Volume 66 (2016): Issue 4 (December 2016)

Although Mycobacterium avium subspecies are generally not considered food pathogens, the infections caused by these particular nontuberculous mycobacteria (NTM) can represent a serious threat to immunocompromised population. Additionally, infections with a member of Mycobacterium Avium Compex (MAC) can affect the efficiency of BCG vaccines used for the humans. In infected animals, M. avium may be present in different tissues without apparent clinical symptoms and macroscopic lesions. Veterinary meat inspection would then fail to recognize infected animals and such meat and meat products thereof could enter the human diet. The aim of this paper is also to analyze the current control policy in Europe according to infections of pigs with the members of MAC, and point out the risks for public health. By analyzing a large number of meat samples and other dietary nutrients, different groups of authors have provided evidence to support the hypothesis that M. avium is present in the everyday environment. Therefore, food as a source of infection with mycobacteria should not be ignored. The control of mycobacteria requires a better diagnostic approach, having in mind recent positive cases of M. avium subspecies hominissuis (MAH) in an increasing number of exported pigs from EU countries to Serbia. The introduction of reliable diagnostic methods for MAH could result in decreasing the occurrence of infection in pigs, as well as in humans, having in mind that WHO reported 10 million new cases of tuberculosis-mycobacteriosis in the human population in 2015 with 21% of these cases occurring in immunocompromised individuals and children.

Lumpy skin disease (LSD) is an important disease of cattle which is included in the OIE list of notifiable terrestrial animal diseases because of its great economic importance. The etiological agent is the Lumpy skin disease virus (LSDV).

In the control of LSD attenuated strains of LSDV and SPPV are successfully used as vaccine strains in infected areas. In the case of vaccination policy, due to the possibility of mild or systemic post-vaccination reactions in vaccinated animals, the application of diagnostic procedures that will rapidly and specifically differentiate LSDV field strains from LSD vaccine virus strains are extremely important. Rapidity in diagnostics and disposal of infected animals is one of the key factors in the prevention of spreading the disease.

In the presented study we have described the development and validation of two real-time TaqMan-PCR assays for a rapid, sensitive and specific detection of the virulent field LSDV strain currently circulating in the Balkan Peninsula. Specificity for the field strain and exclusivity for vaccine strains was tested on 171 samples from naturally infected and vaccinated animals.

The results of this study show that both developed real-time PCR assays are more sensitive than the conventional nested PCR in detecting field LSDV strains thus enabling rapid and high-throughput detection of animals infected with field strains of LSDV.

In conclusion, both KV-2 and FLI real-time PCR assays described in this study are simple, rapid, sensitive and suitable for routine use in a diagnostic laboratory and have the potential to replace conventional nested gel-based PCR assays as the standard procedure for the detection of field strains of LSDV in clinical samples.

The aim of this study was to re-evaluate archived samples of canine soft tissue sarcomas (STSs) morphologically consistent with peripheral nerve sheath tumors (PNSTs). In each case, an immunohistochemical panel was applied, including α-SMA, calponin, desmin, S-100, GFAP, NSE and Olig2, in order to assess whether the phenotype was consistent with the tumor histological appearance. Additionally, the expression of EGFR, a marker with potential therapeutic implications in malignant PNSTs, was evaluated. Twenty-one tumors were included. Fourteen cases (66.7%) were positive for one or more muscular markers and were reclassified as perivascular tumors (PWTs). A positive labeling for S-100 was observed in one tumor (4.8%), thus classifi ed as PNST. The other 6 tumors were generically classified as poorly differentiated STSs. No unique histopathological feature was observed within the three groups. NSE and Olig2 labeling was aspecific and not useful for diagnostic purposes. GFAP was negative in all cases. Six cases (28.6%) were positive for EGFR, including the PNST. Even after the application of a wide immunohistochemical panel, distinguishing between PNSTs and PWTs remains a challenge. Finally, a subgroup of cases cannot be classified based on light microscopy alone.

