Volume 65 (2015): Issue 1 (March 2015)

Spontaneous hypercortisolism or Cushing's syndrome is a common endocrinopathy in dogs. Pituitary-dependent and adrenal-dependent hypercortisolism each require specific treatment and diagnostic imaging is very helpful in choosing the treatment that is appropriate. The aims and expectations of the treatment need to be established beforehand and discussed with the owner to avoid unexpected disappointments. The clinical signs of pituitary-dependent hypercortisolism caused by a pituitary microadenoma can be managed with the adrenocorticostatic drug trilostane, but the drug will not affect the pituitary tumor. Hypophysectomy is therefore preferred in those dogs that have an enlarged pituitary but are in good clinical condition and have a long life-expectancy. Inoperable pituitary tumors can be treated by radiotherapy. The best treatment in dogs with cortisol-secreting adrenocortical tumors is adrenalectomy. If surgery is not possible, because of vascular invasion or metastatic spread, mitotane is recommended. Treatment with trilostane can be considered but is only palliative: it does not affect the adrenocortical tumor.

A simple and rapid real-time loop-mediated isothermal amplification (LAMP) assay designed to detect Lawsonia (L.) intracellularis, an important bacteria causing proliferative enteropathy in pigs. A set of four primers targeting the ubiquinone/menaquinone biosynthesis methylase (ubiE) gene was designed for the LAMP reaction. Additionally, serial 10-fold dilutions of cultured L. intracellularis and spiked feces were also used for the optimization of real-time LAMP. The lower limit of the linear range of the assay in L. intracellularis was 1.0 × 10⁰L. intracellularis. Real-time LAMP was 10 and 100 times more sensitive than real-time PCR and conventional PCR detection methods, respectively. Based on testing of 213 porcine fecal samples using real-time LAMP, realtime PCR and PCR, the agreement quotients of real-time LAMP with conventional PCR and with real-time PCR were 0.77 and 0.95, respectively. This study demonstrated that real-time LAMP was a powerful tool for the rapid and sensitive detection of L. intracellularis in porcine fecal samples.

The study was undertaken considering: i) that relative proportion of distinct subsets of splenic dendritic cells (DCs) is strain-specific and predictive for the susceptibility to autoimmune diseases; ii) age-related changes in endocytic, allostimulatory and polarizing capacity of splenic OX62+ DCs from Albino Oxford rats (relatively resistant to Th1/Th17-mediated diseases) and iii) strain specificities in age-related changes of mouse DCs. To ascertain whether there are strain specificities in age-related rat DC changes, we examined the influence of aging on OX62+ DCs from Dark Agouti (DA) rats prone to Th1/Th17-mediated autoimmune diseases. The study provided additional evidence that the predominance of CD4-cells within OX62+ DCs from young adult rats correlates with their susceptibility to Th1/Th17-mediated diseases. Consistently, lipopolysaccharide (LPS)-matured DCs from 3-month-old (young) rats exhibited Th1 driving force when co-cultured with allogeneic CD4+ T cells. This most likely reflected enhanced TNF-α and iNOS expression. Comparing with young rats, OX62+ DCs from 26-month-old (aged) rats showed: i) diminished endocytic capacity; ii) impaired ability to mature in vitro upon LPS stimulation (as indicated by lower MHC II, CD86 and CD40 surface expression), which is consistent with the increase in their IL-10 production, and iii) diminished allostimulatory capacity and loss of Th1-driving capacity in the mixed lymphocyte reaction. The latter, probably, reflected greater IL-10 production by LPS-stimulated DC from aged rats, as well as lower CD40 density on their surface. Overall, our findings suggest that aging might affect DA rat capability to mount an efficient Th1 immune response, and consequently susceptibility to Th1/Th17-mediated pathology.

The purpose of the present study was to evaluate serum canine Cystatin-C (Cys-C) concentration for the prediction of renal function (RF) in dogs naturally infected with Babesia canis vogeli and Ehrlichia canis. A total of 46 dogs were enrolled into three groups. Group B included 16 dogs naturally infected with B. c. vogeli, group E 10 dogs naturally infected with E. canis [dogs diagnosed as canine monocytic ehrlichiosis (CME)], and group H involved 20 healthy controls (negative for B. c. vogeli and E. canis). Solely dogs presenting uncomplicated babesiosis caused by B. c. vogeli were enrolled. Serum urea and serum creatinine (Cre) levels with serum Urea:Cre ratios were analyzed. Canine serum Cys-C was determined with a species-specific commercially available and validated ELISA assay as a reference. Mean serum Cys-C levels were 5.28 mg/L, 3.02 mg/L, and 2.30 mg/L for groups B, E and H, respectively. RF in uncomplicated CB caused by B. c. vogeli seems to be affected based on serum Cys-C concentrations. To the author's knowledge, this is the first study reporting that mean serum canine Cys-C levels in dogs with CME demonstrated no elevation. Serum canine Cys-C as a novel RF marker could be used for early detection of renal injury in both babesiosis and CME.

