The Efficiency of Synthetic Sex Pheromones in Sexual Arousal Stimulation in Domestic Dogs

The aim of this study was to evaluate the suitability of synthetic bitch sex pheromones (Eau’ De Estrus®, Synbiotics USA) for the stimulation of the reproductive reflexes in adult male dogs. In experiment I: anoestral bitches were applied synthetic (n=6) or natural (n=6) pheromones and their attractiveness was compared to the attractiveness of bitches in natural oestrus, In experiment II: swabs socked in natural estrual discharge or Eau D’Estrus were presented to male dogs and behavioral arousal (Exp. II a) and changes in blood flow in the penile artery reflecting erection (Exp. II b) were recorded. In experiment III the time dedicated for sniffing in experimental dogs was analyzed in relation to the type of attractant presented (natural estrual discharge or Eau D’Estrus). In all three experiments we observed the signs of sexual arousal, increased blood flow in the penile artery and increased time of sniffing only with natural pheromones. No signs of excitation or changes in penis perfusion were noted after the use of artificial pheromones. Our results did not support the suitability of Eau D’Estrus for male dog sexual stimulation.

Plasma profile of cimicoxib in sheep after oral administration at two different rates

Sheep are often subjected to painful procedures and thus they need to be treated with analgesics. Nevertheless, knowledges about pharmacokinetic features of these drugs in this species are poor. The aim of this study was to evaluate plasma behaviour of cimicoxib in sheep after a single oral administration at two different dose rates (4 and 6 mg/kg). Maximum plasma concentrations of cimicoxib were equal to 273.78 (median value; range 189.00-567.32) and 565.01 (range 308.27-822.59) ng/mL after treatment with 4 and 6 mg/kg, respectively. The time of maximum concentration (Tmax) was achieved between 4 and 10 hours following treatment at the lower dose, and between 6 and 10 hours after the administration of the higher dose, with one sheep achieving the concentration peak at 0.75 hours. The slow absorption and the great individual variability in plasma concentration, probably due to ruminal effects, suggest that cimicoxib is not suitable for oral treatment in sheep.

Differentiation of infectious bronchitis virus vaccine strains Ma5 and 4/91 by TaqMan real-time PCR

The aim of this study was to develop rapid molecular assays for differentiating vaccine strains Ma5 and 4/91 of the infectious bronchitis virus (IBV). Specific primers and probes for S1 and N genes were designed based on the nucleotide sequences of both vaccine strains. Cross-reactivity was not observed. Assay sensitivity was 2.373 × 103 copies of the Ma5 strain, and 3.852 x 103 copies of the 4/91 strain. Samples belonging to a known genotype demonstrated that the designed assays supported rapid and sensitive detection of Ma5 and 4/91 vaccine strains of IBV.

First report of Feline Calicivirus (FCV) infection in stray cats in northeast China

To improve our understanding of Feline calicivirus (FCV) infection in cats in Northeast China, 1584 serum samples from 974 domestic cats and 610 stray cats were collected between 2012 and 2015. The samples were tested for FCV antibodies using a commercially available ELISA kit. The results revealed an overall seroprevalence of 37.56% (595/1584), a seroprevalence in domestic cats of 32.85% (320/974) and a seroprevalence in stray cats of 45.08% (275/610). Risk factor analysis indicated that species was the only risk factor for the presence of FCV (OR=1.678, 95% CI=1.362-2.066, P<0.001); age, season, region and gender were not risk factors. This is the first report of FCV infection in stray cats in China, and the results of this study can aid in FCV infection control in the felidae family.

Evaluation of usefulness of a commercial agarose gel electrophoresis kit (QuickGel SP) for bovine serum protein electrophoresis

