Volumen 59 (2015): Heft 2 (June 2015)

Birds are one of the most interesting and most colourful groups of animals, but they can also be a source of zoonotic factors dangerous for humans. This paper describes the threats to human health from contact with birds. The most vulnerable occupational groups associated with birds are veterinarians, owners of poultry farms, breeders of ornamental birds, zoo personnel, and poultry slaughterhouse workers. Ornithosis is the most dangerous zoonosis of the avian bacterial diseases. Among other hazardous bacterial factors, Salmonella and Campylobacter are responsible for gastrointestinal diseases. Avian influenza is the most dangerous of the viral diseases. It should be noted, however, that avian influenza is a disease of birds, not humans. The recent threat which has appeared is infection with West Nile virus. The results of serological examinations of birds and humans indicate that the virus exists in our ecosystem. Allergic alveolitis connected with the pigeon tick and the Dermanyssus gallinae mite also merits mention. In any case, where people have contact with birds or their droppings and secretions, special precautions should be taken. This way the negative effects of birds on human health can be minimised or eliminated

The variability of the ORF2a, ORF2b, ORF3, and ORF4 genes of the equine arteritis virus (EAV) was analysed during a seven year observation of persistent infection in a stallion of the Malopolska breed. A total of 11 semen samples were collected between 2004 and 2011. RNA of EAV isolates obtained from the semen of the stallion was amplified, sequenced, and compared with the sequences of other strains available in GenBank. Multiple nucleotide substitutions were found in sequences of the analysed regions, however, neither deletion nor insertions were detected. The highest number of point mutations (11-6 synonymous and 5 non-synonymous) were found in the ORF2b gene, and the lowest number of substitutions (6-5 synonymous and one non-synonymous) were found in the ORF2a gene. None of the identified mutations affected any of the glycosylation or phosphorylation sites of the minor EAV protein. Phylogenetic analysis of the ORF3 gene of EAV isolates showed that they grouped together within the cluster of European strains of EAV. Additionally, the ORF3 gene sequences of the isolates showed high (86.4% - 98.3%) similarity to the previously isolated Polish EAV strains.

The aim of the study was to analyse the prevalence of single-reactor (SR) pigs in Poland, to attempt an explanation for this phenomenon, and to assess whether the occurrence of SR pigs could create problems for a successful Aujeszky’s disease virus (ADV) eradication programme in Poland. A total of 6494 ADV gE antibody positive/doubtful sera were retested by gB ELISA and subsequently by virus neutralisation test (VNT) to confirm the results of the glycoprotein E (gE) ELISA. Amongst the serum samples tested, 5.9% could be classified as being taken from SR pigs, as was shown by gE ELISA positive/doubtful results, which were not confirmed by negative findings in gB ELISA and VNT. It means that the observed SR phenomenon was due to a false positive/doubtful reaction in gE ELISA. This finding was strongly supported by the fact that the serum samples were taken from the animals from herds without any previous or subsequent history of Aujeszky’s disease. The low percentage of SR pigs does not seem to create a big obstacle to a successful ADV eradication programme in Poland.

The aim of study was the preliminary evaluation of the occurrence of methicillin-resistant Staphylococcus aureus (MRSA) and extended spectrum ß-lactamases (ESBL) - producing Escherichia coli in 650 milk and inflammatory secretions from cows with clinical or subclinical mastitis. One millilitre of the sample was added to Mueller-Hinton broth supplemented with 6.5% NaCl, Tryptone Soya Broth with cefoxitin and aztreonam, and then to MRSA ID agar. Presumptive MRSA colonies were analysed for the presence of mecA gene. Parallel to MRSA identification, the samples were incubated in buffered peptone water, lauryl tryptose broth and McConkey agar supplemented with cefotaxim for ESBL-producing E. coli isolation. These bacteria were identified using API Rapid 32 E and the ability of ESBL production was initially established using disc test D68C and confirmed by MIC technique using Sensititre ESBL plates. The primers (blaCTX, blaTEM, blaSHV, and blaCMY-2-group) for the detection of some of the genes encoding ESBL production were used. The 45 strains of S. aureus with mecA gene and 41 strains of E. coli with blaTEM gene were detected.