The aim of this study was to determine whether synovial fluid (SF) of dogs contains cells that have characteristics of MSCs and to describe their differentiation potential. SF adherent cells from 5 young German shepherd dogs (average 3.8 ± 0.9 years) were expanded (37°C, 5% CO₂, humidified atmosphere) three weeks before their phenotype was characterized by flow-cytometry for the presence of CD90 and CD34. Population doubling time (PDT), number of CFU-F and adipogenic, osteogenic and chondrogenic potentials have been determined in vitro. In early passages PTD was 31 ± 10 hours and expansion fold after 3 sub cultivations (9 days) theoretically could be 372 ± 134. At P1, 0.55 ± 0.05% of SF cells had the ability to form CFU-F. Sixty-six percent of cells expressed CD90 and none of the cells expressed markers of hematopoietic cells. Oil Red O staining has shown accumulation of fat droplets in cells grown in adipogenic medium, while deposits of calcium in the osteogenic medium were evidenced with Alizarin red staining. SF cultured in hondrogenic and control medium in three-dimensional conditions formed a cartilage-like tissue. Alcian blue staining of pellets’ slides have shown a significant amount of glycosaminoglycans (GAGs) and immunohistochemistry analysis documented collagen type II expression. The amount of GAGs in pellets grown in both conditions showed no difference. SF cells in vitro exhibited osteogenic, adipogenic and chondrogenic differentiation potentials, suggesting the presence of different mesenchymal progenitors. These results also demonstrated that SF cells have a spontaneous chondrogenic potential that should be further explored for possible tissue engineering protocols.

A retrospective study on 235 natural cases of Porcine Respiratory Disease Complex in order to determine the etiological agents, their prevalence and interrelationships was performed in Serbia. Lung tissue samples were analyzed by Polymerase Chain Reaction for the presence of Porcine circovirus type 2, Porcine reproductive and respiratory virus, Swine influenza virus, Mycoplasma hyopneumoniae, Pasteurella multocida, Actinobacillus pleuropneumoniae, Haemophilus parasuis, Streptococcus suis and Arcanobacterium pyogenes. A total of 49 different combinations of viral and bacterial pathogens were found. Five different viral and viral/Mhp co-infections were detected. Monobacterial infections were found in 150 cases and polybacterial infection was detected in 85 samples. PCV2 was the main virus detected, and Pm was the most aggressive secondary pathogen detected in PRDC. The reason for PRDC being so prevalent among Serbian pigs is most likely due to the large number of risk factors in the conventional farrow-to-finish system, compared to multi-site production systems. Therefore, measures aimed at a better control of respiratory viruses, particularly Porcine circovirus type 2 and Porcine reproductive and respiratory virus, as well as Mycoplasma hyopneumoniae infections, and adoption of rational decisions on respiratory bacterial pathogens specific therapeutic and preventive strategies at herd level, simultaneously with significant improvements on farm management should reduce the occurrence of PRDC.

The aim of the study is to increase the knowledge on the genetic structure status of the autochthonous Slavonian Syrmian Podolian cattle breed currently listed by the FAO as “critically endangered”. Blood samples obtained from a total of 20 randomly selected individua were included in genotyping upon microsatellite panel recommended by ISAG and FAO. The procedure enabled the identification of as many as 214 (96.34%) genotypes and, at the same time, 129 allelic variations were found with = 11.73 alleles per locus. The genotypes showed differences between the microsatellite loci and also significant differences (p<0.05) in the frequency of both homozygosity and heterozygosity. The analyses of the allelic distribution within microsatellite loci showed marked variability. The distribution of genetic relationship was expressed by Wright's F coefficients. A moderate level of genetic diversity was observed at the tested loci. The findings demonstrate the degree of breed relationship and breed purity but also indicate that gene pool of the tested population has been threatened. Therefore, urgent measures must be taken to support both the genetic consolidation and preservation of the remaining population of Slavonian Syrmian Podolian cattle in Croatia.

In this study the seroprevalence of anti-Borrelia burgdorferi IgG antibodies in populations of naturally exposed dogs in the Belgrade area was determined. Serum samples of 215 dogs were tested with in-house made indirect immunofluorescent-antibody assay (IFA), enzyme-linked immunosorbent assay (ELISA), and Western blot assay (WB). Sera were from 34 pet dogs suspected of having Lyme borreliosis, 41 hunting dogs, 125 shelter dogs, and 15 clinically healthy pet dogs. Results of WB were used as a standard against which the performances of ELISA and IFA were evaluated. ELISA was significantly more sensitive than IFA (84.8 vs 67.9%), whereas the specificities of these tests were just slightly different (94.4% vs 92.6 %). Borrelia burgdorferi seroprevalence of 24.7% among tested dogs was recorded by WB test. The highest seroprevalence of 31.2% was found in the group of shelter dogs, while hunting and pet dogs showed lower seroprevalence of 19.5% and 17.6%, but the difference was statistically insignificant. These results are the first report on the seroprevalence of IgG antibodies to B. burgdorferi in dogs in the Belgrade area and could be of importance in the assessment of the risk of infection with B. burgdorferi in human population.