The aim of this study was to show the usefulness of a commercial agarose gel electrophoresis (AGE) kit (QuickGel SP) for separating bovine serum protein fractions in comparison with conventional cellulose acetate electrophoresis (CAE). Serum protein bands were verified using five reference reagents corresponding to albumin and α1-, β1-, β2-, and γ-globulins. AGE clearly revealed six separated fractions of albumin and α1-, α2-, β1-, β2-, and γ-globulin fractions in 100% and 77.8% in serum samples of dairy cows from the healthy (n=27) and diseased groups (n=27), respectively. The α1- and α2-globulins were not separated by CAE in 14.8% and 96.3% of the samples from the healthy and diseased groups, respectively, whereas β2- and γ-globulin were not separated by CAE in 96.3% and 100% of the samples from the healthy and diseased groups, respectively. More than 94% of the points for the α-globulin fractions (α1- and α2-globulins), the β-γ-globulin fractions (β1-, β2-, and γ-globulins), and the albumin/globulin ratio between AGE and CAE were within agreement on the Bland-Altman plots. However, the mean biases were not near zero in the albumin and β-γ-globulin fractions. These results suggest that the high-resolution commercial AGE kit can be utilized to separate bovine serum protein fractions.

An evaluation of the usefulness of invasive and non-invasive methods used to diagnose Helicobacter spp. infections in dogs

The aim of this study was to assess the suitability of invasive and non-invasive methods used to diagnose Helicobacter spp. in the stomachs of dogs. The study was carried out on 30 dogs of both sexes and different breeds, between one and 15 years old. A histopathologic examination, a microbiological culture, a rapid urease test, a direct bacteriological preparation and a nested PCR assay were carried out. Gastric Helicobacter spp. was identified in gastric biopsy specimens from 16 (53.3%) dogs using direct bacteriological preparation, in four (13.3%) dogs based on a culture, in 23 (76.6%) dogs using the rapid urease test and in 21 (70,0%) dogs based on a histopathological assessment of the biopsy specimens. The nested PCR of the gastric biopsy specimens revealed gastric Helicobacter spp. in all the dogs (100%). A saliva PCR assay revealed gastric Helicobacter spp. in 23 (76.6%) dogs, while stool PCR revealed the bacterium in seven (23.3%) dogs. We found that invasive methods were more accurate than non-invasive methods in detecting a Helicobacter spp. infection in dogs. In addition, the nested PCR method used to evaluate the gastric mucosal biopsy specimens was the most accurate test for detecting Helicobacter spp. It was further found that the PCR-based saliva assay was the best non-invasive method for detecting Helicobacter spp. However, taking into consideration that most of the diagnostic methods used to detect this bacterium have drawbacks, at least two diagnostic methods should be used to detect Helicobacter spp. as is done in human medicine.

Detection of Yersinia Enterocolitica Species in Pig Tonsils and Raw Pork Meat by the Real-Time Pcr and Culture Methods

The aim of the present study was to establish a rapid and accurate real-time PCR method to detect pathogenic Yersinia enterocolitica in pork. Yersinia enterocolitica is considered to be a crucial zoonosis, which can provoke diseases both in humans and animals. The classical culture methods designated to detect Y. enterocolitica species in food matrices are often very time-consuming. The chromosomal locus _tag CH49_3099 gene, that appears in pathogenic Y. enterocolitica strains, was applied as DNA target for the 5’ nuclease PCR protocol. The probe was labelled at the 5’ end with the fluorescent reporter dye (FAM) and at the 3’ end with the quencher dye (TAMRA). The real-time PCR cycling parameters included 41 cycles. A Ct value which reached a value higher than 40 constituted a negative result. The developed for the needs of this study qualitative real-time PCR method appeared to give very specific and reliable results. The detection rate of locus _tag CH49_3099 - positive Y. enterocolitica in 150 pig tonsils was 85 % and 32 % with PCR and culture methods, respectively. Both the Real-time PCR results and culture method results were obtained from material that was enriched during overnight incubation. The subject of the study were also raw pork meat samples. Among 80 samples examined, 7 ones were positive when real-time PCR was applied, and 6 ones were positive when classical culture method was applied. The application of molecular techniques based on the analysis of DNA sequences such as the Real-time PCR enables to detect this pathogenic bacteria very rapidly and with higher specificity, sensitivity and reliability in comparison to classical culture methods.

