Volume 63 (2019): Issue 2 (June 2019)

Introduction: Bovine parainfluenza virus-3 (BPIV3) and bovine respiratory syncytial virus (BRSV) are the cause of respiratory disease in cattle worldwide. With other pathogens, they cause bovine respiratory disease complex (BRDC) in ruminants. The aim of the study was the detection and molecular characterisation of BPIV3 and BRSV from nasal swabs and lung samples of cows in and around the Erzurum region of eastern Turkey. Material and Methods: In total, 155 samples were collected. Of animals used in the study 92 were males and 63 females. The age of the animals was between 9 months and 5 years, mean 1.4 years. Most males were in the fattening period and being raised in open sheds; females were in the lactating period and kept in free stall barns. All samples were tested for the presence of viral genes using RT-PCR. Gene-specific primers in a molecular method (RT-PCR) identified BRSV (fusion gene) and BPIV3 (matrix gene) strains at the genus level. Results: RNA from BRSV and BPIV3 was detected in two (1.29%) and three (1.93%) samples, respectively, one of each of which was sequenced and the sequences were aligned with reference virus strains. Phylogenetic analyses clustered the strains in genotype C/BPIV3 and subgroup III/BRSV. Conclusion: The results indicate that BRSV and BPIV3 contribute to bovine respiratory disease cases in Turkey. This is the first report on their detection and molecular characterisation in ruminants in Turkey.

Introduction: Numerous mutations in the bovine tumour necrosis factor receptor type two (TNF-RII) gene have been identified, but their biological consequences remain poorly understood. The aim of this study was to determine whether polymorphism in the analysed loci of the bovine TNF-RII gene is linked with the size of cell subpopulations naturally infected with bovine leukaemia virus (BLV) which serve important immune functions in the host. Material and Methods: Samples originated from 78 cows. Polymorphisms in the studied gene were determined by PCR-RFLP and DNA sequencing by capillary electrophoresis. BLV infection was diagnosed by the immunofluorescence (IMF) technique and nested PCR. Cell subpopulations were immunophenotyped with IMF. Results: Similar and non-significant differences in the average percentages of TNFα+, IgM+TNFα+, and CD11b+TNFα+ cells infected with BLV were noted in individuals with various genotypes in the polymorphic sites g.-1646T > G and g.16534T > C of the TNF-RII gene, and significant differences in the percentages of these subpopulations were observed between selected microsatellite genotypes (g.16512CA(n)). Conclusion: STR polymorphism and the number of CA dinucleotide repeats in intron 1 of the TNF-RII gene influence the frequency of TNF+, CD11b+TNF+, and IgM+TNF+ subpopulations naturally infected with BLV. Polymorphism in the gene’s other two sites do not affect the size of these cell subpopulations.

Introduction: The study sought to characterise antimicrobial resistance among coagulase-negative Staphylococcus (CNS) species recovered from broiler chickens and turkeys in Poland including the presence of 12 antimicrobial resistance genes and five classical genes of staphylococcal enterotoxins. Material and Methods: A panel of 11 antimicrobial disks evaluated the phenotypic sensitivity of the tested strains to antibiotics. Five multiplex PCR assays were performed using primer pairs for specific detection of antibiotic resistance genes and staphylococcal enterotoxin A to E genes. Results: Selected antimicrobial agent susceptibility testing revealed 100% of such in in vitro conditions to cefoxitin among strains of Staphylococcus sciuri and S. chromogenes. The blaZ (for ß-lactam) and mecA (for methicillin resistance) genes were in 58.3% and 27.5% of strains, respectively. Among genes resistant to tetracyclines, tetK was most frequent. Fewer (CNS) strains showed genes resistant to macrolides, lincosamides, and florfenicol/chloramphenicol. Multiplex PCR for classical enterotoxins (A-E) detected the see gene in two S. hominis strains, while the seb gene producing enterotoxin B was found in one strain of S. epidermidis. Conclusion: CNS strains of Staphylococcus isolated from poultry were either phenotypically or genotypically multidrug resistant. Testing for the presence of the five classical enterotoxin genes showed that CNS strains, as in the case of S. aureus strains, can be a source of food intoxications.

