Effects of Medetomidine-Butorphanol and Medetomidine-Buprenorphine on Oxidative Stress and Vital Parameters in Dogs Undergoing Ovariohysterectomy.

Oxidative stress (OS) is caused by an imbalance between the production of oxygen-containing free radicals and their elimination. General anesthesia increases the production of reactive oxygen species (ROS) and therefore causes oxidative stress. Our objective was to determine the effects of medetomidine-butorphanol (MEDBUT) and medetomidine-buprenorphine (MEDBUP) on oxidative stress and cardiorespiratory parameters in dogs undergoing ovariohysterectomy (OHE). Ten healthy female dogs were randomly assigned to two groups: the MEDBUT group (n = 5) received medetomidine and butorphanol, while the MEDBUP group (n = 5) received medetomidine and buprenorphine. OS was evaluated by measuring total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) during five different time points (from the administration of anesthetic drugs to 2 h after surgery). The observed vital cardiorespiratory parameters included heart rate (HR), respiratory rate (fR), noninvasive systolic (SAP) and diastolic (DAP) arterial blood pressures, oxygen saturation (SpO2), end-tidal CO2 (EtCO2), and body temperature (BT). Cardiorespiratory parameters were altered at a significantly greater degree in animals sedated with MEDBUT (p < 0.05). The administration of medetomidine-butorphanol was more likely to increase OS parameters, while medetomidine-buprenorphine showed decreased levels of oxidative stress throughout the study.

Association of mast cell density, microvascular density and endothelial area with clinicopathological parameters and prognosis in canine mammary gland carcinomas.

BACKGROUND: Mast cell density has been shown to have both enhancing and inhibiting effects on tumour progression and the ability to predict breast cancer behaviour in humans. However, prognostic results have been contradictory. Some previous studies suggested involvement of mast cells in the progression of canine mammary tumours. This study investigated total, intratumoural and peritumoural mast cell densities by Giemsa staining, and their association with clinicopathological parameters and the disease outcome of canine mammary tumours. In addition, since mast cells promote angiogenesis, the microvascular density and endothelial area were evaluated by CD31 immunostaining. RESULTS: Intratumoural mast cell density was associated with tumour size, lymph node involvement and tumour-infiltrating lymphocyte count, while peritumoural mast cell density was associated with grade. The endothelial area was associated with grade, mitotic index, tubular formation and proliferation index. Tumours with a high grade, high total intratumoural mast cell density and a larger endothelial area were associated with shorter disease-free survival. Intratumoural mast cell density and grade were found to be independent prognostic factors. CONCLUSIONS: These results suggest that intratumoural mast cell density and the endothelial area can be used to evaluate the aggressiveness of canine mammary carcinomas, while intratumoural mast cell density could be of use as an independent predictor of a prognosis of disease-free survival. Peritumoural mast cell density does not seem to influence tumour behaviour.

Awareness, Motivation, and Fear towards Canine Blood Donation-A Survey of Dog Owners in Lithuania.

The recruitment of canine blood donors remains a challenge, especially in countries where blood donation and veterinary medicine are still emerging medical fields. There are few previous studies that have discussed canine blood donation strategies, and the subject of fear and its influencing factors have not been investigated. The main purpose of our study was to investigate dog owners' awareness, motivation, and fear regarding canine blood donation in order to improve donor recruitment strategies. We created a six-page questionnaire and submitted it to dog owners (n = 207) in person. Two-thirds of the respondents (65.7%) were not aware that canine blood donation exists in Lithuania. We did not find any factors that would significantly affect the motivation of respondents toward donation. We found an association between the fear of the owner and the health status of the owned dog (p = 0.008), as well as if their animal had needed urgent care in the past (p = 0.031). The fact that some participants were blood donors themselves did not affect their motivation, but they were 19.76% less afraid of canine blood donation (p = 0.001), as were respondents who were aware of canine blood donation (p = 0.004). In conclusion, the recruitment strategy should focus on the management of fear toward canine blood donation and the education of clients, and donor welfare must remain a priority.

VEGF-B, VEGF-A, FLT-1, KDR, ERBB2, EGFR, GRB2, RAC1, CDH1 and HYAL-1 Genes Expression Analysis in Canine Mammary Gland Tumors and the Association with Tumor ClinicoPathological Parameters and Dog Breed Assessment.

