مجله تحقیقات دامپزشکی ایران (دانشگاه شیراز)
سال:1399 | دوره: | شماره: |تعداد مقالات:12

The process of fibrin clot formation is a series of complex and well-regulated reactions involving blood vessels, platelets, procoagulant plasma proteins, natural inhibitors, and fibrinolytic enzymes. Vasculitis can be caused by a variety of different agents as bacteria, viruses, protozoal, rickettsial organisms, toxic, drugs, medications, and neoplasms. The most common cause of vasculitis is the purpura hemorrhagica, which is associated with exposure to Streptococcus equi ssp. equi or less commonly, equine influenza. Deficiencies or defects of the hemostatic components may result in bleeding and/or thrombosis. Inherited alterations of primary hemostasis (von Willebrand disease: vWD and Glanzmann’ s thrombasthenia: GT) and of secondary hemostasis (hemophilia A and prekallikrein: PK deficiency) are scarcely reported in equine clinic. On the contrary, acquired alterations of primary and secondary hemostasis are commonly found. They include thrombocytopenia, platelet dysfunction due to the administration of some drugs and targeted antiplatelet agents, decreased factor synthesis (liver disease or deficiency of vitamin K), release of inactive factors, inhibition of factor activity, or excessive consumption and depletion of factors (platelets, coagulation factors, and anticoagulants factors as antithrombin (AT) and protein C). Disseminated intravascular coagulation (DIC) is the most common and complex hemostatic disorder in horses and appears to be associated with sepsis, inflammatory and ischemic gastrointestinal tract disorders and other systemic severe diseases. These alterations are commonly found in patients in intensive care units.

Background: Despite the popularity of ice cream, its high content of sugar and fat is worrisome. Substituting sugar with natural and useful resources is one possible solution. Dates (Phoenix dactylifera) are rich in carbohydrates and are a good source of energy; they also have a large amount of phenolic compounds and anthocyanin. Aims: The aim of this study is to evaluate the effect of ice cream sugar replacement with date products. Methods: Accordingly, four ice cream samples with different sweeteners including 1) sucrose (as a control), 2) date pulp + sucrose, 3) date liquid sugar, and 4) sucrose + date liquid sugar, were produced. Total phenolics, anthocyanins, and antioxidant activities of dates and their effects on physicochemical and sensory properties of ice cream were investigated. Results: A large amount of phenolic compounds and anthocyanin was found in the ice cream contained date pulp and liquid sugar, which enhanced the reducing power and antioxidant effect of it. The overrun and density of an ice cream did not change with sugar replacement. Total substitution of sucrose by date liquid sugar, increased the titratable acidity, viscosity, and antioxidant properties of ice cream, but it was found to have some negative effects on the organoleptically score of color. However, the partial replacement of granular sugar with date pulp/liquid sugar, improved the antioxidant value of this dairy dessert without any change in its color. Conclusion: It seems that the addition of date pulp to an ice cream, not only maintain the quality and sensory properties of an ice cream, but also provides an antioxidant property.

Background: Pneumonia due to Mycoplasma infections can cause serious health problems and economic losses in small ruminants industry. Aims: The aim of this study was isolation and identification of Mycoplasmas in sheep naturally infected with pneumonia in Northeastern Iran. Methods: This study used histopathology, culture and polymerase chain reaction (PCR) to examine samples from 50 lungs of sheep naturally infected with Mycoplasmas. Results: Grossly, irregular consolidation with lobular or lobar to diffuse pattern in the cranioventral to caudal lobes of affected lungs were observed. Histopathologically, bronchointerstitial pneumonia in 38 (76%), and purulent to fibrinopurulent bronchopneumonia in 12 (24%) affected sheep were diagnosed. DNA was extracted from lung tissue samples and replicated using genus and species specific primers for Mycoplasma. Mycoplasma growth was observed in 3 (6%) of a total of 50 lung samples. Genus-specific Mycoplasma DNA was identified by PCR in 12 (24%) of samples. Two (4%) and 7 (14%) samples of these 12 cases were positive for reaction with species-specific primers of Mycoplasma ovipneumoniae and Mycoplasma arginini, respectively. Conclusion: Our results showed that M. ovipneumoniae and M. arginini were the two agents that can be involved in inducing lung consolidation and pneumonia in sheep and PCR was more successful than the culture in detecting Mycoplasmas.

