Volume 67 (2017): Issue 1 (March 2017)

The study of cell proliferation is a useful tool in the fields of toxicology, pathophysiology and pharmacology. Cell proliferation and its degree can be evaluated using 5-bromo-2′-deoxyuridine which is incorporated into the newly synthesized DNA. The aim of this study was the optimization of subcutaneous application of 5-bromo-2′-deoxyuridine implantation for continuous and persistent marking of proliferating cells in the rat forestomach. 3-tert-Butyl-4-hydroxyanisole was used as the agent that ensures cell proliferation. In order to determine the optimal dose for proliferating cells labeling, 5-bromo-2′-deoxyuridine doses of 50 mg, 100 mg, 200 mg or 350 mg were implemented 2 days prior to sacrifice by flat-faced cylindrical matrices. Immunohistochemical analysis using 5-bromo-2′-deoxyuridine in situ detection kit was performed for the detection of 5-bromo-2′-deoxyuridine labeled cells. The results showed that for adult rats, the optimum 5-bromo-2′-deoxyuridine dose is 200 mg per animal for subcutaneous application. The here described manner of 5-bromo-2′-deoxyuridine in vivo labeling provides a simple, efficient, and reliable method for cell labeling, and at the same minimizes stress to animals.

Canid herpesvirus 1 (CaHV-1) is a member of the canine infectious respiratory disease complex (CIRDC). The outcome of CaHV-1 infection can be occasionally fatal. So far, no information on CaHV-1 circulation in Thailand has been reported resulting in a lack of preventive strategies. In this study, nasal (NS) and oropharyngeal (OS) swabs were collected from 100 live dogs with respiratory distress. Among them, 23 pleural effusions were aspirated. A panel of CIRDC-associated viruses was screened by (RT)-PCR, including CaHV-1, CIV, CPIV, CDV, CRCoV and CAdV-2, for all collected samples. The CaHV-1 was detected in 32 dogs. Additionally, CaHV-1 was consistently detected in six pleural effusions. Most CaHV-1 infected dogs were over 5 years of age (43.8%) and expressed a mild nasal discharge. Pathological results of four three-month-old puppies, naturally moribund from respiratory disease, revealed a severe multifocal necrotic-hemorrhagic disease in several organs without pathognomonic inclusion bodies. They were only found to be CaHV-1 positive by PCR. Phylogenetic analysis demonstrated concordant results of CaHV-1 circulation in Thailand. Although mostly found as a co-infection with other CIRDC viruses (68.8%) it also occurred alone. Therefore, rapid ante-mortem diagnosis might facilitate the investigation of unclassical CaHV-1 infection, which is fatal in neonates and causes illness in annually core-vaccinated adults.

The aim of this study was to evaluate the dynamic changes of serum lipid and lipoprotein profiles in 6 newborn calves during the first five days of life. From each calve blood sampling was performed daily starting from day 1 (after colostrum intake) until day 5 of life. Blood samples collected from each animal were tested for serum total lipids, phospholipids, non-esterified fatty acids (NEFAs), triglycerides, very low density lipoproteins (VLDLs), total cholesterol (Total-Chol), high density lipoproteins (HDLs) and low density lipoproteins (LDLs). One-way repeated measures analysis of variance (ANOVA) was applied to determine the effect of days of life on the studied parameters in calves. A statistically significant effect of days of life was found on all serum lipid and lipoprotein indices measured in calves with the exception of NEFAs that showed unchanged values throughout the monitoring period. The changes observed in calves during the early postnatal period are most likely due to the transition in energy sources, from a maternal nutrient supply comprising mainly carbohydrates and amino acids to the colostrum and milk diet rich in fat.

The aim of this present work was to examine statistical errors in published veterinary science articles. A total of 204 published articles (SCI or SCI-Exp) were used in this study. The articles were chosen from among those indexed in PubMed database between the years 2010 and 2014, inclusive. A total of 199 articles had at least one statistical error. The most frequently encountered statistical error among the articles published in journals indexed in SCI and in SCI-E was “errors in summarizing data”. No statistical error was found in 2.45% (n=5) of 204 (SCI: 0.98% (1/102), SCI-E: 3.92% (4/102)) articles. To reduce and prevent statistical errors in publications, the researchers must have a basic knowledge of statistics and during the study process they must consult field experts. While reviewing, the reviewers have to redirect the publications to statistical editors when needed and most importantly during the process of editing, the editors have to direct the publications to a statistics reviewer.