Canine mammary gland tumors (CMTs) are one of the most prevalent cancers in dogs and a good model for human breast cancer (BC), however gene expression analysis of CMTs is scarce. Although divergence of genes expression has been found in BC of different human races, no such research of different dog's breeds has been done. The purpose of this study was to investigate expression of the VEGF-B, VEGF-A, FLT-1, KDR, ERBB2, EGFR, GRB2, RAC1, CDH1 and HYAL-1 genes of canine mammary carcinomas, compare the expression levels with clinicopathological parameters and analyze expression disparities between different breeds. Carcinomas and adjacent tissues were collected from female dogs to perform routine histopathology, immunohistochemistry (IHC) and quantitative real-time polymerase chain reaction (qRT-PCR). We found that VEGF-B and EGFR genes were overexpressed in the mammary gland carcinomas compared to adjacent tissue. VEGF-B gene expression had associations with different parameters (tumor size, grade, and absence of metastasis). Furthermore, differences in VEGF-B, FLT1, ERBB2, GRB2, RAC1, CDH1 and HYAL-1 genes expression have been found in different breed dogs (German Shepherd, Yorkshire Terrier) and mixed-breed dogs indicating that a dog's breed could determine a molecular difference, outcome of cancer and should be accounted as a confounding factor in the future gene expression research.

Bartonella Infections in Cats and Cat Fleas in Lithuania.

Bartonella are vector-borne parasitic bacteria that cause zoonotic infections in humans. One of the most common infections is cat-scratch disease caused by Bartonella henselae and Bartonella clarridgeiae. Cats are the major reservoir for these two species of bacteria, while cat fleas are vectors for the transmission of infection agents among cats. The aim of the present study was to investigate the presence of Bartonella infections in stray and pet cats and in cat fleas in Lithuania. Blood samples were taken from 163 cats presented in pet clinics and animal shelters. A total of 102 fleas representing two species, Ctenocephalides felis and Ctenocephalides canis, were collected from 12 owned cats that live both outdoors and indoors. Bartonella DNA in samples was detected using a nested PCR targeting the 16S-23S rRNA intergenic spacer (ITS) region. Bartonella DNA was detected in 4.9% (8/163) of the cats and 29.4% (30/102) of the fleas. Sequence analysis of the ITS region showed that the cats and fleas were infected with B. henselae, B. clarridgeiae and Bartonella sp., closely related to B. schoenbuchensis. This study is the first report on the prevalence and molecular characterization of Bartonella spp. in cats and cat fleas in Lithuania.

The influence of the duration of the preoperative time spent in the veterinary clinic without the owner on the psychogenic and oxidative stress in dogs.

The objective of this study was to evaluate the influence of the dog's long-term separation from its owner in the novel environment on the occurrence of psychogenic and oxidative stress. Group I dogs (n=9) were brought to the veterinary clinic and stayed in a kennel room for 12 hr before the surgery, and group II dogs (n=9) -for 10 min before the surgery. Physiological parameters (heart rate (HR) (beats/min) and respiratory rate (fR) (breaths/min)) were measured and blood sampling was done 12 hr before the surgery (T0) for group I dogs and 10 min before the surgery (T1) for both groups dogs. Oxidative stress index (OSI) was determined using spectrophotometer and Rel Assay Diagnostics kits by measuring TAS ant TOS in blood plasma. The cortisol level was measured using AIA-360 Automated Immunoassay Analyzer and ST AIA-pack Cortisol assays. Group I dogs' HR and fR were elevated at T0 and T1, and group II dogs'-at T1 compared to physiological range. OSI and cortisol levels in group I dogs was higher at T1 compared to T0 (P<0.05). There was no significant difference in cortisol level between group I at T0 and group II at T1 (P>0.05). It might be concluded that dogs' longer stay in the novel environment without the owner induced significant changes in OSI and cortisol level, which could lead to slow wound healing and the occurrence of systemic diseases.

Prevalence of anthelmintic resistance on Lithuanian sheep farms assessed by in vitro methods.

