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Veterinary Research Forum - Volume:7 Issue: 3, Summer 2016

Veterinary Research Forum
Volume:7 Issue: 3, Summer 2016

  • تاریخ انتشار: 1395/06/30
  • تعداد عناوین: 14
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  • Kolsum Choobineh, Saeed Zavareh*, Mojdeh Salehnia, Mohammad Taghi Ghorbanian, Seyed Hassan Paylakhi Pages 181-188
    It was assumed that uterine stem cells are responsible for the unique regenerative capacity of uterine. Therefore, the aim of the present study was to investigate the expression of the pluripotent stem cell markers in the mice uterine tissue during different stages of estrous cycles. Twelve virgin female NMRI mice (6 to 8 weeks old) were considered at proestrus, estrus, metestrus and diestrus according to the cell types observed in the vaginal smear and underwent hysterectomy operation. Quantitative real-time polymerase chain reaction (PCR) and immunohistochemical staining for pluripotent stem cell markers (SOX2, OCT4, KLF4, and NANOG) were performed. Immunofluorescence staining revealed that expression and localization of the pluripotency markers SOX2, OCT4, KLF4, and NANOG at the protein level were not different throughout estrous cycle. Also, mRNA of pluripotency markers was detected in all tested samples. However, there were no significant differences in their genes expression at each stage and during the estrous cycle. Different hormonal profile during the estrous cycle could not affect expression of pluripotent stem cell markers in uterine tissue.
    Keywords: Estrous Cycle, Mice, Pluripotent Stem Cells, Uterine Tissue
  • Nasibeh Mohammadagheri, Ramin Najafi *, Gholamreza Najafi Pages 189-195
    The present experiment was conducted to evaluate the effects of organic acids and phytase enzyme supplementation on performance and intestinal histomorphology of broilers. The experiment was done in a factorial arrangement 2 × 2 × 2 based on completely randomized design with eight treatments, five replicates with 12 chicks in each until 42 days of age. Diets included natural vinegar (0 and 2%), citric acid (CA; 0.00 and 1.00%) and phytase enzyme (PHY; 0.00 and 500 FTU phytase per kg of feed). One bird from each treatment replicate was randomly selected and slaughtered to evaluate the small intestinal morphology on 42 days of age. Analysis of results showed that vinegar increased feed consumption and body weight gain in total experimental period (p 0.05), while CA significantly decreased feed consumption on 0-14 days of age (p 0.05). No effect was observed on performance in interaction of organic acids together and with PHY group (p > 0.05). In duodenum CA increased the villusheight and width (p 0.05) and PHY enzyme increased villuswidth (p 0.05) and decreased crypt depth (p 0.05). On the other hand, CA along with PHY significantly decreased crypt depth (p 0.05). In jejunum PHY alone and in combination with vinegar increased the goblet cells numbers (p 0.05), whereas vinegar significantly increased the goblet cells numbers in ileum (p 0.05). The muscular thickness in duodenum, jejunum, and ileum was not affected among different treatment groups. The results showed that supplementation of organic acids and phytase together in this experiment, with no negative effects on each other, improved their effects on some parameters.
    Keywords: Broiler chicken, Intestinal histomorphology, Organic acids, Phytase enzyme
  • Abolfazl Ghaniei* Pages 197-202
    Mycoplasma synoviae (MS) is a pathogen responsible for respiratory and locomotor disorders and causes major economic losses in poultry industry. Early and accurate diagnosis of MS infection plays a major role in control of the infection. This study was conducted to characterize Iranian field isolates of MS isolated from broiler chickens of West Azarbaijan province (Northwest of Iran), and differentiate them from vaccine strain MS-H. Two encoding genes, 16S rRNA and vlhA were employed. PCR results using primers related to 16s rRNA and vlhA genes were analyzed and compared. Out of 21 field samples, eight samples (38.0%) were positive using both sets of primers. Amplified products of vlhA gene were sequenced for MS strain identification. The results showed that Iranian field isolates of MS had high nucleotide and amino acid similarity. Iranian field isolates were distinct from vaccine strain MS-H. Results presented in this study showed that characterization of field isolates of MS by sequencing of vlhA gene and is beneficial for strain typing and differentiating them from vaccine strain. To our knowledge, this is the first study characterizing vlhA gene of MS isolates from broiler chickens in the West Azarbaijan province.
