Volume 68 (2018): Issue 1 (March 2018)

Although mycotoxins occur worldwide and represent a global public health threat, their prevalence and quantities in food and feed may vary due to geographic and climatic differences. Also, in accordance with climate change, outside temperatures that are anticipated to rise and rainfall patterns modify the usual mycotoxicological scheme transforms and unexpectedly extreme events happen in practice more often. Such weather conditions increase fungal occurrence and mycotoxin concentrations in crops. Consequently, the risk to human and animal health grows, and strategies to alleviate adverse effects become more complex. This also elevates economic losses. Therefore, the task of mycotoxin prediction has been put in front of the multidisciplinary scientific community recently, and a targeted prevention has become more important. This paper is a review of the latest achievements in this field prepared with the aim to summarize and integrate available data.

The main problems that cause unresponsiveness to an anti-neoplastic drug are the overexpression of drug resistant and anti-apoptotic proteins in tumor cells. In a rescue protocol we evaluated the ability of toceranib phosphate concurrent with lomustine (CCNU) or L-asparaginase and vincristine to decrease drug resistant and apoptotic proteins in relapsed and refractory canine lymphomas. The peripheral blood samples were collected before and after the rescue treatment from fourteen dogs that were refractory to cyclophosphamide-vincristine-prednisolone (COP) or COP-doxorubicin (CHOP) treatment and had recurrent multicentric lymphoma. The mRNA expression level of ABCB1, ABCG2, Bcl2 and Bax were determined by quantitative real-time PCR. The fold-change in ABCB1, ABCG2, Bcl2 and Bax mRNA levels were analyzed in correlation with the progression-free survival (PFS). After the rescue treatment, the ABCB1 and ABCG2 mRNA expression levels were 1.57- and 1.85-fold lower (p = 0.4 and p = 0.87), respectively, compared to pre-treatment. Bcl2/Bax ratio was numerically but not significantly decreased 1.02-fold (p = 0.74). The overall response rate of this protocol was 50% with a median PFS of 79 days (range 14-207 days). The low medians of relative expression levels of ABCB1, ABCG2 and Bcl2/Bax ratio group did not correlate with the clinical outcomes when compared to the high medians of relative expression levels, and likewise with the clinical stage, immunophenotype, histological grade and sub-stage. Therefore, the administration of a rescue drug with toceranib phosphate might be beneficial in refractory and relapsed canine lymphoma.

The aim of this study was to investigate the effects of total intravenous anaesthesia with propofol and anaesthesia induced with propofol and maintained with sevoflurane on oxidative stress parameters in dogs with early-stage myxomatous mitral valve degeneration (MMVD). Sixteen client-owned dogs with early stage MMVD that required periodontal treatment were included in the study. After induction with propofol, anaesthesia was maintained with propofol (group P) or sevoflurane (group PS). Blood samples for determination of vitamin E, superoxide dismutase, glutathione peroxidase and malondialdehyde were collected before premedication, 5 and 60 minutes and 6 hours after induction to anaesthesia. There were no significant differences between groups in any of the oxidative stress parameters at each sampling time. Compared to basal values, vitamin E concentration decreased significantly during anaesthesia in both groups and glutathione peroxidase activity increased 60 minutes after induction to anaesthesia in PS group. Anaesthesia with propofol or with propofol and sevoflurane did not have any significant impact on oxidative stress parameters in dogs with early stage MMVD. In terms of oxidative stress, both protocols may be equally safely used in dogs with early stage MMVD.

The Pain Trace™ device can detect changes in the skin's electrical potentials claimed to be associated with pain related alterations in the sympathetic and parasympathetic nervous system activity. Positive voltages represent the absence of major pain, whereas negative voltages represent moderate to severe pain. Unlike in humans and horses, no baseline skin potential recordings have been reported in dogs. In study Part 1 baseline skin potentials were recorded in healthy dogs and compared to readings obtained in human volunteers. In dogs, data were recorded with electrodes placed at three separate sites: neck, axilla, and thorax. In humans, data were collected from the palms. Readings over a 90-second period were averaged and comparisons between groups were performed using the Kruskal-Wallis test. All voltage recordings were positive. Readings in dogs had greater variability. Recordings from the thorax were more homogeneous, this being the reason why this site was chosen for study Part 2. No significant differences in recordings were noted between pain-free dogs and humans. The main hypothesis was that shifting from positive to negative skin potential voltages serves as an indicator of canine patients sensing moderate to severe pain. Therefore, we obtained preoperative readings from dogs with cranial cruciate ligament disease that were experiencing associated persistent pain, and compared these data with readings from pain-free dogs (thorax). In dogs undergoing surgery, all pre-surgery voltage readings were positive and thus no consistent relationship between skin potential recordings and pain perception could be established. Further investigation is needed to confirm any relationship between skin potential and pain severity in dogs.