Adenocarcinoma of the posterior segment of the gastrointestinal tract in dogs – clinical, endoscopic, histopathological and immunohistochemical findings

Of all the tumours in dogs, three percent are located in the intestines, and 36-60% of those tumours affect the large intestine. Adenocarcinomas of the intestines account for 20-35% of the gastrointestinal tumours and for almost 60% of the large intestine tumours. The aim of the study was to analyze clinical disorders and endoscopic, histopathological and immunohistochemical changes in colorectal adenocarcinomas in dogs with the use of the E-cadherin, β-catenin, cytokeratin 20 (CK20), Ki-67 and minichromosome maintenance 3 (MCM-3). The study comprised 11 dogs of both genders and of different breeds diagnosed with adenocarcinoma of the large intestine. They were from 4 to 11 years old. The large intestine adenocarcinoma was diagnosed in all the patients. 72.7% cases were diagnosed with a rectal adenocarcinoma, and 27.3% were found to have a colonic adenocarcinoma. All the studied proteins were expressed at different levels and, together with the histological findings, indicated different levels of malignancy (G). The statistical analysis revealed no statistically significant differences between the expression of E-cadherin and β-catenin in the studied tissues (p=0.79) and between the expression of Ki-67 andMCM-3 (p=0.39). A strong positive correlation was found between the expression of E-cadherin and β-catenin (r=0.86; p<0.05). The diagnosis of adenocarcinomas of the large intestine may be facilitated by the introduction of immunohistochemical studies using appropriate cell markers. They may also aid in the accurate evaluation of the biological character of the tumours, their origin, the connections between tumour cells and the mitotic index. That, in turn, may help determine the malignancy and the choice of treatment.

Interactions between plant-derived oestrogenic substances and the mycoestrogen zearalenone in a bioassay with MCF-7 cells

Human and animal diets may contain several non-steroidal oestrogenic compounds which originate either from plants (phytoestrogens) or from fungi that infect plants (mycoestrogens such as zearalenone (ZEN)). Phytoestrogens may compete with ZEN in binding to the oestrogen receptor β and thereby may counteract the oestrogenic activity of ZEN. Using a modified version of the E-screen assay, plant-derived oestrogenic substances were tested for their proliferative or anti-proliferative effect on oestrogen-dependent MCF-7 cells. The samples were additionally tested for their ability to influence the oestrogenic activity of ZEN (1 μM). Among the individual substances tested, 8-prenylnaringenin had the strongest effect, as cell proliferation was increased by 78% at the lowest concentration (0.23 μM), and by 167% at the highest concentration (29.4 μM). Coumestrol (5.83 μM) increased cell proliferation by 39%, and genistein (370 μM) by 61%, respectively. Xanthohumol and enterolactone did not stimulate cell proliferation significantly. In the co-incubation experiments with ZEN, none of the single substances was able to decrease the oestrogenic activity of ZEN. Only for 8-prenylnaringenin (14.7 and 29.4 μM) was a trend towards an increase in the ZEN-induced cell proliferation up to 72% observed. In conclusion, with the exception of 8-prenylnaringenin, no substantial interaction between phytoestrogens and the mycotoxin ZEN could be detected using a bioassays with MCF-7 cells.

Cardiac disorders in dogs infected with Babesia canis

The aim of this study was to assess cardiac disorders in dogs infected with B. canis. The study included 50 dogs with babesiosis and 20 healthy control animals. All the animals had haematological tests, ECG, echocardiography and serum troponin I and CK-MB levels checked. The haematology in the group of dogs with babesiosis confirmed thrombocytopaenia in 100% of dogs, decreased haematocrit in 52% and anaemia in 46%. The most common abnormalities in ECG and echocardiography in dogs infected with protozoa included: change in appearance and/or amplitude of the T-wave (34%), increased fractional shortening (24%), an increased sinus rhythm (14%) and heart axis deviation (10%). In 19 of the 50 dogs with babesiosis, the level of serum troponin I was elevated. In 2 dogs that died from babesiosis, the troponin level I was very high. The ECG confirmed sinus tachycardia and interpolated ventricular beat in these animals. In all dogs with babesiosis that were used in the study, the serum CK-MB was high or very high and was within limits of 23.17 U/L - 369.62 U/L. The highest kinase concentration (367.33 U/L and 369.62 U/L) was observed in dogs that died due to the disease. The presented results prove that cardiac changes are common in canine babesiosis, but that most changes are nonspecific and appear to have little clinical significance. Cardiovascular assessment should be based on the assessment of the level of troponin I and CK-MB in the serum of sick animals. High concentrations of these factors might be indicators of poor prognosis.