Introduction: The aim of the study was to determine the prevalence of intestinal helminths in red foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) in the Augustów Primeval Forest (north-eastern Poland), with particular regard to zoonotic parasites.

Introduction: The aim of the study was to determine the prevalence of intestinal helminths in red foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) in the Augustów Primeval Forest (north-eastern Poland), with particular regard to zoonotic parasites.Material and Methods: Intestines from 53 raccoon dogs and 66 red foxes were examined with the use of sedimentation and counting technique (SCT). Samples of faeces from 51 red foxes and 50 raccoon dogs were examined with the use of flotation method.Results: Parasitic helminths were found by SCT in 98.5% of red foxes and 96.2% of raccoon dogs. Both species were infected with: Alaria alata (93.9% and 94.3%, respectively), hookworms (68.2% and 83.0%), Apophallus spp. (7.6% and 15.1%), Mesocestoides spp. (57.6% and 24.5%), Taenia spp. (40.9% and 1.9%), and Toxocara/Toxascaris nematodes (33.3% 15.1%). Echinococcus multilocularis was detected only in red foxes (6.1%), but trematodes Echinostomatidae and nematodes Molineus spp. only in raccoon dogs (18.9% and 41.5%, respectively). Additionally, Capillaria spp. eggs were detected by flotation method in 78.4% of foxes and 20.0% of raccoon dogs.Conclusion: The study showed a very high percentage of red foxes and raccoon dogs infected with intestinal helminths in the Augustów Primeval Forest. Moreover, dangerous zoonotic parasites also were found, which should be taken into consideration in the assessment of infection risk for humans in this region.

African swine fever (ASF) is currently one of the most severe viral infections of domestic pigs, wild boars, and other hosts belonging to Suidae family. ASF is also considered as the most complex and devastating infectious and haemorrhagic disease of swine due to its severe socio-economic impact and transboundary character. ASF it is a notifiable disease and due to the lack of specific treatment and vaccine, the disease can be only limited by the administrative measures comprising wild boar hunting and stamping out of affected pigs. ASF occurred for the first time in Kenya in 1921 while in Europe (Portugal) the virus was detected at the end of the 1950s. In spite of successful eradication of this threat in a number of affected regions, the virus remains endemic in both feral and domestic pigs in Africa and Sardinia. The ‘new era’ of ASF started in 2007 after its re-introduction to Georgia. Following its intensive expansion, the virus spread to other Caucasian countries, including the territory of the Russian Federation. In 2014 the virus reached Ukraine, Belarus, and, consequently, European Union countries: Lithuania, Latvia, Estonia, and Poland. The occurrence of ASF in wild boars and pigs had a severe impact on both epidemiology and economy because of the national and international transport and trade consequences. Up to date, starting from the February 2014, eighty ASF cases in wild boar and three outbreaks in domestic pigs have been diagnosed. Taking into account the diverse rate of spread in Poland, this review aims to present and discuss the current state of knowledge on ASF including its epidemiology, pathology, transmission, and perspectives of control.

African swine fever (ASF) is currently one of the most severe viral infections of domestic pigs, wild boars, and other hosts belonging to Suidae family. ASF is also considered as the most complex and devastating infectious and haemorrhagic disease of swine due to its severe socio-economic impact and transboundary character. ASF it is a notifiable disease and due to the lack of specific treatment and vaccine, the disease can be only limited by the administrative measures comprising wild boar hunting and stamping out of affected pigs. ASF occurred for the first time in Kenya in 1921 while in Europe (Portugal) the virus was detected at the end of the 1950s. In spite of successful eradication of this threat in a number of affected regions, the virus remains endemic in both feral and domestic pigs in Africa and Sardinia. The ‘new era’ of ASF started in 2007 after its re-introduction to Georgia. Following its intensive expansion, the virus spread to other Caucasian countries, including the territory of the Russian Federation. In 2014 the virus reached Ukraine, Belarus, and, consequently, European Union countries: Lithuania, Latvia, Estonia, and Poland. The occurrence of ASF in wild boars and pigs had a severe impact on both epidemiology and economy because of the national and international transport and trade consequences. Up to date, starting from the February 2014, eighty ASF cases in wild boar and three outbreaks in domestic pigs have been diagnosed. Taking into account the diverse rate of spread in Poland, this review aims to present and discuss the current state of knowledge on ASF including its epidemiology, pathology, transmission, and perspectives of control.

