Introduction: The study was conducted to investigate the prevalence and genetic diversity of Chlamydia spp. in poultry in Poland and estimate possible transmission to humans. Material and Methods: Molecular diagnostic methods followed by sequencing and strain isolation were used on cloacal/faecal swabs collected from 182 apparently healthy poultry flocks including chickens, turkeys, ducks, and geese. Serum samples obtained from people exposed (study group) and non-exposed (control group) to birds were tested by complement fixation test to acquire data on Chlamydia spp. antibody level. Results: Overall, 15.9% of the tested flocks were Chlamydiaceae-positive and three Chlamydia spp. were identified. Predominant chlamydial agent found was C. gallinacea occurring in 65.5% of all positive poultry flocks and in 73.0% of positive chicken flocks. The sequences from four chicken flocks were assigned to C. abortus, whereas C. psittaci was confirmed in one duck and one goose flock. The analysis of ompA variable domains revealed at least nine genetic variants of C. gallinacea. Chlamydial antibodies were detected in 19.2% of human serum samples in the study group in comparison with 10.8% in the controls. Conclusion: The obtained results confirm that chlamydiae are common among chicken flocks in Poland with C. gallinacea as a dominant species. Moreover, the presence of C. abortus in chickens is reported here for the first time. Further investigation should focus on possible zoonotic transmission of C. gallinacea and C. abortus as well as potential pathogenic effects on birds’ health and poultry production.

Introduction: The study was conducted to investigate the prevalence and genetic diversity of Chlamydia spp. in poultry in Poland and estimate possible transmission to humans.

Introduction: Reindeer are adapted to long distance migration. This species can cope with variations in substrate, especially in ice and snow environment. However, few detailed studies about reindeer hoof are available. Thus this article describes the results of studies on macro- and micro-structures of reindeer hoof.Material and Methods: The gross anatomy of the reindeer hooves was examined. Stereo microscope (SM) and a scanning electron microscope (SEM) were used to observe four key selected positions of reindeer hooves. Moreover, element contents of the three selected positions of reindeer hooves were analysed using the SEM equipped with energy dispersive spectroscope.Results: Hoof bone structures were similar to other artiodactyl animals. In the microscopic analysis, the surfaces of the ungula sphere and ungula sole presented irregular laminated structure. Ungula edge surfaces were smooth and ungula cusp surfaces had unique features. Aside from C, O, and N, reindeer hooves contained such elements as S, Si, Fe, Al, and Ca. The content of the elements in different parts varied. Ti was the particular element in the ungula sole, and ungula edge lacked Mg and S which other parts contained.Conclusion: The macro- and micro-structures of the reindeer hooves showed high performance of skid and abrasion resistance. It is most probably essential to the long distance migration for the animals.

Introduction: Reindeer are adapted to long distance migration. This species can cope with variations in substrate, especially in ice and snow environment. However, few detailed studies about reindeer hoof are available. Thus this article describes the results of studies on macro- and micro-structures of reindeer hoof.

Introduction: Ornamental fish can suffer from different bacterial diseases. Among them the most prevalent are infections caused by Aeromonas, Shewanella, Citrobacter, Plesiomonas, Edwardsiella, and Pseudomonas. But there is a broad spectrum of rarely identified bacteria which may be causative agents of diseases. The aim of the study was to determine the species of bacteria pathogenic for fish which are prevalent in aquariums. Material and Methods: Bacteria were isolated from infected ornamental fish from pet shops and private aquariums in the Lublin region in 2015 and classified to species using MALDI-TOF MS. Results: A total of 182 isolates from ornamental fish were identified. The most frequent bacteria found in diseased fish were Aeromonas veronii (30.8% of total number of strains), A. hydrophila (18.7%), Shewanella putrefaciens (7.1%), Citrobacter freundii (7.1%), Pseudomonas spp. (7.1%), Shewanella baltica (4.9%), and Plesiomonas shigelloides (3.3%). Conclusion: Isolated bacterial species are facultative pathogens for fish and humans and may be isolated from fish without apparent symptoms of the disease.