To evaluate the efficiency of the probiotic strain Lactobacillus plantarum LS/07 in dextran sulphate sodium (DSS) induced acute colitis selected biochemical (activity of β-glucuronidase), microbiological (counts of lactobacilli and coliforms), and immunological (IL-6, IL-8, IL-13, NF-κB, MPO) parameters were assessed. Sprague-Dawley rats were divided into groups: Control, Acute colitis, and Probiotic. Acute colitis was induced using 5% DSS in drinking water for 7d. DSS induced an inflammatory process in the colonic tissue, increased the activity of β-glucuronidase (p<0.001), increased the counts of coliform bacteria and decreased lactobacilli counts (p<0.05), and activated the production of the measured parameters (NF-κB, MPO, IL-6, IL-8) except of IL-13. Lactobacillus plantarum LS/07 in the diet alleviated the DSS induced inflammatory process by inhibiting the production of IL-6, IL-8, the activities of NF-κB and MPO, and stimulated the production of IL-13. The probiotic reduced the activity of β-glucuronidase (p<0.05), increased lactobacilli counts and decreased coliform bacteria. These results indicate that dietary intake of Lactobacillus plantarum LS/07 suppressed the expression of markers playing an important role in the inflammatory process. We conclude that the anti-inflammatory properties of Lactobacillus plantarum LS/07 makes its suitable for the prevention or treatment of colitis.

A total of 131 blood samples and 175 spleen samples were collected from three cervid species: roe deer (Capreolus capreolus), red deer (Cervus elaphus) and fallow deer (Dama dama) inhabiting the continental part of Croatia. Serum samples were tested for antibodies against bovine herpesvirus 1, parainfluenza-3 virus, bluetongue virus, bovine respiratory syncytial virus, hepatitis E virus, bovine viral diarrhoea virus and enzootic bovine leukosis virus. The tested sera were negative for bovine viral diarrhoea virus, enzootic bovine leukosis virus, bluetongue virus, bovine respiratory syncytial virus and hepatitis E virus antybodies. The antibody prevalence in roe deer and red deer samples was 21.11% for bovine herpesvirus 1 and 75.55% for parainfluenza-3 virus. Sera from bovine herpesvirus 1 positive animals were subsequenty tested with comparative virus neutralization test and bovine herpesvirus 1 neutralising antibodies were found in 18 (out of 19) sera. In fallow deer, no antibodies against any of the viral pathogens were detected. All spleen samples tested for bovine viral diarrhoea virus and enzootic bovine leukosis virus came back negative, except for one red deer spleen sample found to be weakly diarrhoea virus-positive. Our findings provide the first information on the exposure of Croatia-inhabiting cervid species to viral pathogens, and could serve as valuable baseline data for future investigations regarding deer exposure to various pathogens and the distribution of diseases shared between wildlife and livestock. As of now, the epidemiology of these viruses in the Croatian cervid population has been only poorly understood, so that further research is recommended.

Canine parvovirus type-2 (CPV-2) is a DNA virus that causes a very common worldwide diffused infectious disease in dogs. Since its appearance, the CPV-2 evolved generating novel genetic and antigenic variants (CPV-2a/2b/2c) which are distributed throughout the world. In the present study, the frequency of CPV-2 in a canine population in Sicily (Italy) was investigated, using a polymerase chain reaction (PCR) for a fragment of the VP2 gene. Out of a total of 673 samples from 370 dogs, submitted to the laboratory from July 2009 to August 2015, 265 samples (39.38%) were positive and these were further analyzed by restriction fragment length polymorphism (RFLP) and DNA sequence analysis. A high prevalence of the CPV-2c variant (79.56% of CPV-2 positive dogs) was observed in this survey, underlining that this type of CPV field isolate is prevalent circulating in Sicily. Sequence and phylogenetic analysis showed a close relationship with CPV-2a and CPV-2c strains from Europe and non-European countries. Considering that CPV-2c is reaching a worldwide distribution and that this variant is also affecting vaccinated dogs, efforts should be made towards the development of new CPV vaccines.