Background: Salmonella can cause serious human gastroenteritis and is frequently isolated from various food samples. The cell culturing, immunoassay, and polymerase chain reactions (PCR) are the current methods to detect such pathogenic agents. However, these methods are time-consuming and labor-intensive, and thus unavailable for rapid-monitoring of Salmonella. Aims: This study aimed to develop an immunocapture-loop-mediated isothermal amplification (IC-LAMP) for rapid and sensitive detection of Salmonella. Methods: Salmonella was used as antigen to produce monoclonal antibody (mAb) and mAbs were prepared via subcloning three times. The mAb 1B12 with high affinity was coated on the surface of the immuno-magnetic beads (IMBs) to capture Salmonella. The enriched products (IMBs-Salmonella) were used for LAMP using the special primers targeted the conserved invA gene of Salmonella. Results: The IC-LAMP was developed based on mAb 1B12 and LAMP. Targeting the conserved invA gene of Salmonella, the detection time was shortened to 50 min from three days. If the reaction contains Salmonella, the green fluorescence and the trapezoidal strip can be clearly observed. Importantly, the method combines the specificity of antibody and LAMP with a detection limit of 5 CFU/ml in artificially contaminated water and milk. The specificity of this method was demonstrated by testing other similar bacteria. The results indicate that the IC-LAMP reacts only with Salmonella and does not crossreact with other similar bacteria. Conclusion: The IC-LAMP assay developed here is a rapid, sensitive, one-step-visual method to screen for the presence of Salmonella in food samples. This method is faster than traditional PCR, LAMP, and other methods, and can be used as a primary screening method for the detection.

Background: Lactococcus garvieae causes lactococcosis in rainbow trout in many parts of the world. Aims: This study was conducted for the existence of the virulent factors and differentiation of the two serotypes in L. garvieae. Methods: Twenty-two strains of L. garvieae isolated from diseased rainbow trout from farms in different regions and mugger crocodile of Iran, were investigated. In order to rapidly detect the presence of the hly1, hly2, hly3, NADH oxidase, sod, pgm, adhPsaA, eno, LPxTG-3, adhCI, and adhCII virulence genes, two multiplex polymerase chain reaction (PCR) assays were developed. Also, simplex PCR method was used to identify the bacterial serotypes, CGC, LPxTG-2, Adhesion, and adhPav virulence genes using the specific primer. Results: All varieties of L. garvieae contained the hly1, hly2, hly3, NADH oxidase, pgm, adhPav, LPxTG-3, sod, eno, adhPsaA, adhCI, and CGC virulence genes. Also, adhCII gene was present in all strains except one of the isolates originated from mugger crocodile. In addition, LPxTG-2 gene was only present in one of the isolates belonging to mugger crocodile. Adhesion gene was not present in all the strains. Interestingly, all the 22 strains originated from both hosts were identified as belonging to the serotype I. Based on the phylogenetic sequences of the capsule gene cluster, group all fish isolates into a cluster together with one isolate obtained from mugger crocodile. Conclusion: Further studies are recommended to investigate the role of virulence genes in L. garvieae and evaluate their pathogenicity to rainbow trout.

Background: Infectious bronchitis virus (IBV) is the etiological agent of an acute and highly contagious disease. Infectious bronchitis (IB) affects chicken of all ages and poses major economic loses to the poultry industry worldwide. The continuous evolution of the spike protein (S1) of IBV is responsible for the prevalence of many serotypes/genotypes around the world. Multiple lineages of IBV strains have been detected in chicken flocks in India since 2003. Aims: To detect IBV genotypes prevalent in India. Methods: Organ samples from 20 IBV-positive flocks with variable clinical signs were used for the amplification of the S1 gene of IBV by reverse transcriptase-polymerase chain reaction (RT-PCR). Results: Positive PCR amplicons were sequenced. Sequence analysis showed that 14 field isolates belonged to the GI-1 genetic lineage (Mass 41 serotype), two field isolates belonged to the GI-13 (UK 4/91 variant IBV strain), one field isolate grouped with GIII, GV, and GVI genetic lineage and three belonged to a variant genotype unique to India (GI-24). Phylogenetic analysis also showed a similar type of grouping within the field isolates. Among the fourteen GI-1 isolates, 12 were isolated between 2003 and 2006 and only two were isolated between 2009 and 2011. The two field isolates belonging to GI-13 were isolated in 2007, another one belonging to GIII, GV, and GVI was isolated in 2010 and three field isolates were not close to any reference IBV sequences isolated in 2006 (IND-TN-168-06), 2010 (IND-TN-280-10) and 2011 (INDTN-290-11). Conclusion: A unique variant of IBV is emerging in India (GI-24). Our findings will have important implications for future vaccine intervention.