Introduction: Ornamental fish can suffer from different bacterial diseases. Among them the most prevalent are infections caused by Aeromonas, Shewanella, Citrobacter, Plesiomonas, Edwardsiella, and Pseudomonas. But there is a broad spectrum of rarely identified bacteria which may be causative agents of diseases. The aim of the study was to determine the species of bacteria pathogenic for fish which are prevalent in aquariums.

Introduction: The purpose of the current study was to evaluate the blood glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) levels under seasonal variations in dairy cows during transition period, and to assess the relationship between chosen reproductive parameters, GSH-Px, and MDA. Material and Methods: Holstein cows calving in January were assigned into winter group (n = 42), while cows calving in August were assigned into summer group (n = 42). Blood samples were collected from the jugular vein 21, 14, and 7 days before calving, at calving (0 day), and 7, 14, and 21 days after calving. Reproductive parameters obtained from farm records were evaluated. Results: In both groups of cows, GSH-Px activity decreased from 21 days before calving to day 0, and it gradually continued to increase until 21 days after calving. GSH-Px activity was higher in winter group compared to summer group during the transition period (P < 0.05). MDA levels in both groups increased over time starting from 21 days before calving to 0 day, but it gradually decreased thereafter. MDA levels were higher in summer group compared to winter group during the transition periods (P < 0.05). Summer group of cows showed higher intervals of calving-to-oestrus, calving-to-conception, and higher insemination index (P < 0.01). Negative correlation was recorded between GSH-Px and MDA during all examination days (P < 0.01). MDA levels correlated with calving to conception interval on day 21 before calving and day 0 (P < 0.01) and insemination index on day 0 and 21 days after calving (P < 0.01). GSH-Px activity was negatively correlated with calving to conception interval on day 21 before calving, day 0, and 21 days (P < 0.01) after calving. Negative correlation on day 21 before calving and day 0 was also determined between GSH-Px and insemination index (P < 0.01). Conclusion: This study showed that blood oxidant and antioxidant levels have affected the fertility parameters in cows under seasonal variations.

Introduction: The purpose of the current study was to evaluate the blood glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) levels under seasonal variations in dairy cows during transition period, and to assess the relationship between chosen reproductive parameters, GSH-Px, and MDA.

Introduction: The main problem in determination of chloramphenicol in food of animal origin is a large number of matrices. The main target of this study was to create a method for determination and confirmation of chloramphenicol in products and food of animal origin. Material and Methods: Each 5 g matrix sample was mixed with 5 mL of water and 10 mL of acetonitrile/ethyl acetate, homogenised, and centrifuged. The organic layer was evaporated and redissolved in 6 mL of 4% NaCl. The extract was cleaned up by SPE technique. Chloramphenicol was analysed by LC-MS/MS in electrospray mode. Results: The procedure was validated according to the Commission Decision No. 2002/657/EC. The apparent recoveries were in the range of 92.1% to 107.1% with a repeatability less than 11.0% (4.4%-11.0%) and within-laboratory reproducibility below 13.6% (4.7%-13.6%). Conclusion: The method was successfully validated and proved to be efficient, precise, and useful for quantification of chloramphenicol in more than 20 different matrices.

Introduction: The main problem in determination of chloramphenicol in food of animal origin is a large number of matrices. The main target of this study was to create a method for determination and confirmation of chloramphenicol in products and food of animal origin. Material and Methods: Each 5 g matrix sample was mixed with 5 mL of water and 10 mL of acetonitrile/ethyl acetate, homogenised, and centrifuged. The organic layer was evaporated and redissolved in 6 mL of 4% NaCl. The extract was cleaned up by SPE technique. Chloramphenicol was analysed by LC-MS/MS in electrospray mode. Results: The procedure was validated according to the Commission Decision No. 2002/657/EC. The apparent recoveries were in the range of 92.1% to 107.1% with a repeatability less than 11.0% (4.4%-11.0%) and within-laboratory reproducibility below 13.6% (4.7%-13.6%). Conclusion: The method was successfully validated and proved to be efficient, precise, and useful for quantification of chloramphenicol in more than 20 different matrices.