    Keywords: Broilers, Iran, Mycoplasma synoviae, vlhA
  • Afrooz Sharifi, Morteza Chaji *, Tahereh Mohammadabadi Pages 203-211
    The aim of this study was to evaluate the effects of adding sulfuric acid to sugarcane topssilage on rumen bacteria and whole rumen microorganisms (WRM) and compare the digestibility of sugarcane tops treated with different amount of urea, molasses and sulfuric acid between Holstein cow and Khouzestan buffalo. Regardless of the type of the treatment, potential of gas production (B) by cow WRM (130.670 mL) was more than buffalo (104.060 mL) (p
    Keywords: Bacteria, Buffalo, Molasses, Sulfuric acid, Urea
  • Yasser Shahbazi* Pages 213-219
    Doogh is the most popular and commonly consumed yoghurt-based Iranian drink. The aim of this study was to investigate the antibacterial effects of Ziziphora clinopodioides essential oil (ZEO) at 0.10 and 0.20% concentrations, nisin at 250 and 500 IU mL-1, and their combination against Salmonella typhimurium and Staphylococcusaureus in doogh during storage at 4 ˚C for 9 days. Nine batches were studied as follows: control: no ZEO or nisin added, A: 0.10% ZEO, B: 0.20% ZEO, C: 250 IU mL-1 nisin, D: 500 IU mL-1 nisin, E: 0.10% ZEO 250 IU mL-1 nisin, F: 0.10% ZEO 500 IU mL-1 nisin, G: 0.20% ZEO 250 IU mL-1 nisin and H: 0.20% ZEO 500 IU mL-1 nisin. Based on gas-chromatography and mass spectrometry, carvacrol (65.22%), thymol (19.51%), p-cymene (4.86%) and ɣ-terpinene (4.63%) were the major components of ZEO. The populations of S. typhimurium and S.aureus in samples treated with all concentrations of the ZEO and nisin were kept below 1 log CFU mL-1 on day 5 of storage, while the count of S. typhimurium and S.aureus was found as 2.72 ± 0.02 and 2.21 ± 0.00 log CFU mL-1 on day 5 for untreated samples, respectively. The ZEO separately and in combination with nisin, was very effective against these two common food-borne pathogens. The ZEO alone and in combination with nisin could be considered as a potential strong antimicrobial agent that can be used for the growth inhibition of aforementioned bacteria in food products especially doogh.
    Keywords: Doogh, Nisin, Salmonella typhimurium, Staphylococcus aureus, Ziziphora clinopodioides
  • Mohammad Ali Ebrahimi Saadatlou*, Rasoul Shahrouz Pages 221-226
    In the present study, the lenses and ciliary bodies of 20 ostrich eyes were studied macroscopically and microscopically. The histological slides were studied after staining by hematoxylin and eosin, Verhoeff, Van Gieson, and periodic acid-Schiff (PAS). Posterior surface of lens was more convex than its anterior surface. The average lens diameter and thickness were respectively measured as 1.43 ± 0.00 and 0.85 ± 0.00 cm. The average ciliary body thickness was measured as 1.48 ± 0.01 cm. In addition, the ciliary body was seen annular with mean horizontal and vertical external diameters as 4.80 ± 0.07 and 4.36 ± 0.06 cm, respectively. The retina is extended on ciliary body in this bird. The number of ciliary body processes was about 120. The epithelium of lens was cuboidal and the lens capsule had intense positive PAS reaction. Also, the anterior surface of capsule was thicker than its posterior surface. The lens fibers in the central part were thicker than other parts. Elastic and collagen fibers were not observed in the lens. The epithelium of ciliary processes had two layers; superficial cuboidal non-pigmented layer, and deep heavily pigmented layer. The ciliary body was supported by a hyaline cartilage. In addition to the smooth muscle fibers, many isolated skeletal muscle fibers were also seen in ciliary body. In conclusion, the lens and ciliary body of ostrich were similar to other birds, although there were little differences in anatomical dimensions and histological characteristics.