Staphylococcus aureus is known worldwide as a frequent cause of mastitis in dairy cattle. Due to the production of heath resistant enterotoxins, this pathogen is also a major cause of food poisoning among humans, with symptoms of often severe vomiting and diarrhea. The aim of our study was to determine the prevalence of enterotoxinproducing strains of S. aureus originating from samples of cows with subclinical and clinical mastitis in the Republic of Serbia. Furthermore, we analyzed the type of staphylococcal enterotoxin they produce and phylogenetic relatedness among the S. aureus isolates recovered from milk in this study. Production of staphylococcal enterotoxins A, B, C, D and E was determined by commercial immunoenzyme assay VIDAS® SET2, and presence of corresponding genes encoding enterotoxin synthesis in positive isolates confi rmed by Polymerase Chain Reaction. Enterotoxin production was determined in 5 out of 75 (6.67%) isolates of S. aureus and all of them produced staphylococcal enterotoxins C. After analyzing the nucleotide sequence of the gene encoding the synthesis of staphylococcal protein A, S. aureus isolates were assigned into 2 phylogenetic groups, including 7 clusters. All S. aureus isolates with the presence of sec gene formed one cluster even dough they originated from milk samples from different farms.

Carvacrol is a monoterpenic phenol and an active ingredient of the plant essential oils of the family Lamiaceae. We have investigated the analgesic effect of carvacrol, the possible dependence of the effect in relation to animal sex, and the impact of carvacrol on motor coordination in rats. Hyperalgesia was induced by formalin (1.5%), which was administered SC in the upper lip of rat. Hyperalgesia and effects of carvacrol and indomethacin were measured by using the orofacial formalin test. The influence on motor coordination in animals treated with carvacrol was investigated by using the rota-rod test. Carvacrol administered PO in pre-treatment (45 min. prior to formalin) at a single dose of 50, 75 and 100 mg /kg BW, in the male, 50 and 100 mg /kg BW, in female rats caused a dose-dependent antinociceptive effect. This effect of carvacrol was significantly higher (P<0.01, P<0.001) in male rats. Compared with indomethacin administered during pre-treatment (2 mg/kg, PO), carvacrol (100 mg/kg) exhibits significantly higher (P <0.05 and P <0.001) antinociceptive effect on formalininduced hyperalgesia in male rats. In the rota-rod test carvacrol did not disturb the motor coordination in male rats, nor the dose of carvacrol with clear antinociceptive properties exhibited depressive effect on the CNS of treated rats. Keeping in mind that the monoterpene carvacrol is of plant origin, with potentially less side effects and without residues, it is realistic to expect the possibility of its therapeutic use in the treatment of inflammatory pain in animals.

The aim of this study was to assess whether atipamezole can restrain telazol/xylazine induced expression of c-fos in the rat brain. Rats were injected with a mixture of 13.81 mg/kg telazol and 5.21 mg/kg xylazine, following 10 min later 0.522 mg/kg atipamezole. Thereon, the thalamencephal and cerebral cortex were removed one hour after the last injection. The level of Fos protein was measured in the brain tissue by Westernblot. The results revealed that atipamezole attenuates telazol/xylazine induction of c-fos expression in the thalamencephal and cerebral cortex. The results indicated that atipamezole is able to inhibit telazol/xylazine-induced c-fos expression in the rat brain, thus protecting it from nerve damage.

The purpose of the present study was to investigate and compare the effects of two ruthenium complexes with trifluoperazine on acethylcholinesterase enzyme activity and lactate dehydrogenase levels in vivo under physiological conditions in rats blood. Complexes 1 and 2 showed positive effects on acethylcholinesterase at all doses and did not disturb its normal activity. Total LDH activity was inhibited in the presence of both complexes, but Ru(II) complexes showed different effects on the activity of LDH isoenzymes. The activities of LDH₁ and LDH₂ isoenzymes were decreased in all applied doses of the complex 2, while the activity of LDH₂ reduced using complex 1 in the same doses. Results of the present study suggest the neuro- and cardio protective potential of oral administration of complexes 1 and 2, as non-toxic compounds under physiological conditions. These protective effects are the result of their potent antioxidant activity.

The presence of bovine parainfluenza virus type 3 (BPIV3) was examined in 119 nasal swabs collected from cattle with severe respiratory infection. All samples were conducted for virus isolation on the MDBK cell line. The cytopathic effect was observed after 48h to 72h in cells inoculated with eight samples (8/119; 6.7%). The confirmation of isolated strains of BPIV3 was done by the virus-neutralization test. In addition, all samples of bovine nasal swabs were also examined for the presence of BPIV3 virus using RT-PCR with primers specific for the part of HN gene. The presence of BPIV3 was detected in eight samples (8/119; 6.7%) that were also positive upon virus isolation. The molecular characterization based on nucleotide sequencing of the part of the HN gene showed that all BPIV3 isolates belonged to genotype C of BPIV3. They branched in one distinct cluster with three different branches, but these branches were very similar to each other (98.1% to 99.8%). Serbian BPIV3c isolates were most similar to the Chinese BPIV3c isolates SD0805, SD0809 and SD0835 (from 97.92% to 99.7%), and to South Korean (12Q061), Japanese (HS9) and American (TVMDL16 and TVMDL20) BPIV3c strains (from 97.1% to 98.8%), and distinct from American (TVMDL15and TVMDL17) and Australian (Q5592) BPI3V genotype B strains (only 79.9% to 82.3% similarity), as well as from the genotype A BPIV3 strains from different countries published in GenBank.