Introduction: The paper presents the method of simultaneous determination of 10 illegal azo dyes in feed by ultra-high performance liquid chromatography coupled with tandem mass spectrometry technique. Material and Methods: The dyes were extracted with hexane, evaporated to dryness, and analysed. Separation was achieved in 7 min in a gradient elution using acetonitrile (A) and 0.1% formic acid (B) as a mobile phase. Results: The validation results showed the repeatability of the method, which was evaluated at three levels (50, 500, and 5,000 μg/kg). All the matrix calibration curves for the working ranges were linear (R² 0.9904 to 1.0), the repeatability was between 2.1% and 24%, and recoveries ranged from 77.9% to 120%. The LOD and LOQ were at 1-2 and 5-10 μg/kg for different dyes, respectively. Furthermore, the method was applied in the homogeneity tests of the in-house prepared feed containing Sudan I at the levels of 0.5, 5, and 50 mg/kg. Conclusions: A sensitive, selective, and fast multiresidue method was successfully developed and validated. Its robustness was confirmed by the analysis of an experimental feed containing Sudan I.

Introduction: The paper presents the method of simultaneous determination of 10 illegal azo dyes in feed by ultra-high performance liquid chromatography coupled with tandem mass spectrometry technique. Material and Methods: The dyes were extracted with hexane, evaporated to dryness, and analysed. Separation was achieved in 7 min in a gradient elution using acetonitrile (A) and 0.1% formic acid (B) as a mobile phase. Results: The validation results showed the repeatability of the method, which was evaluated at three levels (50, 500, and 5,000 μg/kg). All the matrix calibration curves for the working ranges were linear (R2 0.9904 to 1.0), the repeatability was between 2.1% and 24%, and recoveries ranged from 77.9% to 120%. The LOD and LOQ were at 1-2 and 5-10 μg/kg for different dyes, respectively. Furthermore, the method was applied in the homogeneity tests of the in-house prepared feed containing Sudan I at the levels of 0.5, 5, and 50 mg/kg. Conclusions: A sensitive, selective, and fast multiresidue method was successfully developed and validated. Its robustness was confirmed by the analysis of an experimental feed containing Sudan I.

Introduction: The prevention and control of Actinobacillus pleuropneumoniae in commercial production settings is based on serological monitoring. Enzyme-linked immunosorbent assays (ELISAs) have been developed to detect specific antibodies against a variety of A. pleuropneumoniae antigens, including long-chain lipopolysaccharides (LPS) and the ApxIV toxin, a repeats-in-toxin (RTX) exotoxin unique to A. pleuropneumoniae and produced by all serovars. The objective of this study was to describe ApxIV antibody responses in serum and oral fluid of pigs.Material and Methods: Four groups of pigs (six pigs per group) were inoculated with A. pleuropneumoniae serovars 1, 5, 7, or 12. Weekly serum samples and daily oral fluid samples were collected from individual pigs for 56 days post inoculation (DPI) and tested by LPS and ApxIV ELISAs. The ApxIV ELISA was run in three formats to detect immunlgobulins M, G, and A (IgM, IgG and IgA) while the LPS ELISA detected only IgG.Results: All pigs inoculated with A. pleuropneumoniae serovars 1 and 7 were LPS ELISA serum antibody positive from DPI 14 to 56. A transient and weak LPS ELISA antibody response was observed in pigs inoculated with serovar 5 and a single antibody positive pig was observed in serovar 12 at ≥35 DPI. Notably, ApxIV serum and oral fluid antibody responses in pig inoculated with serovars 1 and 7 reflected the patterns observed for LPS antibody, albeit with a 14 to 21 day delay.Conclusion: This work suggests that ELISAs based on ApxIV antibody detection in oral fluid samples could be effective in population monitoring for A. pleuropneumoniae.