BACKGROUND: This study examines the prevalence of drug resistance in gastrointestinal nematodes to macrocyclic lactones (ML) and benzimidazoles (BZ) in Lithuanian sheep using sensitive and precise in vitro methods. The survey was conducted from August 2013 to November 2014. Thirty-three farms with sheep previously treated with BZ and ivermectin (IVM) were included in the study. On 12 farms where only BZ were used, egg hatch discrimination dose testing (EHDDT) was conducted to detect anthelmintic resistance (AR) to BZ. On eight farms where only ML were used, micro agar larval development testing (MALDT) was conducted to detect AR to ivermectin (IVM). On the remaining 13 farms, where both classes of drugs were used, EHDDT and MALDT were both applied to detect multidrug resistance to BZ and IVM. RESULTS: BZ-resistant gastrointestinal nematodes were found on all 25 farms with a previous history of BZ use. High levels of resistance (>40 % of hatching) were recorded on 36 % of these farms, and low levels (<20 % of hatching) on 40 % of farms. IVM-resistant populations were found on 13 out of 21 sheep farms using this drug. Of these 13 farms with AR to IVM, low levels of resistance (<30 % development) were recorded on 84.6 % of farms and high levels (>30 % development) on 15.4 % of farms. No resistance to IVM was recorded on 38.1 % of farms. Multi-drug resistance was detected on five farms out of 13 (38.5 %) using both classes of drugs. CONCLUSIONS: The present study demonstrates the existence of AR to BZ and ML on Lithuanian sheep farms thus confirming results in a previous in vivo study. Cases of multi-drug resistance were recorded in the present study and require further consideration. An appropriate strategy for anthelmintic treatment, measures to prevent gastrointestinal nematode infection and a better understanding of the management practices associated with resistance may slow down further development of AR.

Effect of active immunization against GnRH on testosterone concentration, libido and sperm quality in mature AI boars.

BACKGROUND: The aim of the present study was to investigate the efficacy of the Improvac on testosterone concentration in blood serum, sexual behavior and sperm quality in matured AI boars. A total of nine Danish Landrace AI boars were included in the analysis. METHODS: The trial period lasted for 15 weeks and was divided into four periods: Control period: three weeks before vaccination; Period I - four weeks after first vaccination; Period II - four weeks after second vaccination, Period III - four weeks after third vaccination. Blood and sperm samples were collected at weekly intervals. Freshly collected sperm samples were analyzed. RESULTS: Testosterone concentration correlated with libido (r = 0.531; p < 0.001), volume of ejaculate (r = 0.324; p < 0.001) and the percentage of morphologically normal spermatozoa (r = 0.207; p < 0.05). Testosterone concentration rised significantly (p < 0.05) in 5-6 week of trial, e. i. after the first dose of Improvac and after this peak the level of testosterone further progressively decreased (p < 0.05). CONCLUSIONS: Results from this study indicate that active immunization of sexually matured boars against GnRH has negative impact on testosterone concentration, sexual behavior, volume of ejaculate and total number of normal spermatozoa in ejaculate.

Assessment of sperm quality traits in relation to fertility in boar semen.

BACKGROUND: Several studies have been published where sperm plasma membrane integrity correlated to fertility. In this study we describe a simple fluorometer-based assay where we monitored the fluorescence intensity of artificially membrane-ruptured spermatozoa with a fixed time staining with fluorescent DNA dyes. METHODS: Membrane-impermeant fluorescent dyes Hoechst 33258 (H258) and propidium iodide (PI) were used to measure the fluorescence of the nucleus in artificially membrane ruptured spermatozoa and membrane-permeant dye Hoechst 33342 (H342) was used to measure fluorescence of intact spermatozoa. The concentration of spermatozoa in insemination doses varied from 31.2x10(6)/ml to 50x10(6)/ml and the average value was 35x10(6)/ml. Each boar was represented by three consecutive ejaculates, collected at weekly intervals. Nonreturn rate within 60 days of first insemination (NR%) and litter size (total number of piglets born) of multiparous farrowings were used as fertility measures. RESULTS: Sperm fluorescence intensity of H258 and H342, but not the fluorescence intensity of PI-stained spermatozoa correlated significantly with the litter size of multiparous farrowings, values being r=-0.68 (P<0.01) for H258, r=-0.69 (P<0.01) for H342 and r=-0.38, (P=0.11) for PI. CONCLUSIONS: The increase in fluorescence values of membrane-ruptured H258 and unruptured H342-stained spermatozoa in boar AI doses can be associated with smaller litter size after AI. This finding indicates that the fluorescence properties of the sperm nucleus could be used to select for AI doses with greater fertilizing potential.