Antibiotic therapy of staphylococcal mastitis is characterized by significantly lower cure rates compared to infections caused by other microorganisms. Thus, it is necessary to search for new, alternative, non-antibiotic agents that are effective in the eradication of these bacteria. The aim of our research was to investigate the antimicrobial, especially anti-staphylococcal potential of a large collection (n=36) of essential oils (EOs). Investigation of the antimicrobial activity of tested oils was determined by using a serial, twofold dilution method in 96-wells microtiter plates under conditions recommended by the Clinical and Laboratory Standards Institute (CLSI). The preliminary analysis revealed that six oils, namely: Manuka, Thyme, Geranium, Cedar, Cinnamon (from bark) and Patchouli exhibited the highest activity against reference strains of bacteria. Significant anti-staphylococcal potential of these oils has been also confirmed for a group of 18 Staphylococcus aureus, 8 Staphylococcus epidermidis and 5 Staphylococcus xylosus strains isolated from cases of bovine mastitis. Especially high activity was observed for Cedar, Patchouli, Thyme and Manuka oils. The MIC (Minimal Inhibitory Concentration) values for Patchouli oil were in the concentrations range of 0.01 to 0.313% (v/v). The three other oils inhibited the growth of staphylococci isolated from mastitis in the concentrations range of 0.01 to 0.625% (v/v). Oils isolated from Cinnamomum cassia and Pelargonium graveolens revealed a bit lower, but still satisfactory activity (MIC values in the concentrations range of 0.02 to 1.25% (v/v) and from 0.078 to 1.25% (v/v), respectively). In many cases a slightly higher concentration of oils was required to obtain the bactericidal effect in comparison to growth inhibition. The time – kill kinetic assay revealed that the bactericidal effect was achieved after two hours incubation of the reference strain S. aureus PCM 2051 cells with Thyme oil at concentration equal to 2xMIC (1.25% (v/v)) or MIC (0.625% (v/v)). A slightly lower activity was observed in the case of Cinnamon oil, the bactericidal effect was achieved after 8 hours of incubation. The results of our research clearly indicate that some essential oils exhibit a promising antimicrobial activity and can be considered as alternative antistaphylococcal agents.

Ketamine and magnesium can interact in additive, supra-additive and antagonistic manners in analgesia or anesthesia. Ketamine is a non-competitive NMDA receptor antagonist. Magnesium is an endogenous non-competitive NMDA antagonist that causes anion channel blockade in a dose-dependent manner. It has been established that ketamine and magnesium interact synergistically in the tail-immersion test in rats.

To determine the role of serotonergic, GABAergic and noradrenergic systems in analgesia induced by the ketamine-magnesium sulfate combination.

Experiments were performed on male Wistar albino rats (200-250 g). Antinociception was evaluated by the tail-immersion test.

Methysergide (0.5 and 1 mg/kg, sc) administered alone did not affect nociception in rats. Methysergide (0.5 and 1 mg/kg, sc) antagonized the antinociceptive effect of the ketamine (5 mg/kg)-magnesium sulfate (5mg/kg) combination. Bicuculline (0.5 and 1 mg/kg, sc) given alone did not change the threshold to thermal stimuli in rats. Bicuculline (0.5 and 1 mg/kg, sc) antagonized the antinociceptive effect of the ketamine (5 mg/kg)-magnesium sulfate (5 mg/kg) combination. Yohimbine (0.5, 1 and 3 mg/kg, sc) applied alone did not change nociception. Yohimbine at a dose of 0.5 mg/kg did not influence the effect of ketamine (5 mg/kg)-magnesium sulfate (5 mg/kg), while yohimbine at doses of 1 and 3 mg/kg antagonized the antinociceptive effect of this combination.

Serotonergic, noradrenergic and GABAergic systems participate, at least in part, in the antinociceptive effect of the ketamine-magnesium sulfate combination in acute pain in rats.

Cementoblastoma is a rare, benign, expansive and slow-growing, mesenchymal odontogenic tumor. An eight year-old, Purebred Spanish Horse stallion with clinical history of chronic, refractory to treatment unilateral rhinitis and radiologic evidence of a radiopaque mass at the second premolar tooth root underwent intraoral surgery for tooth and mass extraction. The owner did not want to bear the cost of the histopathologic study and the samples were left unattended in a sterile recipient without a fixative solution for six years. Grossly the mass showed a hard consistence, greyish-whitish color and an irregular surface. Histologically, the mass consisted of an abundant deposit of eosinophilic cementum-like organic matrix with frequent basophilic reversal lines and heterogeneous distribution of empty lacunae. Surrounded by this organic matrix there were numerous fungal hyphae positive for Aspergillus fumigatus immunostaining. The final diagnosis was maxillary cementoblastoma with secondary aspergillosis. High-quality information has been obtained from long-time non-fixed tissues.