Introduction: Pathomorphological changes in the lungs, stomach, and small intestines of wild boars infected with Metastrongylus spp., Ascarops strongylina, and Macracanthorhynchus hirudinaceus were investigated. Material and Methods: Dissection of 11 wild boars was performed, and parasitised organs were histologically investigated by common techniques. Results: Macroscopic lesions in the lungs infected with Metastrongyus spp. were seen within the apical parts of the large lobes, irregular in form, pale greyish in colour, and compact in consistency. The main pathohistological findings were: the presence of parasite forms, and lymphocytes and neutrophils in the lumen of bronchi and bronchioles, desquamation of the bronchial and bronchiolar epithelium, emphysema, thickening of alveolar septa, hyperaemia, alveolitis, infiltration of the interstitial tissue with giant cell, monocytes and eosinophils, and peribronchial and disseminated lymphoid hyperplasia. The principal observations accompanying infection with A. strongylina were inflammation and focal mucosal damage in the stomach, the latter clearly demarcated from the surrounding tissues. Severe injuries in the place of attachment of M. hirudinaceus to the wall of the small intestine were seen. Intestinal villi, underlying mucosa, and submucosa were destroyed, and an intense inflammatory reaction was present. Conclusion: The histopathological lesions showed wide diversity, varying from mild to severe; but none of them were lethal.

Introduction: Farm mink (Neovison vison) can be naturally exposed to T. canis and T. leonina pathogens on the farm. If mink were hosts, it would imply some veterinary public health as well as animal welfare issues. For this reason, the aim of the study was to determine whether mink might be definitive or paratenic hosts of these parasites. Material and Methods: Four groups of mink were infected with both parasite species using larvated eggs or feed containing mouse tissue previously infected with the parasites. Following inoculation, the infections were monitored in vivo by faecal examination for 14 weeks p.i., and then western blotting and ELISA were performed. Results: Coprology did not reveal any canine roundworm eggs, neither were nematodes found in mink intestines during post mortem examination. The specific IgG antibodies recognising excretory/secretory (ES) antigens of both parasite species were identified in mink sera. Single T. leonina tissue larvae were found in digested organs. Conclusions: Our results confirm that farm mink may contribute both T. canis and T. leonina infections. It was proved that farm mink were not their definitive hosts, and therefore mink faeces need not be considered a source of canine roundworm eggs in any soil it fertilises. Nonetheless, as farm mink may be a paratenic host for both parasite species, this may have some impact on the health and welfare of infected animals.

Introduction: Canine roundworm T. canis and T. leonina infections were investigated in experimentally infected farm mink (Neovison vison) to describe the pattern of pathological lesions in this paratenic host. Material and Methods: Infections in mink developed following ingestion of embryonated eggs of either parasite or mice tissue infected with both parasite species. Results: Comparative analysis of haematoxylin- and eosin-stained slides showed essential differences among the experimental groups. The lesions observed included eosinophil and mononuclear inflammatory infiltrates of the intestinal wall and local lymph nodes, inflammation and haemorrhages in liver tissues, and interstitial inflammation and mineralisation of the kidneys and lungs. Larvae migrating through the minks’ bodies also caused particularly salient enlargement of lymphoid follicles in the spleen and inflammatory infiltrates of mononuclear cells in skeletal and heart muscles. Conclusions: It is assumed that histopathological lesions appeared as a local and general host response to invasive L3 T. canis and T. leonina larvae migrating through the tissues of infected farm mink. Interestingly, mink infected with embryonated eggs had more pronounced lesions than animals infected with tissue larvae. Detailed histopathological examinations of parenchymal organs and striated muscles revealed lesions resembling those observed in other paratenic host species due to toxocarosis.

The cyclooxygenase-2 (COX-2) enzyme catalyses the first stage of biosynthesis of prostanoids, proteins that are implicated in various physiological and pathological processes in humans and animals. The expression of COX-2 increases significantly during pathological processes accompanied by inflammation, pain and fever. Overexpression of COX-2 was determined in tumour tissues, which suggests that this enzyme participates in oncogenesis. In this paper the topics discussed are mechanisms regulating COX-2 expression, COX isoforms, their role in the body and the oncogenic mechanisms triggered by the overexpression of COX-2, including inhibition of apoptosis, intensification of neoangiogenesis, increased metastatic capacity, and weakening of the immune system. The significance of and the mechanisms by which COX-2 participates in oncogenesis have been studied intensively in recent years. The results are highly promising, and they expand our understanding of the complex processes and changes at the molecular, cellular and tissue level that promote oncogenesis and cancer progression. Notwithstanding the knowledge already gleaned, many processes and mechanisms have not yet been elucidated in human medicine and, in particular, in veterinary medicine. Further research is required to develop effective tumour diagnostic methods and treatment procedures for humans and animals.

