Introduction: Pregnancy is a physiological state in which the immune system undergoes certain changes. On the one hand, by depleting cell defence mechanisms, it favours development and maintenance of the pregnancy. At the same time cells of the immune system ensure resistance to many risk factors, including infectious agents. Material and Methods: The study was carried out on 24 Polish Konik breed mares which were divided into two equal groups. The first group (group I) included mares living in the reserve. The second group (group II) comprised mares maintained under conventional conditions in the stables. The blood samples were collected for the first time in the perinatal period, i.e. 2 weeks before parturition (trial 0), then within the first 24 h after delivery, and then on 7th and 21st day after foaling. Flow cytometric analysis of lymphocyte expressing TCD4+, TCD8+, CD2+, and MHC class II antigens was performed. Results: Before the delivery, in group I there was a significantly higher CD4:CD8 ratio compared to group II (P ≤ 0.05). Similarly, significantly increased CD4:CD8 ratio in group I was noted within 24 h after parturition (P ≤ 0.001) and it was also observed on 7th day (P ≤ 0.03) and 21st day after foaling (P ≤ 0.02). In the first 24 h after parturition, a significant decline of lymphocytes CD8+ (P ≤ 0.02) was noted. No significant differences in terms of lymphocytes CD2+ and CD3+ were observed. Expression of MHC-II molecules before and after the parturition was higher in group I compared to group II; however, the difference between the groups was not significant. Conclusion: The results obtained indicate that mares living in the reserve display higher activity of cell defence mechanisms. | mares, pregnancy, immunity, lymphocytes, major histocompatibility complex | 20 | 217-221 | 2

Introduction: Strains of Leptospira interrogans belonging to two very closely related serovars, Icterohaemorrhagiae andCopenhageni, have been associated with disease in mammalian species and are the most frequently reported agents of humanleptospirosis. They are considered the most pathogenic serovars and represent more than half of the leptospires encountered insevere human infections. Material and Methods: Nineteen such isolates from the United Kingdom – human, domestic andwildlife species – were typed using three monoclonal antibodies (F12 C3, F70 C14 and F70 C24) in an attempt to elucidate theirepidemiology. They were further examined by restriction endonuclease analysis (REA), multiple-locus variable-number tandemrepeat analysis (MLVA) and lic12008 gene sequence analysis. Results: Monoclonal antibody F12 C3, which is highly specificfor Icterohaemorrhagiae and Copenhageni, confirmed that all the strains belonged to these two serovars. Sixteen strains wereidentified as Copenhageni and three as Icterohaemorrhagiae serovar. Only one restriction pattern type was identified, thusconfirming that REA is not able to discriminate between the Icterohaemorrhagiae and Copenhageni serovars. Variable-numbertandem-repeat analysis found three loci with differences in the repeat number, indicating genetic diversity between Britishisolates. Sequences of the lic12008 gene showed that all isolates identified as the Icterohaemorrhagiae serotype have a singlebase insertion, in contrast to the same sequences of the Copenhageni serotype. Conclusion: Copenhageni is the predominantserovar in the Icterohaemorrhagiae serogroup isolated in British Isles. There is a genetic diversity of MLVA patterns of theisolates but no genetic tool used in the study was able to determine serovars. | Leptospira, leptospirosis, Icterohaemorrhagiae, Copenhageni, animal pathogens | 200 | 529-536 | 4

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 PI 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. | swine, kidney, resistance index, pulsatility index, systolic/diastolic index | 140 | 259-265 | 2