Background: Canine hip dysplasia (CHD) is an orthopedic disorder characterized by abnormal laxity of the hip joint. It is considered multifactorial and polygenic and affects predominantly medium and large sized dog breeds. Aims: The aim of this study was to identify CHD associated polymorphisms in chromosomal regions on CFA19, CFA24, CFA26, and CFA34. Methods: Blood samples from 60 dogs of different breeds were collected and genotyped, including 46 cases and 14 controls. After sequencing and single nucleotide polymorphism (SNP) determination of the target regions, an individual SNP analysis with a  2 statistic was performed based on the comparison of allele frequencies in cases and controls. Results: A significant association was observed between CHD and a T/C SNP on CFA19, which harbors genes involved in bone metabolism. No other significant association was found in the study and previously identified SNPs cannot be validated as related to CHD. Conclusion: Further research is warranted to identify CHD-associated polymorphisms in order to develop a genotype-based diagnosis and selection approach.

Background: Implantation is a crucial period determining the success of a full pregnancy. Endocrine disruptors such as phytoestrogens (PEs) were thought to adversely influence embryonic implantations. However, the mechanism by which they upset implantation was not fully elucidated. Aims: The effect of administering soy isoflavones on the implantation of Wistar rats was studied through the detection of progesterone receptors (PR), vascular endothelial growth factor (VEGF), and estradiol receptor alpha (ER-α ) protein expression at gestation day 6 (GD6). Methods: Eighteen cyclic female Wistar rats were distributed into two groups, group A: control (n=9) were fed with a casein based diet, and group B (n=9) were fed with a casein diet and gavaged 50 mg/kg/day soy isoflavones’ extract 40% starting from gestation day zero (GD0) to GD6. Feed intake, body weight (BW), body gain, and uterine weights were recorded. At the end of GD6 the number of corpora lutae (CLs) and implantation rates were recorded. Histopathology and immunohistochemistry (IHC) for PR, VEGF, and ER-α protein expression in implanted uteri were performed. Results: Soy isoflavones significantly reduced feed intake, weight gain, uterine weights CL numbers, and implantation rates of the treated pregnant dams. The endometrium of the soy treated dams showed less proliferation than that of the control. Immunostaining percentage of PR and VEGF proteins significantly reduced in soy treated dams compared to the control. However, the mean expression percentage of ER-α exhibited significant elevation in the soy treated dams in comparison to the control group. Conclusion: Implantation losses caused by soy isoflavones seemed to be due to the down regulation of PR that failed to down regulate ER-α action and decreased VEGF production.

Background: Paratuberculosis and tuberculosis (TB) caused by Mycobacterium avium paratuberculosis (MAP) and Mycobacterium tuberculosis complex (MTC), respectively are economically important, chronic debilitating diseases affecting the dairy herds and are also potential zoonotic threats. Aims: Differential diagnosis of paratuberculosis and TB in blood samples of cattle and buffaloes. Methods: In this study, an in-house developed multiplex polymerase chain reaction (PCR) targeting MAP, Mycobacterium bovis and Mycobacterium smegmatis was used in blood samples (buffy coat) parallel with IS900 PCR and esxB PCR for diagnosis of paratuberculosis and TB, respectively; in a total of 202 cattle and buffaloes. Results: Out of 202 animals, 12 (5. 9%) and 17 (8. 4%) animals were positive for MAP by multiplex PCR and IS900 PCR, respectively; from which only 8 (4%) animals were positive by both tests; whereas 4 and 9 animals were exclusively positive by multiplex PCR and IS900 PCR, respectively. None of the animals were found to be positive for M. bovis and M. smegmatis by the multiplex PCR. However, the esxB PCR detected 13 (6. 4%) animals positive for TB. In fact, 3 (1. 5%) animals were found to be co-infected by both paratuberculosis and TB. Conclusion: The in-house multiplex PCR detected MAP in buffy coat and there was a fair degree of agreement between the multiplex PCR and IS900 PCR in detection of MAP DNA though the latter detected more number of animals to be positive for MAP. Besides, esxB PCR showed a high diagnostic potential and can be used for diagnosis of TB from blood.