The study evaluated the proliferative activity of immunocompetent cells in the jejunal and iliac lymph nodes of prepubertal female wild boars exposed to deoxynivalenol and zearalenone in naturally contaminated feed. The evaluation was performed with the use of the MTT assay and 2 mitogens: lipopolysaccharide (LPS) and concanavalin A. Intensified proliferative processes in T and B lymphocytes were revealed. The mitogenic activity of LPS was more expressed in the lymphocytes of both iliac and jejunal lymph nodes in comparison with the control group. Proliferative activity was higher in iliac lymph nodes than in jejunal lymph nodes. A reverse trend was observed in the percentage of live cells, which was higher in jejunal lymph nodes during the evaluation of lymphocyte proliferation.

The purpose of this study was to evaluate the efficacy of yeast glucomannan (YG) and sodium bentonite (SB) in reducing the toxicity in broilers fed a diet naturally contaminated with aflatoxin. Three hundred 7-day-old Ross 308 strain broilers were chosen and randomly assigned to 10 dietary treatments. Serum biochemical parameters and pathological changes in the liver were investigated at 42 d of age. Chickens fed a diet containing 250 ppb of aflatoxin displayed a decrease in uric acid, cholesterol, and triglycerides, and an increase in serum activities of AST and ALT when compared to control group. There were considerable gross and histopathological hepatic lesions (P < 0.05) in the form of small to moderate hydropic and/or fatty degeneration, bile duct hyperplasia, periportal fibrosis, cells infiltration, and congestion, in chickens fed the 250 ppb aflatoxincontaining diet. The addition of YG and SB to the aflatoxin-containing diet partially reduced the negative effects of aflatoxin. The 0.1% YG supplementation to the aflatoxin-contaminated diet significantly prevented the pathological effect of aflatoxin on serum biochemical parameters and liver, and was found to be more effective than other treatments.

The aim of the study was to estimate the current prevalence of E. multilocularis in selected populations of red foxes in Poland and to evaluate the changes in prevalence of this parasite by comparison with the results obtained in the same area during earlier surveillance. The investigations were performed in the area of four Polish provinces: 2 eastern/south-eastern (Lubelskie and Podkarpackie) and 2 south-western (Śląskie and Opolskie). Five hundred red foxes coming from the investigated areas were examined between 2013 and 2014 to estimate the current situation in selected provinces. Moreover, 550 red foxes from the same areas examined between 2007 and 2013 were used for comparison of differences in E. multilocularis prevalences in time. Intestines were examined with the use of the sedimentation and counting technique. Among 500 foxes examined in the current study, 118 were positive for E. multilocularis. There were differences in prevalence between individual provinces: Podkarpackie Province - 54.6%, Lubelskie Province - 18.9%, Śląskie Province - 11.7%, and Opolskie Province - 3.9%. Statistical analysis demonstrated that in most cases there were no differences in prevalence between the current results and the results from previous studies. Only in Opolskie Province was a statistically significant increase observed between 2010 and 2014. A stable degree of infection in the region with high prevalence of this parasite was demonstrated. However, a significant increase in the region with very low prevalence of E. multilocularis points out the necessity to monitor this infection during the coming years to control the progress of the disease

The purpose of this study was to assess the possible influence of beavers on the contamination of lake water with zoonotic parasites Giardia duodenalis and Cryptosporidium spp., with respect to the risk to human health. A total of 79 water samples were taken around the habitats of beavers from 14 localities situated in the recreational Masurian Lake District (north-eastern Poland). Water was sampled in the spring and autumn seasons, at different distances from beavers’ lodges (0-2, 10, 30, and 50 m). The samples were examined for the presence of (oo)cysts of zoonotic protozoa Giardia duodenalis and Cryptosporidium spp. by direct fluorescence assay (DFA) and by nested and real time PCR. By DFA, the presence of Giardia cysts was found in 36 samples (45.6%) and the presence of Cryptosporidium oocysts in 26 samples (32.9%). Numbers of Giardia cysts, Cryptosporidium oocysts, and summarised (oo)cysts of both parasites showed a significant variation depending on locality. The numbers of Giardia cysts significantly decreased with the distance from beavers’ lodges while the numbers of Cryptosporidium oocysts did not show such dependence. The amount of Giardia cysts in samples collected in spring was approximately 3 times higher than in autumn. Conversely, a larger number of Cryptosporidium oocysts were detected in samples collected in autumn than in spring. By PCR, Giardia DNA was found in 38 samples (48.1%) whereas DNA of Cryptosporidium was found in only 7 samples (8.9%). Eleven Giardia isolates were subjected to phylogenetic analysis by restriction fragment length polymorphism PCR or sequencing which evidenced their belonging to zoonotic assemblages: A (3 isolates) and B (8 isolates). In conclusion, water in the vicinity of beavers’ lodges in the tested region was markedly contaminated with (oo)cysts of Giardia duodenalis and Cryptosporidium spp., which confirms the potential role of beavers as a reservoir of these parasites and indicates a need for implementation of appropriate preventive measures to protect tourists’ health.