The purpose of this study was to evaluate in calves the effects of respiratory diseases of various severities on blood gases, acid-base balance and plasma lactate concentration. We included in the study 128 calves with clinical signs of various severities and outcomes of respiratory disorders. The age of the calves ranged from 1 to 12 months. According to the clinical findings, course of the disease and time of blood collection, the sick calves were divided into three groups: RD 1 (n=34) – calves with moderate clinical signs; RD 2 (n=73) – calves with severe respiratory symptoms and RD 3 (n=21) – calves with severe respiratory symptoms and died within 48 hours after blood sampling. Into the study we also included one group of 28 clinically healthy calves. In the arterial blood we evaluated: blood pH, partial pressure of carbon dioxide – pCO₂, partial pressure of oxygen – pO₂, concentration of actual bicarbonate – HCO₃⁻, base excess – BE, and saturation of hemoglobin by oxygen – O₂-sat. The concentration of lactate was determined in the blood plasma. Significant effects of health status and severity of respiratory diseases were recorded in all the evaluated variables. Marked hypoxemia and hypercapnia were found in calves with severe respiratory disorders and in animals that died within 48 hours after blood collection. In most calves, the acid-base changes were characterized as compensated respiratory acidosis. The highest mean lactate values were recorded in calves that died within 48 hours after blood sampling. The presented results suggest the usefulness of the analysis of blood gases and acid-base status in the evaluation and prognosis of various degrees and severity of respiratory diseases in calves, and in monitoring the patient response to treatment. Although plasma lactate analysis is less consistent with the severity of respiratory diseases, it might be helpful in the indication of an untreatable stage of the disease and upcoming death.

The aim of the study was to establish morphometric, biochemical and hematological values for the endangered Balkan donkey breed (Serbia) and to explore the possible age dependence of the parameters tested. Inter-breed similarity of morphometric parameters was assessed by comparing the data obtained for the Balkan donkey with morphometric measurements of several previously characterized domestic donkey breeds. The study population included 74 donkeys, divided in two age groups (group A ≤ 3 years; group B > 3 years). In total, 18 morphometric, 13 hematological and 14 biochemical parameters were assessed. Significant morphometric differences (p<0.05) in body length, head length, chest circumference and body weight were found between the two age groups. Significant differences in morphological parameters were revealed among the Balkan donkey and other donkey breeds (Catalonian, Croatian and Albanian), but results of cluster analysis demonstrated the smallest distance between the Balkan donkey and Albanian donkeys. The results of morphometric analyses showed consistency of the obtained values within the breed, and diversity as compared to other donkey breeds, and, thus, could be taken as referent for the Balkan donkey. Hematological and biochemical profiles obtained for the Balkan donkey were consistent with previous reports and within the recommended reference ranges. White blood cell, mid cell and granulocyte counts, showed significantly higher (p<0.05) values in donkeys under 3 years of age, while the only biochemical parameter affected by age was alkaline phosphatase. The information gained through characterization of the Balkan donkey breed provides a basis for conservation and development of the breed standard.

A ten year old mongrel bitch was diagnosed with a primary vulvar transmissible venereal tumor (TVT) causing subcutaneous, mammary and splenic metastasis and it was successfully treated with vincristine. Four months later the animal presented neurological disturbances, brain metastases were suspected and the animal was euthanized. A TVT metastatic mass was found in the brain and confirmed with immunohistochemical results, being positive for vimentin and lysozyme and negative for S-100, CD3 and cytokeratin. TVT metastases in the brain are a rare event and cannot be treated with usual chemotherapy as vincristine does not cross the blood-brain barrier, thus allowing the re-emergence of the tumor after a period of time.

A two-year old Jack Russell Terrier female with left-sided epistaxis was brought to the Clinic of the Bucharest Faculty of Veterinary Medicine. Endoscopy of the left nasal cavity revealed a cauliflower-like mass. Cytological and histopathological features were specific to canine transmissible venereal tumour (CTVT). Tumor tissue showed positive immunoreactivity to anti-vimentin monoclonal antibody (90%) and Ki67 (35%) and samples were negative for anti-cytokeratin monoclonal antibody. Chemotherapy with vincristine sulphate at a dose of 0.025 mg/kg i.v./week for 6 administrations was successful. This paper presents a primary form of nasal CTVT in a Jack Russell Terrier female.