    Keywords: Ciliary body, Histoanatomy, Lens, Ostrich
  • Ramezan Ali Jafari *, Hossein Motamedi, Elham Maleki, Reza Ghanbarpour, Mansoor Mayahi Pages 227-233
    This study was conducted to reveal the phylogenetic background, to detect the genes encoding TEM, SHV and CTX-M-15 extended-spectrum β-lactamases (ESBL), and to analyze their distribution in phylo-groups of 150 Escherichia coli isolates from broiler chickens in Ahvaz (Southwest of Iran). Seventy- five cloacal swabs from healthy birds (fecal isolates), and 75 heart blood samples from birds with colibacillosis (septicemic isolates) were obtained. All isolates were phylotyped and screened for ESBL genes by polymerase chain reaction (PCR). The fecal isolates belonged to four main phylo-groups, including 41 isolates (54.67%) to A, 9 (12.00%) to B1, 5 (6.67%) to B2, and 20 (26.67%) to D. Of septicemic isolates, 37 isolates (49.33%) were classified as phylotype A, 5 (6.67%) as B1, 10 (13.33%) as B2, and 23 (30.67%) as D. In molecular analysis, a total of 72 isolates (35 fecal and 37 septicemic) were identified to harbor ESBL genes, which were distributed in phylo-groups A, B1, B2, and D. Regardless of the type of isolate, blaCTX-M-15 gene was the most common genotype, followed by blaTEM and blaSHV genes. This study suggests that broiler chickens in Iran are infected to ESBL genes- harboring Escherichia coli strains which may be spread to the food chain through fecal contamination of carcasses during slaughtering.
    Keywords: Beta, lactamase, Chicken, Escherichia coli, Phylogeny
  • Sahar Nouri Gharajelar*, Rajab Ali Sadrkhanloo, Masoud Onsori, Adel Saberivand Pages 235-239
    Cryopreservation has the capacity to extend spermatozoa’s lifespan and viability. In addition, the semen samples can be collected, preserved and stored or sent to distant locations and still be used long after the death of the semen donor. In this study for the vitrification of dog sperm (fresh and swum-up sperm), different cryopreservation mediums on the basis of glycerol, milk and egg yolk were used. Then, all of the samples were vitrified in the liquid nitrogen and thawed at least 48 hr later for re-examination of sperm parameters. The sperm parameters before and after cryopreservation in all groups were compared. It was found that during vitrification process, spermatozoa were damaged by the mechanical blows in centrifugation during swim-up processing, so they had less resistance than fresh semen. The examination of different cryoprotectants revealed that milk has better effects on the cryopreservation of semen than glycerol and egg yolk. With the comparison of the effects of glycerol and egg yolk as cryoprotectants, it was found that glycerol had better effects than egg yolk on the cryopreservation of the semen. In conclusion, milk might be used as a cryoprotectant instead of glycerol for canine sperm cryopreservation.
    Keywords: Cryoprotectant, Dog, Sperm, Vitrification
  • Rahem Khoshbakht*, Mohammad Tabatabaei, Saeid Hoseinzadeh, Mojtaba Raeisi, Hesamaddin Shirzad Aski, Enayat Berizi Pages 241-246
    Althoughpoultry meat is considered as the main source for human Campylobacter infections,there is limited information about non-poultry sources. The present study was aimed to investigate the prevalence and the antibiotic resistance of thermophilic Campylobacter spp. in fecal samples of the cattle and sheep in Shiraz, Iran. A total of 302fecal samples were obtained from clinically healthy, slaughtered cattle and sheep from Shiraz slaughterhouse. The animals were clinically healthy before being slaughtered. The samples were cultured according to the specific cultivation method under thermophilic conditions. The susceptibility of Campylobacter isolates were determined for 13 antimicrobial agents. All enriched samples and cultured isolates were targeted for polymerase chain reaction (PCR) detection of 16S rRNA and multiplex PCR for determining their species. Among 302 fecal samples, 65 (21.5%) and 205 (67.8%) samples were positive for the presence of Campylobacter species with the cultivation and PCR techniques, respectively. All 65 distinct isolates were susceptible to neomycin and colistin and the isolates showed high resistance to cephalotin (83.0%) and ciprofloxacin (67.7%). After the multiplex PCR, 78.5% of total positive samples showed the simultaneous presence of Campylobacter jejuni and Campylobacter coli. In conclusion, the results emphasized that non-poultry farms are important as a possible source of Campylobacter infections.