Background: Circoviruses are small, non-enveloped, single stranded DNA viruses. There is scarce information about these agents in non-psittacine birds. Aims: It is attempted to detect and characterize circoviruses in non-psittacine birds. Methods: Forty-five samples were collected from different non-psittacine species belonging to seven avian orders. A nested polymerase chain reaction (nested-PCR) for the detection of rep gene of circoviruses was applied. Results: Two different types of circoviruses were detected in two pigeon samples (2/11, 18. 2%). One of the detected circoviruses was placed in clade A next to a polish strain based on phylogenetic analysis. Interestingly, the other detected circovirus was closely related to canary circoviruses (CaCVs). Conclusion: In addition to the molecular diagnosis of a pigeon circovirus (PiCV), this is the first report of the detection of CaCv in a pigeon. The possible hypotheses of such circumstance are discussed.

Background: Aortic thrombosis (ATh) is an uncommon problem in dogs. Although the pathogenesis of hypertrophic osteopathy (HO) is unknown, it is thought this can be due to blood flow disorder. In this case, removal of aortic thromboembolism (ATE) resulted in periosteal proliferation. Case description: A 4. 8-kg, 3-year-old, intact female Maltese was referred with a bite wound showing pneumomediastinum, pneumothorax, and subcutaneous emphysema. After adequate treatment, the dog had recovered well and was discharged. Findings/treatment and outcome: Acute non-painful paraparesis suddenly developed 5 days after discharge. An abdominal ultrasound showed ATE at the level of 1 cm proximal to the external iliac arterial bifurcation. Based on clinical sign, physical exam, neurologic exam, and ultrasonography, ATh was diagnosed. Arterial thrombectomy was performed to remove the thrombus. Twenty-four days after surgery, the dog had pain and soft tissue swelling of both stifles due to HO. After rehabilitation, the dog finally started to ambulate, and the dorsal pedal arteries pulse was normal. Hypertrophic osteopathy also resolved completely. Conclusion: This report suggests the relationship between HO and ATh, for the first time in veterinary medicine.

Background: Splenic infarction (SI) is a rare clinical entity seldom encountered in veterinary medicine. Its most frequent causes include thromboembolic status, splenomegaly, and cardiac disease. Although thrombotic elements from the circulation provide the most common context for thromboembolic SIs, immune-mediated hemolytic anemia (IMHA) has not been reported as an underlying disease in canine SI. Case description: A 2-year-old, female spayed Dachshund, was referred with vomiting, hematochezia, and brown colored urine over the preceding 4 days. Physical examination revealed abnormalities including generalized weakness, jaundice, and splenomegaly; blood work showed pancytopenia and hyperbilirubinemia. Erythrocyte agglutination, polychromasia, and spherocytes on a peripheral blood smear were observed and IMHA concurrent with thrombocytopenia was diagnosed. Findings/treatment and outcome: Although erythrocyte agglutination and leukopenia disappeared after treatment, anemia and thrombocytopenia were unresponsive to oral immunosuppressive drugs and repeated transfusions. Further abdominal ultrasound identified an occlusive splenic vein thrombus. Splenic histopathology found marked multifocal to coalescing necrosis, and hemorrhage consistent with multiple SI. Symptoms resolved following splenectomy combined with 1 month of immunosuppressive medication, and the dog was healthy on follow-up evaluation after 2 years. Conclusion: Immune-mediated hemolytic anemia is an incompletely characterized cause of SI. This report establishes a potential and novel causal role for IMHA in canine SI. We believe it to be the first case report of SI in a dog with refractory IMHA and thrombocytopenia, successfully managed by splenectomy combined with short-term immunosuppressive therapy.