Introduction: The aim of the study was to test the utility of Flinders Technology Associates filter paper (FTA® Cards) for molecular detection and storage of avian influenza virus (AIV). Material and Methods: There were two strains of AIV used in the study: low pathogenicity H7N1 and high pathogenicity H5N1 subtypes. Detection of viral material was conducted using molecular RT-PCR and rRT- PCR method. Results: The infectivity of LPAIV/H7N1 and HPAIV/H5N1 was completely inactivated within 1 h and 24 h after adsorption to FTA® Cards at room temperature, respectively. Viruses stored on FTA® Cards had detection limit approximately 1 log10 lower than live viruses. Viral RNA of both strains were detectable on the cards by rRT-PCR for a minimum of 150 d, irrespectively of storage temperatures (room temperature, -20ºC). RNA was also detected in all samples obtained from SPF chickens experimentally infected with HPAI/H5N1 on 3rd and 4th day post-infection (p.i.).

Introduction: The aim of the study was to test the utility of Flinders Technology Associates filter paper (FTA® Cards) for molecular detection and storage of avian influenza virus (AIV). Material and Methods: There were two strains of AIV used in the study: low pathogenicity H7N1 and high pathogenicity H5N1 subtypes. Detection of viral material was conducted using molecular RT-PCR and rRT- PCR method. Results: The infectivity of LPAIV/H7N1 and HPAIV/H5N1 was completely inactivated within 1 h and 24 h after adsorption to FTA® Cards at room temperature, respectively. Viruses stored on FTA® Cards had detection limit approximately 1 log₁₀ lower than live viruses. Viral RNA of both strains were detectable on the cards by rRT-PCR for a minimum of 150 d, irrespectively of storage temperatures (room temperature, -20ºC). RNA was also detected in all samples obtained from SPF chickens experimentally infected with HPAI/H5N1 on 3ʳᵈ and 4ᵗʰ day post-infection (p.i.). Conclusion: FTA® Cards enable safe and effective alternative transport of samples for molecular diagnosis of AIV.

Introduction: The present study is a comprehensive overview of the natural occurrence of 17β-oestradiol and testosterone in serum of cattle in Poland. Material and Methods: The serum samples (n = 826) were collected from cattle within five years. The samples were examined for the presence of oestradiol and testosterone using ELISA or gas chromatography with mass spectrometry. Results: In 98 samples (24%) 17β-oestradiol was detected above decision limits of applied methods, including five samples over the recommended concentration of 0.1 μg L-1. Of the serum samples taken from cows (≤18 months of age), 95 and 99 percentiles of the animals had 17β-oestradiol concentration below 0.027 and 0.086 μg L-1 and of samples from cows over 18 months of age - below 0.059 and 0.125 μg L-1 respectively. Calculated values for bulls (≤18 months of age) were 0.025 and 0.034 μg L-1 and for the animals older than 18 months of age - 0.035 and 0.041 μg L-1. The natural presence of testosterone was detected in 201 serum samples (48.7%). According to the obtained data, 95% and 99% of cows (≤18 months of age) serum samples had testosterone concentration below 0.05 and 0.23 μg L-1 and the animals over 18 months of age - 0.30 and 0.49 μg L-1, respectively. For bulls these values did not depend on the age of the animals and were in the ranges of 5 - 6.3 μg L-1 (95%) and 11.4 - 12.1 μg L-1 (99%). Conclusion: Our study showed that the threshold values for these hormones in plasma of cattle designated years ago are correct, but they need to be supplemented for animals older than 18 months.

Introduction: The present study is a comprehensive overview of the natural occurrence of 17β-oestradiol and testosterone in serum of cattle in Poland. Material and Methods: The serum samples (n = 826) were collected from cattle within five years. The samples were examined for the presence of oestradiol and testosterone using ELISA or gas chromatography with mass spectrometry. Results: In 98 samples (24%) 17β-oestradiol was detected above decision limits of applied methods, including five samples over the recommended concentration of 0.1 μg L⁻¹. Of the serum samples taken from cows (≤18 months of age), 95 and 99 percentiles of the animals had 17β-oestradiol concentration below 0.027 and 0.086 μg L⁻¹ and of samples from cows over 18 months of age - below 0.059 and 0.125 μg L⁻¹ respectively. Calculated values for bulls (≤18 months of age) were 0.025 and 0.034 μg L⁻¹ and for the animals older than 18 months of age - 0.035 and 0.041 μg L⁻¹. The natural presence of testosterone was detected in 201 serum samples (48.7%). According to the obtained data, 95% and 99% of cows (≤18 months of age) serum samples had testosterone concentration below 0.05 and 0.23 μg L⁻¹ and the animals over 18 months of age - 0.30 and 0.49 μg L⁻¹, respectively. For bulls these values did not depend on the age of the animals and were in the ranges of 5 - 6.3 μg L⁻¹ (95%) and 11.4 - 12.1 μg L⁻¹ (99%). Conclusion: Our study showed that the threshold values for these hormones in plasma of cattle designated years ago are correct, but they need to be supplemented for animals older than 18 months.