Introduction: Canine transmissible venereal tumour (CTVT) is a sexually transmitted tumour affecting dogs worldwide, imposing a financial burden on dog owners. A stable culture cell line in continuous passages for >18 months has only been achieved once. The present study investigated a stable CTVT cell line isolated from a bitch and its potential as a vaccine. Material and Methods: A biopsy from a 2-year-old mongrel bitch with CTVT was obtained for histopathological confirmation and isolation of tumour cells. The isolated cells were cultured to passage 55 and characterised by flow cytometry, with karyotyping by GTG-banding and by PCR detection of myc S-2 and LINE AS1. The isolated CTVT cell line was also used as a preventive vaccine in a canine model. Results: Histopathological analysis of the isolated tumour cells revealed typical CTVT characteristics. Constant proliferation and stable morphological characteristics were observed during culture. Phenotypic analysis determined the expression of HLA-DR⁺, CD5.1⁺, CD14⁺, CD45⁺, CD83⁺, CD163⁺, and Ly-6G-Ly-6C⁺. GTG-banding revealed a mean of 57 chromosomes in the karyotype with several complex chromosomal rearrangements. LINE-c-myc insertion in the isolated CTVT cell line at 550 bp was not detected. However, a 340-bp band was amplified. Isolated CTVT cell line inoculation at a concentration of 1×10⁸ did not induce tumour growth in bitches, nor did a challenge with primary CTVT cells. Conclusion: The present study successfully identified and isolated a stable CTVT cell line that may be useful in CTVT prevention.

Introduction: Antimicrobial peptides (AMP) are a large group of innate immune effectors, which apart from antimicrobial activity show immunomodulative properties. Platelet-rich plasma (PRP) is a source of autologous growth factors and is used for stimulation of bone and soft tissue healing. The purpose of this study was to assess the influence of PRP and AMP extract on ovine monocyte-derived macrophage cultures. Material and Methods: The study was conducted on ovine macrophages (Mfs) previously stimulated with LPS or dexamethasone and then with preparations of PRP or AMP. Following activation of the Mfs their morphological and functional features were assessed. Results: The study revealed pro-inflammatory influence of both examined preparations on Mfs cultures on the basis of morphology, ROS generation and arginase activity. Both preparations enhanced the pro-inflammatory response of cultured Mfs. Conclusion: This activity may intensify the antimicrobial action of Mfs, however, in cases of excessive and prolonged inflammation the use of these preparations should be limited.

Introduction: The clinical symptoms of portosystemic shunts (PSSs) and hepatic microvascular dysplasia (HMD) – portal vein hypoplasia (PVH) in dogs are similar. PSSs are abnormal vascular connections between the portal vein system and systemic veins. HMD is a very rare developmental vascular anomaly, recognisable during histopathological examination. The study aim was to assess the prevalence of HMD–PVH and hepatocellular and vascular pathologies in the liver. Material and Methods: Liver biopsies from 140 dogs (of different breeds and both sexes) arousing clinical suspicion of PSS were examined histopathologically. Results: An initial PSS diagnosis was confirmed in 125 dogs (89.29%). HMD–PVH was found in 12.32% of dogs, as an isolated disease in 9.29%, especially in Yorkshire terriers, and with extrahepatic PSS in 6.67%. Histopathological analysis of muscles around sublobular veins showed that HMD cases presented hypertrophy or hypertrophy with fibrosis. In 2.17% of all dogs with liver vascular developmental disorders calcification was visible around vessels (without correlation by degenerative changes in those vessels), suggesting prior onset of deep metabolic disorders. Clinical suspicion of PSS was also formed upon quite different pathological processes in young dogs. Conclusion: Histopathological findings diagnosed the type of vascular anomalies (PSS or HMD–PVH) or other pathological changes conclusively, therefore detailed hepatic histopathology is an indispensable component of the clinical diagnostic process.