The aim of this study was to assess the genetic variations and relationships across evolutionary related cervid species in order to estimate the genetic diversity of the Red deer population that inhabits the forest area in the south-western part of Slovakia. The study was based on the application of cross-species SNP genotyping. The genomic data were obtained from a total of 86 individuals representing six genera (Axis, Dama, Cervus, Alces, Rangifer, and Odocoileus) using Illumina BovineSNP50 BeadChip. From 38.85% of successfully genotyped loci up to 1,532 SNPs showed polymorphism and were informative for subsequent analyses of the diversity and interspecific genetic relationships. Generally, a good level of observed heterozygosity was found across all species. The value of F_IS (0.23±0.13) signalised the increase of a homozygous proportion within them. The application of molecular variance analysis to the hierarchical population structure showed that most of the variation was conserved within separate species (96%). The performed diversity analysis of Slovak Red deer population and comparative analysis of their phylogenic relationships among subspecies from genus Cervus did not identify a remarkable loss of genetic variability. Also, were not identified any degree of admixture that could be due to the historical background of deer farming in Slovakia or reintroduction and hybridisation by other species from genus Cervus (C. canadensis, and C. nippon) which are the major risk of loss of autochthonous Red deer populations in many areas of Central Europe. The analysis of individual’s ancestry showed consistent results with patterns of evaluated group differentiations which means low migration rates among all species.

Insulin resistance is a phenomenon which accompanies the ongoing metabolic adaptation in cows during early lactation. The aim of our study was to determine the linear correlations of HOMA (Homeostatic Model Assessment), QUICKI (Quantitative Insulin Sensitivity Check Index) and RQUICKI (Revised Quantitative Insulin Sensitivity Check Index) indexes of insulin resistance with the metabolic status of cows (concentration of hormones, metabolites and body condition score). The experiment included 40 Holstein-Frisian cows in the first week after calving. Indexes of insulin resistance valued: 18.68±5.43 (HOMA), 0.39±0.06 (QUICKI) and 0.45±0.06 (RQUICKI). Linear correlations were examined by testing the coefficient of correlation (r), determination (r²,%) and regression parameter beta (b) in linear equation. A negative correlation was found between HOMA and IGF-I (insulin growth factor I) (r=−0.51, r²=25.0, b=−1.1257, p<0.01). HOMA showed a positive correlation with BHB (betahidroxybutyrate) (r=0.48, r²=23.2, b=0.0234, p<0.01). A positive correlation was found between QUICKI and IGF-I (r=0.30, r²=10.0 b=46.7900, p<0.05) and cholesterol (r=0.44, r²=18.3, b=1.9021, p<0.01). In contrast, QUICKI and BHB (r=0.51, r²=27.1, b=−1.7241, p<0.01), just like QUICKI and BCS (r=0.46, r²=20.9, b=−2.424, p<0.01), showed a negative correlation. RQUICKI showed positive correlations with IGF-I (r=0.48, r²=22.8, b=28.1230, p<0.01), T4 (r=0.47, r²=22.1, b=87.142, p<0.01) and triglycerides (r=0.36, r²=13, b=0.0407, p<0.05) but negative correlations with cortisol (r=−0.36, r²=13.0, b=−9.0332, p<0.05), STH (somatotropic hormone) (r=−0.42, r²=17.3, b=−5.4976, p<0.01), BHB (r=−0.62, r²=38.3, b=−1.1872, p<0.01), total bilirubin (r=−0.58, r²=33.7, b=−7.131, p<0.01) and BCS (body condition score) (r=−0.6, r²=36.4, b=−1.8347, p<0.01). In conclusion, indexes of insulin resistance may be used to evaluate the metabolic status of cows in early lactation. RQUICKI might be the most appropriate predictor of metabolic status due to its linear relationship with most of the parameters included in homeorhetic process.

This paper presents the results of the research on renal blood vessels of the mink. The main goal of this research is to provide a unique insight in renal vessel constellation and thereby contributes to the amount and quality of the available data on the cardiovascular system of the mink, since the data found in the available literature regarding this specific field is rather poorly documented. Blood vessels of the mink kidney were studied by dissection and corrosion injection technique. The renal artery and its branches, and the lobar, interlobar and interlobular arteries are terminal arteries and they do not anastomose between each other. The arterial system is characterized by two independent vascular areas, supplied by the dorsal and ventral lobar artery. Lobar arteries divide the kidney horizontally into a dorsal and ventral half, while lobar veins divide it vertically into the cranial and caudal half. The renal vein is formed from the cranial and caudal lobar vein. The obtained results are compared with relevant literature findings on renal blood supply of a dog and other sympatric carnivores.

The aim of this study was to investigate the morphology of Demodex canis and Demodex sp. cornei found in six dogs with canine demodicosis. A deep skin scraping technique was used for Demodex mite detection. Measurement data of 52 adult D. canis mites (26 females, 25 males and one specimen whose sex could not be determined) and 39 adult Demodex sp. cornei mites (22 females, 14 males and three specimens whose sex could not be determined) were reported. The correlation between body size of both Demodex species were estimated by the Student’s t-test. There was a significant correlation between short-tail and long-tail forms and total body length and length of the podosoma and opisthosoma (p<0.05). A significant difference was not found between the length of the gnathosoma and short-tail and long-tail forms (p>0.05). Demodex sp. cornei and D. canis, found in dogs from Vladivostok, were smaller than species from other countries. However, the present data did not significantly differ from other studies with D. canis and Demodex sp. cornei descriptions.