Introduction: The effects of Jin-Ying-Tang (JYT) on Toll-like Receptor 4 (TLR4) signalling transduction of lipopolysaccharide (LPS)-stimulated mouse mammary epithelial cells (MECs) in vitro were examined. Material and Methods: The cytotoxicity of JYT (0.06-62.50 mg/mL) on mouse MECs was determined by MTT assay. The MECs were co-cultured with LPS in the presence or absence of JYT (39.10 μg/mL, 391 μg/mL, 3910 μg/mL). The concentrations of interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α) in the culture supernatants were detected by ELISA. The mRNA expression of TLR4 and downstream TLR4 signalling molecules such as myeloid differentiation factor 88 (MyD88), tumour necrosis factor receptor associated factor 6 (TRAF-6), inhibitor κB (IκB), and nuclear factor κB inducing kinase (NIK) were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Results: The results showed that the IC50 of JYT on MECs was 12.25 mg/mL and JYT could significantly decrease the concentrations of IL-6 and TNF-α in LPS-stimulated MECs (P < 0.05). The mRNA expression of TLR4, MyD88, TRAF-6, IκB, and NIK was also significantly decreased when the LPS-stimulated MECs were cocultured at appropriate concentrations of JYT (P < 0.05, P < 0.01). Conclusion: These observations indicate a potential mechanism through which JYT attenuates the systemic inflammatory response to LPS-stimulated mouse mammary epithelial cells by inhibiting the activation of TLR4/MyD88/ TRAF-6/NIK pathway at the mRNA level.

Introduction: The effects of Jin-Ying-Tang (JYT) on Toll-like Receptor 4 (TLR4) signalling transduction of lipopolysaccharide (LPS)-stimulated mouse mammary epithelial cells (MECs) in vitro were examined. Material and Methods: The cytotoxicity of JYT (0.06-62.50 mg/mL) on mouse MECs was determined by MTT assay. The MECs were co-cultured with LPS in the presence or absence of JYT (39.10 μg/mL, 391 μg/mL, 3910 μg/mL). The concentrations of interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α) in the culture supernatants were detected by ELISA. The mRNA expression of TLR4 and downstream TLR4 signalling molecules such as myeloid differentiation factor 88 (MyD88), tumour necrosis factor receptor associated factor 6 (TRAF-6), inhibitor κB (IκB), and nuclear factor κB inducing kinase (NIK) were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Results: The results showed that the IC₅₀ of JYT on MECs was 12.25 mg/mL and JYT could significantly decrease the concentrations of IL-6 and TNF-α in LPS-stimulated MECs (P < 0.05). The mRNA expression of TLR4, MyD88, TRAF-6, IκB, and NIK was also significantly decreased when the LPS-stimulated MECs were cocultured at appropriate concentrations of JYT (P < 0.05, P < 0.01). Conclusion: These observations indicate a potential mechanism through which JYT attenuates the systemic inflammatory response to LPS-stimulated mouse mammary epithelial cells by inhibiting the activation of TLR4/MyD88/ TRAF-6/NIK pathway at the mRNA level.

Introduction: In the present study apoptosis was investigated in the cornu ammonis and cerebellum of 10 dogs naturally infected with rabies virus. Diagnosis of rabies was based on the results of fluorescent antibody staining and experimental inoculation.