The present study aims to characterize ESBL-producing Escherichia coli isolated from healthy cattle and sheep in the Burdur province of Turkey. Fecal samples from a total of 200 cattle and 200 sheep were tested and ESBL-producing E. coli was isolated from 31 (15.5%) cattle and three (1.5%) sheep samples using the Clinical and Laboratory Standards Institute’s combined disk method. Among the ESBL gene classes detected by PCR, bla_CTX-M was the most frequent type, followed by the bla_TEM and bla_SHV families. ESBL-producing E. coli isolates showed co-resistance to multiple classes of antibiotics including aminoglycosides, phenicols, quinolones, folate pathway inhibitors and tetracyclines. The resistance rates were higher in the cattle isolates than in the sheep isolates. Phylogenetic grouping of the E. coli isolates indicated group A (particularly A1) was the predominant phylogenetic group (19/34, 55.9%), followed by groups B1 (9/34, 26.5%) and D (6/34, 17.6%); none of the isolates belonged to group B2. The study shows that ESBL-producing E. coli isolates exist in the intestinal flora of healthy cattle and sheep in the Burdur province of Turkey. This is the first report showing the emergence of CTX-M type ESBL-producing E. coli in sheep farms in Turkey

Data on the values of selected blood antioxidant parameters, i.e. total antioxidant capacity, glutathione peroxidase, and superoxide dismutase in healthy dogs, are lacking. There are no published accepted standard reference methods for their determination. The aim of this study was to determine the values of plasma total antioxidant capacity and the activities of whole blood glutathione peroxidase and erythrocyte superoxide dismutase in 30 healthy client-owned dogs (19 females, 11 males). The effect of age and sex on the measured antioxidant parameters was also investigated. Antioxidant parameters were determined with an automated biochemical analyser, using the commercially available Randox kits. No significant difference in age, weight, and antioxidant parameters was determined between females and males. A significant positive effect of age (p = 0.002, r² = 0.284) on superoxide dismutase activity was confirmed. There was no effect of sex on any of the antioxidant parameters measured. However, we observed a tendency of the effect of sex (p = 0.063, r² = 0.118), as well as age (p = 0.073, r² = 0.111), on the activity of glutathione peroxidase. Our results are in part comparable with the results of other studies in which the same types of methods and samples were used to determine antioxidant parameters. In conclusion, the sex and age of dogs should be taken into consideration when planning a study on antioxidant status parameters.

The aim of this study was to evaluate the reliability of multiple measurements with Lactate Scout portable analyzer in dogs during treadmill exercise. Ten Border collies were involved in the study and blood samples were taken before, three times during and twice after the treadmill exercise. Lactate concentration was measured in duplicate, by Scout portable analyzer and the reference biochemical analyzer in the laboratory, and the obtained values were compared. There was a high and positive correlation between these two methods (r=0.96, p=0.003). The Lactate Scout analyzer reveals a high degree of agreement with the laboratory method and therefore can be valid for use in research of veterinary sports medicine and emergency veterinary medicine where multiple measurements of lactate concentrations are often needed.

Malignant pilomatricoma is malignant follicular tumor with only matrical differentiation. Malignant pilomatricoma is rare in dogs. There is little information about sex, breed and age predisposition. There are a few reports of canine malignant pilomatricoma in middle to old age dogs. However, this neoplasm was resected from 1-year-old intact male miniature poodle. The neoplasm was found in the dorsal part of the neck. The mass was firm and protruded. On gross findings the size of the mass was 3×2×1.5cm. The mass was located in the deep dermis and subcutaneous layer. The mass was composed of several lobules of grey-white chalky material. Microscopically, the mass was composed of several large and small lobules. There were basophilic round to oval basaloid cells at the periphery of the lobules. The basophilic cells showed abrupt keratinization. Numerous ghost cells were observed in the center of the mass. The ghost cells had abundant eosinophilic cytoplasm without a nucleus. The basophilic basaloid cells showed numerous atypical mitotic figures. Cellularity was high and pleomorphism was remarkable. No lymphatic metastasis was observed. We reported a rare case of malignant pilomatricoma in a 1-year-old young dog.