    Keywords: Antibiotic resistance, Cattle, PCR, Sheep, Thermophilic Campylobacter spp
  • Masood Reza Seyfi Abad Shapouri, Maryam Ekhtelat*, Masood Ghorbanpoor Najaf Abadi, Mohsen Lotfi, Mohammad Rashno Pages 247-253
    Bovine Viral Diarrhea virus (BVDV) is an important viral pathogen of cattle causing several clinical syndromes. There are usually no pathognomonic clinical signs of BVDV infection. Diagnostic investigations therefore rely on serological detection and virus isolation. Nonstructural protein 3 (NS3) as immunogenic protein of BVDV is genetically and antigenically conserved among different isolates. This protein is therefore a candidate antigen for developing ELISA for serological studies. The aim of this study was to produce monoclonal antibody (MAb) against recombinant NS3 protein. For this purpose, the recombinant MBP-NS3 protein was expressed into expression vector pMalc2x in E. coli and purified using amylose resin chromatography column and the purified protein used as antigen in MAb production. After immunizing Balb/c mice with the recombinant antigen, the mouse showing highest titer of anti-NS3 antibody by indirect ELISA was selected for fusion. Spleen cells of the immunized mouse were fused with SP2/0 myeloma cells using polyethylene glycol. The cells in fusion mix were re-suspended in HAT medium and distributed in 96-well plates. Then culture supernatants of primary clones were screened by indirect ELISA. The positive clones after three times cloning, were selected and the reactivity of the monoclonal antibodies with recombinant and natural antigens was established by Western blotting. Based on these results, it appears that the specific monoclonal antibodies produced against NS3 recombinant antigen may be suitable for developing BVDV laboratory diagnostic assays.
    Keywords: Bovine viral diarrhea, Monoclonal antibody, NS3, Recombinant antigen
  • Raul Eduardo Pi, Ntildea., Aguilar*, Janet L., Oacute, Pez, Saucedo, Lilia Ivone Ruiz, Galaz, JosÉ De Jes, Uacute, S. Barroso, Padilla, Mayra Celina Gallegos, Rivas, Claudia Gonz, Aacute, Lez, Ortega, Antonio Martin GutiÉrrez, GutiÉrrez Pages 255-259
    Great apes are mammals close to humans in their genetic, behavioral, social and evolutionary characteristics and new genomic information is revolutionizing our understanding of evolution in primates. However, all these species are endangered. While there are many global programs to protect these species, the International Union for Conservation of Nature (IUCN) projects that in a near future the wild populations will decrease significantly. Nowadays, the relevance of captive populations of great apes is becoming critical for research and understanding of pathophysiology of diseases. In this report, the evaluation of infertility in a group of captive chimpanzees maintained at Leon’s Zoological Park using a human infertility protocol is described. Our results suggested that infertility in this group was due to low hormonal levels and sperm alterations in the male characterized by hormonal assessment and a sperm sample obtained by electroejaculation and cryopreserved using human protocols. In the females, it was demonstrated that it is possible to follow the follicular cycle using non-invasive methods based on morphological changes in genitalia, detection of blood in urine and measurement of hormones in saliva samples; concluding that fertility in females was normal. Also, we demonstrate that human artificial insemination procedures may be applied. Our human approach was successful in finding the infertility cause in this group of captive chimpanzees. In countries with limited resources, collaboration of zoos with human infertility clinics can be beneficial for research and management of reproductive aspects of great apes.