Introduction: The aim of this research was to provide a detailed description of the morphology, topography, and histometry of rabbit accessory genital glands. Material and Methods: Seven male New Zealand White rabbits, 3–4 months of age and weighing 2.1–3 kg were used for the study. The whole urethra from the urinary bladder to the external urethral orifice accompanied by accessory genital glands was sliced at intervals of 1 mm. The serial sections were prepared with haematoxylin-eosin (H&E) and Movat–Russell modified pentachrome stain. Results: A detailed description of the morphology and morphometry was provided. The topography of the organs was explained on the basis of characteristic cross-sections on histological slides. The inconsistent nomenclature and descriptions of these glands by different authors were also discussed. Conclusion: The morphometric analysis indicated that some of the glands described have similar dimensions in different individuals, while others like paraprostates revealed high diversity in the number of lobes, their size, and their structure. The accessory glands are also good topographic markers which precisely define the segment of the urethra. The terms “proprostate”, “prostate”, and “paraprostates” as the nomenclature of the prostate complex reflect the location of these glands well and indicate their common origin and function.

Introduction: The aim of the study was to assess the usefulness of duplex Doppler to objectify swine renal arterial flow in physiological conditions. The pig kidney was selected for its morphological similarities to the human and for the results therefore offering data to wider research. Material and Methods: Six White Large x Landrace sows, of 48.5–53 kg b.w. were used. Vascular flow parameters were acquired with a convex probe USG device with a duplex Doppler function using pulsed waves (frequency range of 5–7.5 MHz). Segmental kidney arterial flow was measured. Results: The RI values were within the 0.57 (min) to 0.6 (max) range, ̄x_RI was 0.58 (±0.014), and the SD² value was 0.0002. The PI index values ranged from 1.21 (min) to 1.3 (max), and ̄x_RI was 1.24 (±0.035). The value of SD² was 0.00123. In the S/D index, the results fell between 2.2 (min) and 2.49 (max), with ̄x_S/D of 2.29 (±0.117). The value of variance SD² was 0.0139. A double analysis of correlation between indices showed this to be almost certain and highly positive as confirmed by high correlation coefficients: r RI & PI 0.857, rho_{RI & PI} 0.739, r_{RI & S/D} 0.930, rho_{RI & S/D} 0.941, r_{PI & S/D} 0.913, and rho_{PI & S/D} 0.754. The segmental kidney arterial flow spectrum evinced falls in PSV and PSV/LDV ratio, also noticed in the filling of the spectral window, comparing the renal to an interlobar artery. Conclusion: Swine were selected because of renal anatomical and haemodynamic similarity to humans. The most relevant values and indices approximated those in humans. The study anaesthetic protocol had a minor influence on the average RI, PI, and S/D indices.

Introduction: The thyroid and parathyroid glands play a major role in maintaining physiological homeostasis in all vertebrates. Reptiles have plasma concentrations of thyroid hormones far lower than mammals. Low levels of these hormones in reptiles impede thyroid hormone detection with assays designed for the higher levels of mammals. The aim of this study was to explore teaming this with ultrasound imaging of the thyroid to appraise glandular function. Material and Methods: Thyroid function of four pond sliders was evaluated based on the results of T4 analyses and ultrasound. Results: The concentrations of T4 varied considerably between the examined animals from <9 nmol/L to >167.3 nmol/L. Ultrasound examination revealed uniform echogenicity and a smooth outline of the thyroid gland in all animals. Conclusion: Monitoring of thyroid function based on T4 and electrolyte concentrations is helpful in assessing the health and living conditions of reptiles, which is important in veterinary practice but problematic. Ultrasound examinations are useful in diagnosing changes in gland structure, such as tumours and goitres, and a combination of both methods supports comprehensive assessments of the anatomy and function of the thyroid gland.

