was 90. 7 ± 2. 9% and 90. 8 ± 1. 9% (P≥0. 05)، respectively. The fertilization rate using frozen/thawed epididymal semen averaged 64. 1 ± 3. 9% fertilization with no significant differences between bulls (P>0. 05). For the bull considered as control، the fertilization rate was 72. 2 ± 4. 5%، differing significantly (P>0. 05) from the frozen/thawed epididymal semen’s fertilization rate. In conclusion، it is possible to use in vitro techniques with cryopreserved spermatozoa obtained from bull’s epididymis using a controlled rate freezing method with a predetermined freezing curve، and with assessment of sperm’s viability by conventional techniques and flow cytometry methods، together with the fertilizing ability of cryopreserved epididymal spermatozoa.

A disease outbreak occurred in June 2012 among mud loach cultured on pond farms in Jangseong-gun، Jeollanam-do، Korea. Mortality rates reached up to 1. 2% in the farm per day. Typical clinical signs were bleeding ulcer at the middle portion of head and haemorrhagic erosion of the operculum. Based on biochemical characteristics، the causative bacterium isolated from diseased fish was identified as Aeromonas sobria. The isolate expressed two haemolytic genes، aerolysin (sob) and haemolysin (asa1) genes. Histopathologically، liver showed hepatocellular vacuolar degeneration and congestion in sinusoids. The spleen exhibited necrotized splenocytes and haemorrhagic pulps. In the kidney، glomerular destruction، renal tubular necrosis and haemorrhage were observed. Experimental infection (infectious dose of 106، 107، and 108 cfu fish-1) of healthy mud loach with the isolate resulted in the development of clinical signs similar to those seen in the farm. By injection with an infectious dose of 106 cfu fish-1، the mortality rate was 10. 3% within 7 days post infection. A mortality rate of 60. 9% was reached within 2 days when an infectious dose of 107 cfu fish-1 was used. Otherwise، all fish died within 1 day when injected with 108 cfu fish-1. The results demonstrated that A. sobria is involved in the morbidity and mortality of the farmed mud loach.

Equine piroplasmosis is a tickborne disease of equids with worldwide distribution، caused by Theileria equi and Babesia caballi. The aim of this study was molecular detection of T. equi and B. caballi in donkeys in northeastern Iran and investigate the association between positivity of piroplasm infection and host-related factors. In the present study، Blood samples were collected from 106 apparently healthy donkeys (Equus asinus) in North Khorasan province، Iran. Blood smears were prepared and stained by giemsa method. DNA was extracted from blood and then multiplex-PCR was done for detection of any piroplasms infection. According to the results، four donkeys showed T. equi in blood smears but B. caballi was not found. Also، fifty four donkeys (50. 94%) showed T. equi infection using multiplex-PCR. No siginificant difference was observed between the frequency of T. equi infection with hostrelated factors in donkeys. This is the first report on the molecular detection of eqiune piroplamosis in donkeys in Iran. Also، no significant association was found between the rate of T. equi infected animals.

The aim of this study was to reveal biometric peculiarities of New Zealand white rabbit antebrachium (radius and ulna) by means of three-dimensional (3D) reconstruction of multidetector computed tomography (MDCT) images. Under general anesthesia، the antebrachiums of a total of sixteen rabbits of both sexes were scanned with a general diagnostic MDCT. Biometric measurements of the reconstructed models from high resolution MDCT images were analyzed statistically. Consequently، when biometric measurement values of corresponding bones of antebrachium were compared، it was revealed that there was no statistical significance within both sexes but there were statistically important differences between both sexes in some biometric measurements. It has been suggested that the results from the study can shed light on future studies on the skeletal system and can form a modern point of view to anatomical education.