The aim of the study was the evaluation of the antimicrobial resistance of Enterococcus faecalis strains isolated from cattle, pig, and poultry meat. A test was performed on 111 strains using the minimum inhibitory concentration technique. The highest number of isolates (94 strains) were resistant to lincomycin, the second-highest resistance was to quinupristin/dalfopristin (88 strains), tetracycline followed (65 strains), and erythromycin resistance was also notable (40 strains). All isolates tested were sensitive to daptomycin, nitrofurantoine, and tigecycline, whereas only few strains were resistant to ciprofloxacin, gentamicin, penicillin, and vancomycin. The obtained results showed that meat may be a source of antimicrobial resistant enterococci which may be transferred to humans

The aim of the study was to identify the affinity of 10 Staphylococcus strains isolated from table chicken eggs to specific species. Preliminary analysis performed by API ID32 Staph test identified these strains as S. aureus, but they exhibited a negative reaction in the tube coagulase test. Thus, the analysed strains were initially characterised as Staphylococcus aureus-like (SAL). Further characterisation was performed by genotypic methods, using restriction fragment length polymorphism (RFLP) of the coagulase gene (coa) and sequencing of the gene rpoB. An attempt was also made to identify the isolated Staphylococcus strains by MALDI-TOF mass spectrometry. The results indicated that none of the strains tested belonged to the species S. aureus. The rpoB sequences of five isolates showed the highest sequence similarity to S. haemolyticus, three isolates to S. chromogenes, and one isolate to S. epidermidis. One strain (SAL4) remained unidentified in this analysis. The results obtained using mass spectrometry were comparable to those based on gene sequence analysis. Strain SAL4, which could not be identified by sequencing, was identified by MALDI-TOF as Staphylococcus chromogenes.

The paper presents the first results of a study on the contamination of honey produced in the Republic of Kazakhstan with C. botulinum spores known to pose a potential infection threat to infants. During microbiological analysis, culturing methods with TPGY, Willis-Hobbs agar, FAA agar connected with PCR, sequencing, and a mouse bioassay were used. The C. botulinum contamination rate of honey was relatively low as determined, at 0.91%. Nonetheless, the potential danger of the bacteria to childrens’ health should not be neglected

The aim of the study was to assess the microvessel density based on the analysis of the expression of the CD31, VEGF, and LIMS-1 proteins in canine mast cell tumours. The study was conducted on 60 mastocytomas; 16 cases were classified as the grade I, 26 as the grade II, and 18 as the grade III. Statistical analysis showed a positive correlation only between the grade of the tumour and the expression of LIMS-1. In conclusion, LIMS-1 could be successfully used as a prognostic endothelial cell marker in mast cell tumour. CD31 may be a useful marker, but further examinations are necessary. VEGF is not recommended

The objective of this study was to investigate the correlation between different histological types and grades of canine malignant mammary tumours, tumour cell proliferation and their angiogenic activity using immunohistochemical markers. Mammary tissue samples from 47 bitches with mammary cancer were evaluated. The expression of cellular proliferation marker Ki-67 and endothelial marker Von Willebrand’s factor (vWF) were immunohistochemically demonstrated. The tumours with the highest Ki-67 and vWF expressions were found to share similar histomorphological features. Simple solid carcinoma had the highest levels of Ki-67, vWF, and higher histological grade while complex carcinomas, osteosarcomas, and carcinosarcomas had the lowest ones. The differences between the expressions of Ki-67 and vWF in different tumour types were considered to be of great importance in determination of biological behaviour and prognosis of these tumours. This study is one of the few studies that evaluate these differences among the subtypes of malignant canine mammary tumours

