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

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.).

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: 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.

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.

Introduction: Comparative analysis of the karyotype structure was made in two hedgehog species: the northern white-breasted hedgehog (Erinaceus roumanicus) and the African pygmy hedgehog (Atelerix albiventris). Material and Methods: The cytogenetic analysis used differential staining techniques (DAPI, Ag-NOR, and C-banding/DAPI) and sequential QFQ/FISH banding with NOR20 and TEL20 probes which showed 45S rDNA and (TTAGGG)n repeat sequences, respectively, on hedgehog chromosomes. Results: It was confirmed that the somatic cells of the hedgehogs have a constant number of chromosomes (2n = 48,XY). Differences were observed in the NOR number between the species. NORs were identified on three autosome pairs in the northern white-breasted hedgehog and on only two pairs in the African pygmy hedgehog. Chromosome analysis by C-banding/DAPI showed large segments of heterochromatin rich in A-T pairs on three autosome pairs in both the northern white-breasted and African pygmy hedgehogs. The heterochromatin segments encompassed large fragments of the longer arm of chromosome pairs 13, 14 and 20. The (TTAGGG)n repeat sequences on the hedgehog chromosomes were only observed in the terminal position of all the chromosomes in both species. Conclusion: Our observations provide new information on the level of diversity within the Erinaceidae family. | hedgehog; chromosomes; heterochromatin; nucleolar organisers; telomere | 140 | 353-358

Comparative analysis of the karyotype structure was made in two hedgehog species: the northern white-breasted hedgehog (Erinaceus roumanicus) and the African pygmy hedgehog (Atelerix albiventris). The cytogenetic analysis used differential staining techniques (DAPI, Ag-NOR, and C-banding/DAPI) and sequential QFQ/FISH banding with NOR20 and TEL20 probes which showed 45S rDNA and (TTAGGG)ₙ repeat sequences, respectively, on hedgehog chromosomes. It was confirmed that the somatic cells of the hedgehogs have a constant number of chromosomes (2n = 48,XY). Differences were observed in the NOR number between the species. NORs were identified on three autosome pairs in the northern white-breasted hedgehog and on only two pairs in the African pygmy hedgehog. Chromosome analysis by C-banding/DAPI showed large segments of heterochromatin rich in A-T pairs on three autosome pairs in both the northern white-breasted and African pygmy hedgehogs. The heterochromatin segments encompassed large fragments of the longer arm of chromosome pairs 13, 14 and 20. The (TTAGGG)ₙ repeat sequences on the hedgehog chromosomes were only observed in the terminal position of all the chromosomes in both species. Our observations provide new information on the level of diversity within the Erinaceidae family.

Comparative analysis of the karyotype structure was made in two hedgehog species: the northern white-breasted hedgehog (Erinaceus roumanicus) and the African pygmy hedgehog (Atelerix albiventris).

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

Gastrointestinal nematodes pose a threat to animal health and affect farmers by negatively impacting farm management.

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

Strains of Leptospira interrogans belonging to two very closely related serovars, Icterohaemorrhagiae and Copenhageni, have been associated with disease in mammalian species and are the most frequently reported agents of human leptospirosis. They are considered the most pathogenic serovars and represent more than half of the leptospires encountered in severe human infections.

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

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, ̄xRI 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 ̄xRI 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 ̄xS/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, rhoRI & PI 0.739, rRI & S/D 0.930, rhoRI & S/D 0.941, rPI & S/D 0.913, and rhoPI & 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: Grazing cattle are vulnerable to the harmful effects of gastrointestinal parasites. Organically farmed cattle are even more so because conventional antiparasitic treatments are restricted, yet parasite infection patterns in Polish organic herds remain poorly documented. Material and Methods: Imported beef cattle were studied during the pasture season in four organic herds in southern Poland. The McMaster quantitative flotation method was used to estimate infection prevalence (P, %) coproscopically and to quantify intensities of coccidia oocyst output (Ic, OPG) and nematode egg output (In, EPG) per gram of faeces. The qualitative sedimentation method was applied to assess the presence of digenean eggs. Coccidial species of the Eimeria genus were identified by sporulation, and nematodes of the Strongylida order by larvoscopy. Digenean Paramphistomatidae were identified by morphological examination of adult fluke specimens obtained at slaughter from a sick heifer in one of the studied herds and by molecular analysis of the flukes’ internal transcribed spacer 2 ribosomal DNA. Results: The prevalence of Eimeria infection was P = 28.9 (23.8–34.5)%, and the mean Ic was 287 (113–793) OPG. Calves were most heavily infected, mainly with E. bovis and E. zuernii. The prevalence of nematode infections reached P = 46.0 (40.2–51.5)%, and the mean In was 113 (88–147) EPG. Haemonchus placei dominated over Ostertagia sp. and Trichostrongylus axei, and the most infected were first-time grazing yearlings. Paramphistome eggs were confirmed in only one herd. Morphological and PCR analysis of the adult rumen flukes revealed the presence of Calicophoron daubneyi (Dinnik, 1962) in this herd. Conclusion: This is the first Polish evidence of C. daubneyi, and it heralds an enhanced surveillance need regarding this highly pathogenic digenean. | helminths, paramphistomes, Strongylida, coccidia, beef cattle | 140 | 1-11 | 69