    Keywords: Chimpanzee, Cryptozoospermia, Cytogenetic analysis, Electroejaculation, Ultrasound
  • Fatemeh Neshat Halati, Alireza Vajhi *, Mohammad Molazem, Mohammad Mehdi Dehghan, Fereshteh Ansari Pages 261-266
    Dogs presented to the Small Animal Hospital of Veterinary Medicine, University of Tehran were included in the present study if spinal or intervertebral disc involvement was suspected. Clinical signs were recorded as well as general information of the patient such as age, breed and sex. Sixty dogs were examined radiographically and two standard orthogonal lateral and ventrodorsal projections were taken from the suspected region. Then magnetic resonance imaging (MRI) was performed for all patients. Agreement between MRI and radiographic findings, comparison of sex and breed with diagnostic imaging grades, comparison between diagnostic imaging grades and mean age, recovery rate after surgery or medical treatment, effects of diagnostic imaging severity grades on surgical or medical referrals were evaluated statistically. There were no significant association between age, sex and breed and frequency of the intervertebral disk disease. Intervertebral disc involvements between L2-L3 and T13-L1 were estimated as the most frequent sites of involvements. Sensitivity and specificity of radiography were evaluated 90.0% and 46.0%, respectively, by considering the MRI as a gold standard modality. There was a significant association between severity of disease in the MRI with referral to surgery and medical treatment. The recovery rate after surgery was significantly higher than medical treatment. These results can be used as a foundation for other studies with more focuses on details of injury and larger group of patients.
    Keywords: Dog, Intervertebral disk disease, Magnetic resonance imaging, Treatment
  • Adonis Pino*, Arlenis PÉrez, Aine Seavers, Guillermo Hermo Pages 267-270
    Caudal duplication (dipygus) is an uncommon pathologic of conjoined twinning. The conjoined malformation is classified according to the nature and site of the union. We report the presence of this malformation in a female crossbreed puppy. The puppy was delivered by caesarean section following a prolonged period of dystocia. External findings showed a single head (monocephalus) and a normal cranium with no fissure in the medial line detected. The thorax displayed a caudal duplication arising from the lumbosacral region (rachipagus). The puppy had three upper limbs, a right and left, and a third limb in the dorsal region where the bifurcation began. The subsequent caudal duplication appeared symmetrical. Necropsy revealed internal abnormalities consisting of a complete duplication of the urogenital system and a duplication of the large intestines arising from a bifurcation of the caudal ileum. Considering the morphophysiological description the malformation described would be classified as the first case in the dog of a monocephalus rachipagus tribrachius tetrapus.
    Keywords: Dipygus, Monocephalus, Puppy, Twinning
  • Soroush Sabiza, Ahmad Khajeh, Hadi Naddaf* Pages 271-274
    Tendon rupture in dogs is generally the result of a direct trauma. This report described the use of adjacent muscle autogenic fascial graft for reconstruction of distal rupture of long digital extensor tendon in a dog. A two-year-old male mix breed dog, was presented with a non-weight bearing lameness of the right hind limb and a deep rupture of lateral side of right tarsus. History taking revealed that this rupture appeared without any apparent cause, when walking around the farm, three days before. Radiography was done and no fracture was observed. Hyperextension of right tarsal joint compared to left limb was observed. Under general anesthesia, after dissections of the ruptured area, complete rupture of long digital extensor tendonwas revealed. Then, we attempted to locate the edge of the tendon, however, the tendon length was shortened approximately 1 cm. Hence, a strip of 1 cm length from fascia of cranial tibial muscle was harvested to fill the defect. The graft was sutured to the two ends of tendon using locking loop pattern. Subcutaneous layers and the skin were sutured routinely. Ehmer sling bandage was applied to prevent weight bearing on the surgical region. Re-examination and phone contact with the owner eight weeks and six months postoperatively revealed a poor lameness and excellent function of the dog, respectively. It could be concluded that the fascia of adjacent muscles can be used as an autogenic graft for reconstruction of some tendon ruptures.
    Keywords: Dog, Fascia, Graft, Rupture, Tendon