Semen was collected from Polish Halfbred stallions. Twenty individuals from 3 to 4 years of age were selected for the study. At least one ejaculate from each stallion was collected and assessed. Sperm morphology was evaluated using Papanicolaou stain, SpermBlue^®, and a complex of eosin and gentian stain, whereas unstained slides were used as control samples. Morphometric measurements were performed on 100 randomly selected sperm heads in each ejaculate. The length, width, perimeter, and surface area of the sperm head were measured. The frequency of nuclear vacuoles was determined as well. Tygerberg’s strict criteria, which most precisely characterise the sperm head, were used in the morphological evaluation of the sperm. The results obtained indicate that in the case of staining with SpermBlue^® and with eosin+gentian complex, the morphometry of the sperm head seems to be most similar to that observed in unstained smears. It also seems that neither shrinking nor swelling is uniform in the various staining techniques (Papanicolaou stain). Moreover, it appears that in comparison with unstained sperm, all methods caused the width of the head to increase as a result of swelling.

This study was an attempt to evaluate sperm chromatin structure in the semen of insemination boars. Preparations of semen were stained with acridine orange, aniline blue, and chromomycin A3. Abnormal protamination occurred more frequently in young individuals whose sexual development was not yet complete, but may also be an individual trait. This possibility is important to factor into the decision regarding further exploitation of insemination boars. Thus a precise assessment of abnormalities in the protamination process would seem to be expedient as a tool supplementing morphological and molecular evaluation of semen. Disruptions in nucleoprotein structure can be treated as indicators of the biological value of sperm cells.

The aim of the study was to analyse sperm proteomes of ejaculates from Polish Large White (PLW) and Polish Landrace (PL) boars and to identify differences which putatively influence semen quality. Spermatozoa protein profiles were analysed by electrophoretic methods followed by selected techniques to evaluate semen quality on the following factors: sperm motility, lipid peroxidation levels (MDA production), ATP content, activities of superoxide dismutase (SOD) and catalase (CAT), total antioxidant status (TAS), and total oxidant status (TOS) of seminal plasma. A protein with an estimated molecular weight of 30 kDa was found in spermatozoa of selected ejaculates. Mass spectrometry demonstrated that this polypeptide is most similar to proacrosin binding protein (sp32). The presence of the protein was more frequently observed in sperm extracts obtained in spring-summer period. Ejaculates containing sp32-like protein demonstrated significantly higher spermatozoa motility, lower inhibition of MDA production by seminal plasma, and higher SOD activity in seminal plasma. Boar semen which included sp32-like protein also demonstrated lower ATP levels in spermatozoa as well as higher TAS and lower TOS of seminal plasma, though the differences were not statistically significant. Ejaculates from PLW boars, with sp32-like protein present in sperm, were characterised by significantly higher sperm motility, lower ATP content in spermatozoa, and higher TAS of seminal plasma. The diminished parameters of semen quality were observed in ejaculates from PL boars that also contained the discussed protein, but the differences were not statistically significant. These findings suggest that the presence of sp32-like protein in boar spermatozoa could influence semen quality

Twenty pigs of similar genetics (PIC) were used. Pigs were randomly divided into two groups: experimental (ENRO, n = 10) and control (C, n = 10). From day 0 to day 4, pigs from ENRO group received enrofloxacin at the recommended therapeutic dose. Pigs from C group received PBS instead of enrofloxacin. Blood samples were collected on days 0 (before antibiotic administration), 2, 4 (during antibiotic therapy), 9, and 13 of the study (after enrofloxacin administration). Haematological examination and flow cytometry were used to establish the relative and absolute counts of various leukocyte subsets. Lymphocyte subpopulations were measured by fluorochrom-labelled antibodies according to following definitions: CD3+ (T cells), CD21+ (B cells), CD4+CD8- (helper T cells, Th), CD4-CD8+ (cytolytic T cells, CLT), CD4+CD8+ (cytolytic and memory T cells). The present study revealed the modulating effect of enrofloxacin on the composition of circulating lymphocytes in pigs. Concentration and percentage of CD8+ cells decreased significantly after treatment with enrofloxacin and as a result the absolute CD4/CD8 ratio increased significantly as compared to control group (P < 0.05).These findings should prompt further studies on the practical significance of the results obtained in terms of clinical implications. In view of the results, it cannot be excluded that enrofloxacin may also have immunomodulatory effects on host response to infection.