The aim of this study was to identify mutations in the D-loop region of mtDNA in head, neck, and limb tumours in dogs,and determination of their relationship with the process of neoplastic transformation. Blood and tumour tissue samples from 19dogs with diagnosed sporadic malignant tumours were analysed. DNA extraction, amplification, and sequencing of the mtDNAD-loop, and bioinformatic analyses were performed. Five mutations and 19 polymorphisms were observed in 68.42% of alltumours. Polymorphic variants were noted in 42.86% of the head and neck tumours and in 58.33% of the limb tumours.Mutations were observed in 21.05% of dogs. The mutations were found in 28.57% of the head and neck tumours and in 16.66%of the limb tumours. The mutations were identified in 50% of the studied epithelial cancers. In the mesenchymal tumours, nomutations in the D-loop region were observed. Mitochondrial haplotype A17 was found in over 40% cases of limb tumours. Noassociation between the age, breed, sex, type of tumour, and detected polymorphic variants were observed. Different mutationalchanges in the D-loop sequences of mtDNA identified in the blood and tumour tissues may indicate a relationship between thetype of tumour and individual changes in the D-loop nucleotide sequences of mtDNA. | dog, tumour, D-loop, mtDNA, mutations | 20 | 631-637 | 4

Introduction: Cadmium and zinc are often found in aquatic environment and may accumulate in living organisms. The aim of this study was to evaluate the genotoxic effect of Cd, Zn, and their binary mixture on the peripheral blood erythrocytes of Prussian carp (Carassius gibelio B.). Material and Methods: The fish were exposed to 4.0 mg/L Cd, 4.0 mg/L Zn or a mixture of 4.0 mg/L Cd and 4.0 mg/L Zn for a period of 14, 21 or 28 days. Genotoxic effects were investigated in peripheral blood cells using the comet assay and the erythrocyte micronucleus assay. Results: The results demonstrated that the frequencies of micronuclei (MN) and both nuclear and cellular abnormalities in erythrocytes were significantly higher in all exposure groups as compared to the control group. The fish exposed to the mixture of Cd and Zn presented the highest frequency of MN. Furthermore, there was a decrease in the frequency of MN and an increase in the occurrence of DNA integrity defects (DNA damage) with longer time of exposure to the metals studied. Conclusion: Erythrocyte micronucleus and comet assays confirmed the genotoxicity of Cd and Zn. The results of the tests applied (which showed considerable variability) suggest the involvement of various toxicity mechanisms. Therefore, an integrative and comprehensive approach, using a set of assays for toxicity profile determination, should be adopted during ecotoxicological studies and environmental risk assessment pertaining to these elements. | genotoxicity, heavy metals, erythrocyte micronucleus assay, comet assay, Prussian carp. | 140 | 619-628 | 4

Introduction: Gastrointestinal nematodes pose a threat to animal health and affect farmers by negatively impacting farm management. Material and Methods: The study was conducted on a sheep farm with suspected reductions in the efficacies of anthelmintics. Efficacy was determined using in vivo faecal egg count reduction, in vitro egg hatch (EHT) and larval development (LDT) tests. In the first phase, 60 sheep were equally split into six groups. Group 1 received the recommended dose of albendazole (ALB), group 2 received the same after fasting for 24 h, group 3 received the dose divided into two halves at 6 h intervals, group 4 received a double dose of ALB, and group 5 received the recommended dose of ivermectin (IVM). Group 6 served as a control. The second phase of the experiment had two groups: one treated with levamisole (LEV) and a control group. Faecal samples were collected from all sheep. Results: No reduction of egg output was observed in the groups treated with single, double, or divided doses of ALB, but one of 13.7–16.9% was noted in the fasting group. Efficacy in the IVM group ranged from 31.50 to 39.97%. The mean concentrations sufficient to prevent 50% of the eggs from hatching in the in vitro EHT and the mean concentrations in which the development of larvae to the L3 stage was inhibited by 50% in the LDT exceeded established thresholds for benzimidazoles and IVM. Haemonchus contortus was the only species identified after treatment. The LDT did not indicate the presence of resistance to LEV. All animals treated with LEV were negative for eggs 10 d after treatment. Conclusion: Resistance to ALB and IVM in Haemonchus contortus was confirmed. Alternative approaches to improve the efficacies of benzimidazole did not sufficiently increase the efficacy, but LEV was an efficient anthelmintic treatment. | sheep, drug resistance, alternative treatment, Haemonchus contortus, levamisole | 200 | 575-581 | 4