Grazing cattle are vulnerable to the harmful effects of gastrointestinal parasites. Organically farmed cattle are even more so because conventional antiparasitic treatments are restricted, yet parasite infection patterns in Polish organic herds remain poorly documented.

Introduction: Diarrhoea in growing-finishing pigs is a common problem of commercial pig farms. Among many causative factors, porcine circovirus type 2 (PCV2) is one considered an important pathogen in modern pig production. The aim of the study was to verify if PCV2 was responsible for antibiotic non-responsive diarrhoea and wasting in pigs. Material and Methods: A total of 13 dead pigs aged between 12 and 15 weeks from three Polish farms with persistent herd symptoms suggestive of PCV2 infection were provided for evaluation. Sections of lymph nodes and intestines were analysed by in situ hybridization (ISH) for PCV2 and histopathological examination. Faeces and intestinal scrapings were tested for Lawsonia intracellularis and Brachyspira hyodysenteriae by real-time PCR and for parasitic infection by flotation and decantation. Results: ISH and histopathological examination showed that all pigs were PCV2 systemic disease negative. Swine dysentery was confirmed by real-time PCR on two farms, and proliferative enteropathy on one farm. In histological examinations, erosions of the caecal and colonic mucosa were found, together with cysts and trophozoites of Balantidium coli. The protozoa were present in the intestinal lumen and mucosa. B. coli cysts were identified in faeces from all examined pigs. Conclusion: These results suggest that monitoring of B. coli infections should be an additional measure of control and prevention of gastrointestinal tract disorders in modern swine husbandry. | pigs, diarrhoea, PCV2, Balantidium coli, farm water quality | 140 | 1-6 | 65

Diarrhoea in growing-finishing pigs is a common problem of commercial pig farms. Among many causative factors, porcine circovirus type 2 (PCV2) is one considered an important pathogen in modern pig production. The aim of the study was to verify if PCV2 was responsible for antibiotic non-responsive diarrhoea and wasting in pigs. A total of 13 dead pigs aged between 12 and 15 weeks from three Polish farms with persistent herd symptoms suggestive of PCV2 infection were provided for evaluation. Sections of lymph nodes and intestines were analysed by in situ hybridization (ISH) for PCV2 and histopathological examination. Faeces and intestinal scrapings were tested for Lawsonia intracellularis and Brachyspira hyodysenteriae by real-time PCR and for parasitic infection by flotation and decantation. ISH and histopathological examination showed that all pigs were PCV2 systemic disease negative. Swine dysentery was confirmed by real-time PCR on two farms, and proliferative enteropathy on one farm. In histological examinations, erosions of the caecal and colonic mucosa were found, together with cysts and trophozoites of Balantidium coli. The protozoa were present in the intestinal lumen and mucosa. B. coli cysts were identified in faeces from all examined pigs. These results suggest that monitoring of B. coli infections should be an additional measure of control and prevention of gastrointestinal tract disorders in modern swine husbandry.

Diarrhoea in growing-finishing pigs is a common problem of commercial pig farms. Among many causative factors, porcine circovirus type 2 (PCV2) is one considered an important pathogen in modern pig production. The aim of the study was to verify if PCV2 was responsible for antibiotic non-responsive diarrhoea and wasting in pigs.