Introduction: In the present study apoptosis was investigated in the cornu ammonis and cerebellum of 10 dogs naturally infected with rabies virus. Diagnosis of rabies was based on the results of fluorescent antibody staining and experimental inoculation.Material and Methods: The paraffin tissue sections were stained with haematoxylin and eosin, avidin-biotin complex peroxidase (ABC-P), and terminal deoxynucleotidyl transferase biotin-dUTP nick end-labelling (TUNEL) methods.Results: Histopathological examination revealed encephalomyelitis of varying severity and the presence of Negri bodies. Dense rabies antigens were determined in the motor neurons with ABC-P method. On the other hand, Bcl-2 protein and Bax protein gave positive reaction in seven and five cases, respectively. TUNEL staining demonstrated very marked apoptotic changes in the nuclei of neurons localised deep in the substantia alba of the cerebellum. Similar changes were also determined in perivascular mononuclear cells and glia cells within the substantia alba. No apoptopic changes were found in the motor neurons of the cornu ammonis.Conclusion: The absence of apoptotic changes in the neurons was considered to be the consequence of the necrotic changes that developed in these neurons.

Introduction: The aim of this study was to investigate the relationship between body condition score (BCS) determined on the dry-off day, calving day, and in the first month of lactation, its changes during the dry period and early lactation, and periparturient indices and fertility in high-producing dairy cows. Material and Methods: The experiment was conducted in two herds: A and B, located in Western Poland. The studies were conducted on 116 and 108 Polish Holstein-Friesian dairy cows respectively, with an average milk yield of >10 000 kg/305-day lactation. The experiment included the dry period (-56 d to the calving day), the calving day, and early lactation (from +1 to +56 d). The experimental factor was BCS (0 to 5-point scale). The BCS was performed by one person on day -56, on parturition day (in the first 12 h after calving) and on day 30 of lactation. Results: A decrease in BCS (≥-0.25) in herd A during the dry period accelerated the planned calving period by 7.3 d. In the group of cows with BCS <3.25 on the dry-off day, the lowest artificial insemination index (1.80), the shortest period of insemination services (25 d), and days open (87 d) were recorded. Moreover, cows with BCS < 3.25 at calving had the shortest days open (91 d). BCS >3.50 in the first month of lactation (30 d) resulted in the extension of uterine involution period (56 d). Improvement of BCS during the dry period shortened the anoestrus (60 d) in herd A and the period of insemination service (60 d) in herd B. However, in this group (IM BCS ≥ 0.25) of cows the day of the highest artificial insemination index (2.50) in herd B was analysed. Conclusion: The body condition on the dry-off day and at calving, as well as its deterioration in the first month of lactation, have a considerable effect on fertility indices in dairy cows, thus confirming the advisability of its regular monitoring during routine operations connected with the management of a dairy cattle herd.

Introduction: The aim of this study was to investigate the relationship between body condition score (BCS) determined on the dry-off day, calving day, and in the first month of lactation, its changes during the dry period and early lactation, and periparturient indices and fertility in high-producing dairy cows. Material and Methods: The experiment was conducted in two herds: A and B, located in Western Poland. The studies were conducted on 116 and 108 Polish Holstein-Friesian dairy cows respectively, with an average milk yield of >10 000 kg/305-day lactation. The experiment included the dry period (-56 d to the calving day), the calving day, and early lactation (from +1 to +56 d). The experimental factor was BCS (0 to 5-point scale). The BCS was performed by one person on day -56, on parturition day (in the first 12 h after calving) and on day 30 of lactation. Results: A decrease in BCS (≥-0.25) in herd A during the dry period accelerated the planned calving period by 7.3 d. In the group of cows with BCS <3.25 on the dry-off day, the lowest artificial insemination index (1.80), the shortest period of insemination services (25 d), and days open (87 d) were recorded. Moreover, cows with BCS < 3.25 at calving had the shortest days open (91 d). BCS >3.50 in the first month of lactation (30 d) resulted in the extension of uterine involution period (56 d). Improvement of BCS during the dry period shortened the anoestrus (60 d) in herd A and the period of insemination service (60 d) in herd B. However, in this group (IM BCS ≥ 0.25) of cows the day of the highest artificial insemination index (2.50) in herd B was analysed. Conclusion: The body condition on the dry-off day and at calving, as well as its deterioration in the first month of lactation, have a considerable effect on fertility indices in dairy cows, thus confirming the advisability of its regular monitoring during routine operations connected with the management of a dairy cattle herd.