Induced prenatal hypothyroidism in rat pups leads to accelerated primordial follicle assembly and premature follicular atresia with ovary failure. This work investigates the influence of maternal hypothyroidism induced with 6-n-propyl-2-thyouracil (PTU) on the number and morphology of oogonia/oocytes in newborn rat pups with light and transmission electron microscopy. Expression of apoptosis and autophagy markers in oogonia/oocytes were examined using immunohistochemistry. Hypothyroid newborn pups had a decreased number of mitotic and resting oogonia, while the number of altered oogonia/oocytes was increased. Ultrastructural observations revealed the increased presence of degenerated pachytene oocytes (Z-cells) and oogonia/oocytes undergoing autophagy, apoptosis and combined apoptosis and autophagy, in this group. The most abundant altered oogonia/oocytes in the hypothyroid group were those with morphological features of advanced autophagy and Z-cells. The percentage of TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) positive oogonia/oocytes was significantly lower in the hypothyroid group. No significant difference was recorded in the expression of caspase-3, ATG7 and LC3 possibly reflecting that these proteins were not involved in the oogonia/oocyte alteration process during prenatal rat hypothyroidism. The obtained results indicate that developmental hypothyroidism in the offspring enhances the number of Z-cells and oogonia/oocytes altered with the advanced process of autophagy.

A nutritional placebo-controlled study was performed in Wistar rats in order to investigate the effects of 5-weeks aronia juice consumption towards fatty acid (FA) composition of phospholipids in the plasma and liver, as well as plasma glucose (Glu) and cholesterol levels. The animals were divided into 3 groups of 8 animals each, and randomized to receive either the full polyphenol dose of Aronia melanocarpa juice (AMJ), 4 times less polyphenol dose (¼-AMJ) or polyphenol-lacking placebo beverage (PLB). Each group of 8 male adult Wistar rats received the liquid ad libitum. AMJ decreased the levels of low-density lipoprotein (LDL) (P < 0.05) vs. PLB. AMJ increased dihomo-γ-linoleic acid (DGLA, 20:3n-6) (P < 0.05) and decreased arachidonic acid content (AA, 20:4n-6) (P < 0.05) vs. PLB in liver phospholipids. AMJ significantly increased monounsaturated fatty acids (MUFA) levels both in the liver (P < 0.05) and plasma (P < 0.05). Both aronia juice doses elevated the levels of beneficial n-3 polyunsaturated fatty acids (PUFA) in the plasma and liver. There was a dose-dependent, significant increase (P < 0.001) in cis-vaccenic acid (VA, 18:1n-7) in phospholipids in the plasma and liver. Our results indicate favorable effects of aronia juice intake on lipid parameters in Wistar rats. These findings suggest the potential of aronia dietary intake in cardiometabolic diseases primary prevention strategies in the human population.

Spontaneous mammary tumors are very common in bitches. The involvement of oxidative stress and the function of antioxidant enzymes in cancerogenesis have been studied in depth in human medicine, while data in veterinary medicine are still fragmentary. The main aim of this study was to evaluate the activity and the expression of superoxide dismutases (Cu-ZnSOD and MnSOD) and the activity of catalase (CAT) in canine mammary tumors in comparison with the adjacent healthy tissue. Six female dogs (mean age 10.4 years) were included in this study. After surgery, fresh tumor and healthy tissue samples were immediately frozen in dry ice and stored at −80°C for biochemical analyses, while the remaining parts were used for histopathological analysis. Enzyme activity was measured by spectrophotometric assays and protein expression by western blotting. In canine mammary tumors, Cu-ZnSOD activity and expression increased significantly compared with healthy control tissues (p=0.03). MnSOD showed a significantly lower activity in tumoral tissues at stage 2 (p<0.05), while a significant increase of expression was measured in tumors. CAT activity was significantly higher in healthy tissues respect to tumors (p=0.015). These variations of antioxidant enzymes activities and expression could be related to an increase of oxidative stress in breast cancer tissues and could be considered as biomarker candidates for neoplastic transformation.

CRP and WBC are the most widely used markers of inflammation in veterinary clinical diagnosis. Also, PCT is a specific APP marker of bacterial diseases in humans. This study evaluated the levels of PCT, CRP and WBC during postoperative monitoring of bitches undergoing ovariohysterectomy. Thirteen adult clinically healthy female crossbreed dogs aged between 7-24 months were used in the study. Ovariohysterectomy was performed via medial laparotomy using routine procedures. Blood samples were collected from the cephalic vein at 1, 4 and 7 days after surgery (post-op period). Mean serum concentration of PCT, CRP and WBC levels were increased after the surgery. CRP concentrations and WBC counts were increased significantly (p<0.001) on the first day after the operation and decreased to basal values. However, the PCT rise was not significant. CRP and WBC levels increased rapidly and decreased to normal values in dogs with ovariohysterectomy. CRP and WBC may be of help to determine the possible postoperative complications. Besides, aseptic surgical trauma did not affect PCT levels in dogs. Future studies are needed on canine PCT response focusing on specific bacterial infections.