Introduction: Clinical doses of anaesthetic agents were administered to rabbits and effects on the brain, heart, and liver were investigated biochemically and histopathologically. Material and Methods: The rabbits were randomly divided into three main groups (16 rabbits each) and each group into study (n = 8) and control (n = 8) groups. All study group rabbits received 3 mg/kg of midazolam (M) intramuscularly. Group 1.1 (M) received nothing further, group 2.1 (MK) also received 25 mg/kg of ketamine, and group 3.1 (MKI) besides ketamine was also given 2% isoflurane to induce anaesthesia for 30 min. NaCl solution in the same volume as midazolam and ketamine was injected into the controls. Results: In clinical evaluation significant differences were detected in respiratory and heart rates. In blood gas analysis the PO2 and PCO2 values showed statistical differences in anaesthesia intervals. Significant biochemical value changes were recorded in creatine kinase-Mb, glucose, and total protein. Histopathological liver examinations revealed higher total apoptotic and normal cell numbers in the MK than in the M and MKI groups. Apoptotic cell numbers were statistically significant in M and MK groups. Conclusion: Anaesthetic agents may increase programmed apoptosis. The MKI anaesthetics combination was found to cause less cell destruction in general than the other study groups. It was indicated that MKI was the safer anaesthetic combination in rabbits.

Introduction: Tiletamine-xylazine-tramadol (XFM) has few side effects and can provide good sedation and analgesia. Adenosine 5’-monophosphate-activated protein kinase (AMPK) can attenuate trigeminal neuralgia. The study aimed to investigate the effects of XFM and its specific antagonist on AMPK in different regions of the brain. Material and Methods: A model of XFM in the rat was established. A total of 72 Sprague Dawley (SD) rats were randomly divided into three equally sized groups: XFM anaesthesia (M group), antagonist (W group), and XFM with antagonist interactive groups (MW group). Eighteen SD rats were in the control group and were injected intraperitoneally with saline (C group). The rats were sacrificed and the cerebral cortex, cerebellum, hippocampus, thalamus, and brain stem were immediately separated, in order to detect AMPKα mRNA expression by quantitative PCR. Results: XFM was able to increase the mRNA expression of AMPKα1 and AMPKα2 in all brain regions, and the antagonist caused the opposite effect, although the effects of XFM could not be completely reversed in some areas. Conclusion: XFM can influence the expression of AMPK in the central nervous system of the rat, which can provide a reference for the future development of anaesthetics for animals.

Introduction: Significant improvement of sperm motility within one month effected by oral supplementation of selenium and vitamin E was described in four infertile male dogs which failed to conceive in their last three matings with different bitches. Material and Methods: The dogs (a Golden Retriever, an English Cocker Spaniel, and two Tibetan Mastiffs) were supplemented daily with selenium (Se) (0.6 mg/kg organic Se yeast) and vitamin E (vit. E) (5 mg/kg) per os for 60 days. Semen was collected on days 0, 30, 60, and 90. The sperm concentration and motility parameters were evaluated by the CASA system, sperm morphology was explored by Diff-Quick staining, and live and dead spermatozoa were differentiated by eosin/nigrosin staining. The concentrations of Se and vit. E were measured in peripheral blood serum on semen collection days. Results: Before administration, the concentrations of Se in blood plasma were low (86.0–165.0 µg/L). After 30 days of treatment there was an observable improvement in total and progressive sperm motility and kinematic parameters (VAP, VSK, VCL, ALH, BCF, and RAPID). The percentages of live and normal morphology sperm cells were also higher. There was also an observable increase in Se and vitamin E concentrations in blood serum. Bitches were successfully mated and delivered four to six puppies. Conclusion: Supplementation with Se and vit. E improved rapid sperm motility and restored fertility in infertile dogs with low Se status.

Introduction: Breeding profiles at the periparturient stage in red foxes which mated naturally or were subjected to artificial insemination were retrospectively surveyed using 130 vixens during their reproductive seasons of 2012–2017 in Japan. Material and Methods: Natural mating vixens were encouraged a maximum of three times with the same male, while artificial insemination was conducted using frozen-thawed semen with the bovine semen extender as a diluent. Results: With natural mating, conception rates after one, two, and three copulations were 55.8%, 68.0%, and 85.7%, respectively, showing a significant difference between the rates for one and three copulations. Conception rates with artificial insemination were 82.4%. Mean gestation periods were between 52.1 and 53.3 days in all groups. Mean litter sizes were 3.7–4.3 cubs with natural mating, and 4.4 cubs with artificial insemination. Although some sporadic and inconsistent changes in litter sizes were noted between primiparous and multiparous groups, these were of doubtful clinical importance. Conclusion: This is the first report from Japan concerning basic breeding events of red fox vixens in captivity.