The objective of the present investigation was to determine the seroprevalence of Coxiella burnetii infection in free-range yaks in China. A total of 552 serum samples were collected from yaks in Gansu province، northwest China between April 2013 and January 2014، and antibodies against C. burnetii were evaluated using enzyme-linked immunosorbent assay (ELISA). Overall، 13. 59% (75/552، 95% CI: 10. 73-16. 45) of the examined animals were positive for C. burnetii antibodies. There was no significant difference in C. burnetii seroprevalence between female yaks (13. 78%، 95% CI: 10. 36-17. 19) and male yaks (13. 13%، 95% CI: 7. 89-18. 36) (P>0. 05). Coxiella burnetii seroprevalence in yaks in different age groups ranged from 10. 88% to 15. 26%، but the difference was not statistically significant (P>0. 05). Coxiella burnetii seroprevalence in yaks sampled in different seasons ranged from 12. 06% (autumn) to 18. 33% (summer)، but the difference was not statistically significant (P>0. 05). This is the first report of C. burnetii seroprevalence in free-range yaks in China، indicating the need for measures to be taken to control C. burnetii infection in free-range yaks in China.

The purpose of this study was to evaluate the effects of Scrophularia striata water extract on the quality and shelf life of the rainbow trout fillet during superchilled storage. Fish samples were treated with 1% and 3% S. striata water extract and then stored at -2°C for 20 days. The samples were analyzed periodically for chemical، microbial and sensory characteristics. Results indicated that incorporation of S. striata water extract on rainbow fillets caused the delay of lipid peroxidation and hydrolytic spoilage in 3% treated sample in comparison with the control sample at the last day of the experiment (P<0. 05). Moreover، fish fillets containing 3% S. striata water extract showed lower bacterial count than the control and 1% water extract supplemented samples (P<0. 05) during the experiment. According to sensory analysis results، 3% treated samples were acceptable even at the end of the 20-day storage. It was concluded that the effect of S. striata extract on fish samples was to retain their good quality characteristics and extend the shelf life during superchilled storage.

Veritable identification and differentiation of avian species is a vital step in conservative، taxonomic، forensic، legal and other ornithological interventions. Therefore، this study involved the application of molecular approach to identify some avian species i. e. Chicken (Gallus gallus)، Muskovy duck (Cairina moschata)، Japanese quail (Coturnix japonica)، Laughing dove (Streptopelia senegalensis)، and Rock pigeon (Columba livia). Genomic DNA was extracted from blood samples and partial sequence of themitochondrial cytochrome b gene (358 bp) was amplified and sequenced using universal primers. Sequences alignment and phylogenetic analyses were performed by CLC main workbench program. The obtained five sequences were deposited in GenBank and compared with those previously registered in GenBank. The similarity percentage was 88. 60% between Gallus gallus and Coturnix japonica and 80. 46% between Gallus gallus and Columba livia. The percentage of identity between the studied species and GenBank species ranged from 77. 20% (Columba oenas and Anas platyrhynchos) to 100% (Gallus gallus and Gallus sonneratii، Coturnix coturnix and Coturnix japonica، Meleagris gallopavo and Columba livia). Amplification of the partial sequence of mitochondrial cytochrome b gene proved to be practical for identification of an avian species unambiguously.

This case report illustrates the presence of diffuse idiopathic skeletal hyperostosis (DISH) in a fighting Bulldog. The dog was referred to the Veterinary Teaching Hospital، University of Agriculture Faisalabad Pakistan، with the presenting complaint of slowly progressing staggering gait، inability to stand on hind limbs and muscle stiffness in lumbo-sacral region. Hematological، serobiochemical and clinical examination were insignificant except presence of extensive new bone formation in the radiograph on the ventral of last 4 consecutive body lumbar vertebras (L4-L8) in lumbar region، running parallel to nuchal ligament. Diagnosis of DISH was made on the basis of clinical signs and radiographical examination which were suggestive of DISH. This report documents the first case of DISH in fighting Bulldog in Pakistan.

A dog was presented with history of progressive generalized subcutaneous emphysema and exercise intolerance secondary to dog fight. Radiographic evaluation confirmed extensive subcutaneous emphysema، pneumomediastinum and pneumoretroperitoneum. Surgical exploration revealed cricoid cartilage longitudinal fracture and cricotracheal detachment. The fractured cartilage was sutured and the cricoid cartilage and trachea was approximated using interrupted sutures. Concurrent cricoid cartilage fracture andcricotracheal detachment has not been reported in veterinary literature، which should be considered in any case of subcutaneous emphysema secondary to extrinsic laryngeal trauma.