In recent years a growing demand for ratite meat, including ostrich, emu, and rhea has been observed all over the world. However, consumers as well as the meat industry still have limited and scattered knowledge about this type of meat, especially in the case of emu and rhea. Thus, the aim of the present review is to provide information on technological and nutritional properties of ostrich, emu, and rhea meat, including carcass composition and yields, physicochemical characteristics, and nutritive value. Carcass yields and composition among ratites are comparable, with the exception of higher content of fat in emu. Ostrich, emu, and rhea meat is darker than beef and ratite meat acidification is closer to beef than to poultry. Ratite meat can be recognised as a dietetic product mainly because of its low level of fat, high content of polyunsaturated fatty acids (PUFA), favourable n6/n3 ratio, and high iron content in comparison with beef and chicken meat. Ratite meat is also rich in selenium, copper, vitamin B, and biologically active peptides such as creatine (emu) and anserine (ostrich), and has low content of sodium (ostrich). The abundance of bioactive compounds e.g. PUFA, makes ratite meat highly susceptible to oxidation and requires research concerning elaboration of innovative, intelligent packaging system for protection of nutritional and technological properties of this meat.

In recent years a growing demand for ratite meat, including ostrich, emu, and rhea has been observed all over the world. However, consumers as well as the meat industry still have limited and scattered knowledge about this type of meat, especially in the case of emu and rhea. Thus, the aim of the present review is to provide information on technological and nutritional properties of ostrich, emu, and rhea meat, including carcass composition and yields, physicochemical characteristics, and nutritive value. Carcass yields and composition among ratites are comparable, with the exception of higher content of fat in emu. Ostrich, emu, and rhea meat is darker than beef and ratite meat acidification is closer to beef than to poultry. Ratite meat can be recognised as a dietetic product mainly because of its low level of fat, high content of polyunsaturated fatty acids (PUFA), favourable n6/n3 ratio, and high iron content in comparison with beef and chicken meat. Ratite meat is also rich in selenium, copper, vitamin B, and biologically active peptides such as creatine (emu) and anserine (ostrich), and has low content of sodium (ostrich). The abundance of bioactive compounds e.g. PUFA, makes ratite meat highly susceptible to oxidation and requires research concerning elaboration of innovative, intelligent packaging system for protection of nutritional and technological properties of this meat.

Introduction: Chlamydia psittaci is a gram-negative obligate intracellular pathogen of birds. Poultry infections lead to economic losses and can be transmitted to humans. No vaccine is available and the bacterium-host cell interaction is not completely understood. Replicating bacteria cause pneumonia, but C. psittaci can also be non-replicating and persistent inside the cytoplasm of avian cells. RT-qPCR provides insight into the molecular pathogenesis of both active replicating and persistent Chlamydia psittaci in birds, but requires identification of stably expressed reference genes to avoid biases. Material and Methods: We investigated the expression stability of 10 C. psittaci candidate reference genes for gene expression analysis during normal growth and penicillin-induced persistence. C. psittaci Cal10 was cultured in HeLa229 and RNA was extracted. The expression level of each candidate was examined by RT-qPCR and Cq values were analysed using geNorm. Results: The genes tyrS, gidA, radA, and 16S rRNA ranked among the most stably expressed. The final selected reference genes differed according to the bacterial growth status (normal growth versus persistent status), and the time points selected during the duration of the normal chlamydial developmental cycle. Conclusion: The study data show the importance of systematic validation of reference genes to confirm their stability within the strains and under the conditions selected.

Introduction: Chlamydia psittaci is a gram-negative obligate intracellular pathogen of birds. Poultry infections lead to economic losses and can be transmitted to humans. No vaccine is available and the bacterium-host cell interaction is not completely understood. Replicating bacteria cause pneumonia, but C. psittaci can also be non-replicating and persistent inside the cytoplasm of avian cells. RT-qPCR provides insight into the molecular pathogenesis of both active replicating and persistent Chlamydia psittaci in birds, but requires identification of stably expressed reference genes to avoid biases. Material and Methods: We investigated the expression stability of 10 C. psittaci candidate reference genes for gene expression analysis during normal growth and penicillin-induced persistence. C. psittaci Cal10 was cultured in HeLa229 and RNA was extracted. The expression level of each candidate was examined by RT-qPCR and Cq values were analysed using geNorm. Results: The genes tyrS, gidA, radA, and 16S rRNA ranked among the most stably expressed. The final selected reference genes differed according to the bacterial growth status (normal growth versus persistent status), and the time points selected during the duration of the normal chlamydial developmental cycle. Conclusion: The study data show the importance of systematic validation of reference genes to confirm their stability within the strains and under the conditions selected.