Introduction: The prevention and control of Actinobacillus pleuropneumoniae in commercial production settings is based on serological monitoring. Enzyme-linked immunosorbent assays (ELISAs) have been developed to detect specific antibodies against a variety of A. pleuropneumoniae antigens, including long-chain lipopolysaccharides (LPS) and the ApxIV toxin, a repeats-in-toxin (RTX) exotoxin unique to A. pleuropneumoniae and produced by all serovars. The objective of this study was to describe ApxIV antibody responses in serum and oral fluid of pigs.

Tenderness is the most important characteristic of meat, determining consumer approval. There are numerous methods of its improvement, although of diverse effectiveness. addition of vitamin D₃ to the feed for a short period before slaughter (7–10 days) is one of the natural ways to enhance the tenderness. Vitamin D₃ is responsible for Ca²⁺ mobilisation in serum and increase in activity of proteolytic enzymes belonging to calpains, which results in significant improvement of beef tenderness and reduction of ageing time. The use of vitamin D₃ is an application tool determining tenderness improvement of beef with substantial reduction in processing costs. Moreover, shorter post mortem ageing process will exceed the retail display time, which will consequently reduce losses due to unsold meat being returned from shops to the manufacturers. Based on the results of studies conducted over the last 15 years, this paper presents the possibility and the effects of the use of vitamin D₃ to improve beef tenderness.

Tenderness is the most important characteristic of meat, determining consumer approval. There are numerous methods of its improvement, although of diverse effectiveness. addition of vitamin D3 to the feed for a short period before slaughter (7–10 days) is one of the natural ways to enhance the tenderness. Vitamin D3 is responsible for Ca2+ mobilisation in serum and increase in activity of proteolytic enzymes belonging to calpains, which results in significant improvement of beef tenderness and reduction of ageing time. The use of vitamin D3 is an application tool determining tenderness improvement of beef with substantial reduction in processing costs. Moreover, shorter post mortem ageing process will exceed the retail display time, which will consequently reduce losses due to unsold meat being returned from shops to the manufacturers. Based on the results of studies conducted over the last 15 years, this paper presents the possibility and the effects of the use of vitamin D3 to improve beef tenderness.

Introduction: Studies of anabolic hormone residues in the tissues of slaughter animals have been carried out in Poland for more than 25 years. During the period of 2011 to 2015, a total of 35 387 samples from different animal species were tested in the National Residue Control Programme for the presence of residues of compounds that cause hormonal effects, as listed in Annex 1 of Directive 96/23/EC. Material and Methods: The research was conducted in the National Reference Laboratory and eight regional laboratories in departments of veterinary hygiene located throughout the country. Urine, muscle tissue, serum, kidney fat, and drinking water were the targeted matrices. Test methods based on instrumental techniques such as gas and liquid chromatography coupled with mass spectrometry were applied, as well as enzyme-linked immunosorbent assays (ELISA). Results: The concentration of detected hormones exceeded the decision limits in 30 samples, the consequence of which was 41 non-compliances with current applicable criteria. The hormones found present pseudo-endogenous (nortestosterone and boldenone) only, while synthetic hormones were not identified. Conclusion: The non-compliant findings constitute a small percentage (0.085%) of the five-year analysis compilation. On this basis the related food produced in Poland can be accepted as safe for human consumption with regard to the hormone residues tested.

Introduction: Studies of anabolic hormone residues in the tissues of slaughter animals have been carried out in Poland for more than 25 years. During the period of 2011 to 2015, a total of 35 387 samples from different animal species were tested in the National Residue Control Programme for the presence of residues of compounds that cause hormonal effects, as listed in Annex 1 of Directive 96/23/EC.