The aim of the study was to evaluate the effect of flunixin and florfenicol administered in combination with vitamin E or C on selected leukocyte immune mechanisms and on the inflammatory process during the first few weeks in the feedlot. Fifty calves divided into 5 groups (n = 10) received florfenicol and flunixin with vitamin E or C. Blood was collected on the 1^st, 3^rd, 7^th, 14^th, 21^st, and 28^th d of the experiment. Intracellular metabolism (NBT), apoptosis, chemotaxis, susceptibility to M. haemolytica leukotoxin, and expression of β₂-integrins were determined in leukocytes. The symptoms of respiratory tract infection were observed in 40% of calves in control group, while in the other groups the morbidity rate ranged from 10% to 20%. Leukocytes showed decreased NBT, and the mean values for apoptosis ranged from 14% to 24%. The lowest percentage of apoptotic cells was observed in the calves that received florfenicol with flunixin and vitamins E and C. The chemotactic activity confirmed the significant inhibitory effect of the preparations on migration of the cells. A significant decrease (P ≤ 0.05) in the susceptibility of leukocytes to leukotoxin was noted in the group that received florfenicol and flunixin with vitamin E. Expression of β₂-integrin receptors was the lowest in calves receiving florfenicol with flunixin and vitamin E or C. The application of an antibiotic and a non-steroidal anti-inflammatory drug with antioxidants protected the leukocytes involved in defence against M. haemolytica virulence factors and effectively limited oxidative stress in the calves.

Horses (n = 20) were divided into 2 groups: oligofructose (OF)-induced equine laminitis group (group OF; n = 11) which received 10 g/kg b.w. of OF dissolved in 4 L water via nasogastric intubation, and control group (NS; n = 9) which received 4 L of saline. Blood was collected at 4 h intervals over 72 h study period and analysed by ELISA, kinetic limulus amoebocyte lysate assay, and glucose-oxidase methods. The level of insulin changed significantly in horses which received OF (P < 0.01); there was a significant negative correlation between the level of adiponectin and insulin over time. The results suggested that insulin may play an important role in the development of OF-induced equine laminitis by altering the level of endothelin-1 and nitric oxide.

The aim of this study was to assess the effect of thymus factor X (TFX-Jelfa) treatment in hypoestrogenic female rats for the purpose of decreasing skin impairments. Ovariectomised rats were used as a model. The histopathological analysis of the skin after TFX-Jelfa treatment demonstrated that the epidermis was thicker and more desquamated, without deep wrinkles or hypersecretion in comparison to the skin of animals only castrated and not treated with TFX-Jelfa. Collagen and elastic fibres were arranged more uniformly in the dermis and there were numerous fibroblasts, hair follicles, and small vessels. Ultrastructural analysis showed keratinocytes without degenerative changes and the proliferation of lymphatic and dendritic cells in the skin. The results indicate that thymus extracts can have beneficial effects on skin aging, which is often accompanied by hormonal perturbances.

The aim of the study was to identify magnetic resonance imaging (MRI) sequences that contribute to a quick and reliable diagnosis of brachial plexus tumours in dogs. The tumours were successfully diagnosed in 6 dogs by the MRI with the use of SE, FSE, STIR, Turbo 3 D, 3D HYCE, and GE sequences and the gadolinium contrast agent

The study was conducted on Polish Merino (143♀ and 39♂) and old-type Polish Merino sheep (176♀ and 61♂) in Brylewo flock (Wielkopolskie Province). The examined sheep were at the age of one year. Prion protein (PrP) genotype was determined in all animals. In both breeds four alleles (ARR, ARQ, AHQ, VRQ) were found. Highly significant effect of breed and insignificant impact of gender within breed was observed in regard to the frequency of occurrence of alleles and genotypes susceptible to classical scrapie in the sheep. Eight different PrP genotypes in Polish Merino and nine genotypes in old-type Polish Merino were identified. Very high frequency of ARR/ARR genotype in old-type Polish Merino and high frequency of ARR/ARQ genotype in Polish Merino were found, with relatively significant frequency of occurrence of the genotypes containing VRQ allele. In old-type Polish Merino, three animals (of both genders) had VRQ/VRQ genotype. Breeding work involving elimination of animals encoding valine at codon 136, and introduction of rams with ARR allele to the population increased the frequency of occurrence of ARR/ARR genotype and ARR allele in the population of old-type Polish Merino. To improve the distribution of the genotypes genetically resistant to scrapie in the flock of Polish Merino only rams with ARR/ARR genotype were left. This guarantees an increase in the frequency of occurrence of genotypes genetically resistant to scrapie in the offspring.