Background: The equine colic, which is caused by the presence of enteroliths that are most often found in the small or large colon, is typical for certain geographical regions (dry and hot climate). A diet rich in alfalfa is one of the highest risk factors. The earliest symptoms include weight loss and repeated episodes of colic pain. Aims: To present the results of operative treatment of 15 horses with enteroliths in Saudi Arabia. Methods: Fifteen purebred Arabian horses in Saudi Arabia, aged between 2 and 18 years, were treated. Decision about the surgery was based on clinical exam, ultrasound and rectal examination. The surgery was done on recumbent position in every case, under general inhalation anesthesia performed with izofluran. Results: Midline laparotomy was performed in all cases. Additional left flank laparotomy was performed in one horse, in which the stone was located in the proximal part of the small colon and parainguinal laparotomy was performed in 1 horse, in which the stone was located distally in the small colon. In each case, pelvic flexure enterotomy was performed in order to empty the large colon. Additionally, four horses underwent ventral colon enterotomy due to the presence of large stones. Small colon enterotomy was performed in 9 horses. In 12 cases treatment outcome was good and in 3 - poor. Chemical analysis of the stones showed similar results: calcium, calcium oxalate, ammonium, phosphates, and magnesium (Mg) were obtained in all these cases but there were quantitative differences accounting for 15 to 30%, 10 to 20%, 10%, 20 to 40%, and 10 to 15%, respectively. Conclusion: The results of surgery are generally good if stones are located in the large colon, but the prognosis is worse if they are located in the small colon, particularly in its proximal part. There is a huge importance of X-ray examination, which allows accurate diagnosis for locating the enteroliths and making a decision about surgery. | Enterolithiasis, Equine colic, Laparotomy | 20 | 270-276 | 4

This study aimed to determine whether the level of fearfulness and cortisol reactivity influenced spatial learning performance in primitive mountain Hucul horses. The Huculs had to overcome particularly difficult, sometimes dangerous conditions to survive over centuries. We hypothesized that they were familiar with danger and should learn easily. The study involved 22 mares. They were divided into groups of different fearfulness levels according to behavioral score and cortisol concentration increase in the saliva, both of which were determined in a test of a novel object. In the next step, the mares were subjected to a spatial learning test in a maze with food reward at the end, conducted in 2 trials, on consecutive days. Their spatial learning performance was assessed with an error score and time taken to complete the maze. The study found that both the level of fearfulness and cortisol concentration increase varied in the mares. The spatial learning performance of mares that had low, medium and high levels of fearfulness and cortisol increase was similar within a trial, although animals displaying a medium fearfulness level or moderate cortisol change achieved better results in the second trial than in the first trial. These results indicate a weak influence of the fearfulness and cortisol reactivity on spatial learning performance in Huculs which may suggest an effect of the origin and historical environment of the breed. | Primitive horse, Behavior, Fearfulness, Spatial learning, Maze test, Cortisol | 100 | 10-17

A trial was performed in a swine research facility to ascertain the protection provided by a polyvalent Actinobacillus pleuropneumoniae (APP) bacterin containing serotypes 1, 3, 5 and 9. The test animals consisted of 60, eight-week-old, piglets, which were randomly divided into four main groups. The four main groups were further divided into three sub-groups (I, II, III) of five pigs each. Subgroup I was vaccinated intramuscularly, sub-group II was vaccinated subcutaneously, and sub-group III served as the unvaccinated control group. Each main group was challenged with a single APP serotype (1, 3, 5 or 9). Criteria for evaluation of the bacterin efficacy were mortality, lung lesions, pleural adhesions, and isolation of APP from tonsil or lung. Significant effects of vaccination over nonvaccination were reduced mortality, lung lesions, pleural adhesions, and isolations of APP from tonsil and lung. There were no significant differences between the intramuscular and subcutaneous routes of vaccination. It was concluded that the four-way APP bacterin used in this study provided satisfactory protection against homologous challenge. Evidence of protection was lower mortality and lung lesions and increased daily weight gains in vaccinates as compared with controls. | 16 | 233–238 | 35 (4)