Introduction: Ostrich characteristics include fast running, of which the probable enablers have been studied. Yet little research has taken place on one anatomical feature. It is mainly the special integuments on the ostrich foot which facilitate fast running on sand, because as point of direct sand contact they bear the whole weight and provide all the forward force. This study elucidates aspects of the integuments.Material and Methods: A stereo microscope, scanning electron microscope (SEM), and confocal scanning laser microscope were used to observe these integuments. Their surface structure was shown accurately in photographs. An SEM equipped with energy dispersive spectroscopy was used to check element contents of the upper and bottom areas and those on the lateral area of the 3ʳᵈ toe.Results: The content of some chemical elements on the upper area (Mg 2.04%, Si 0.18%, P 1.97%, Ca 0.59%, and S 0.69%) was higher than that of the bottom area (Mg 0.14%, Si 0.09%, P 0.10%, Ca 0.28%, and S 0.90%). Zinc was the particular element on the upper area, while sodium, chlorine, and potassium were the specific elements on the bottom area. The parts which must withstand different frictions contained different chemical compounds.Conclusion: The microscopic plane with layer-like structure and stripes may contribute to the wear-resistance of the papillae. The polygonal and prism structures are helpful to fix papillae in a firmer way.

Introduction: The aim of the study was to optimise and compare two multiplex PCR assays for the detection of Listeria spp. and Listeria monocytogenes in biological samples including the liver, brain, and blood. Material and Methods: Three strains of L. monocytogenes and single strains of each of the species: L. ivanovii, L. innocua, L. grayi, L. welshimeri, and L. seeligeri were used. Additionally, five other species of bacterium were used to evaluate the specificity of the tests. Results: Specific amplification products were obtained for both multiplex PCR assays, which confirmed the tested strains as Listeria spp. and L. monocytogenes, respectively. Isolates of other species did not yield PCR products. Conclusion: Both multiplex PCR assays proved to be significantly sensitive and highly-specific methods for the detection of Listeria strains.

Introduction: The aim of the study was to investigate the presence of human papillomaviruses (HPV), mouse mammary tumour virus (MMTV), Epstein-Barr virus (EBV), and human polyomavirus BK in canine mammary tumours (CMTs) and to correlate the results of histopathological classification with the results of virological examination. Material and Methods: Eighty CMTs and ten normal canine mammary gland samples were evaluated using histopathological methods and TaqMan real-time PCR analysis. Results: The results indicated that all mammary tumours and normal mammary tissue samples were negative for HPV16 and other HPV, EBV, human polyomavirus, and human mammary tumour virus strains. Conclusion: Further studies should be performed to investigate the existence of other strains of HPV, EBV, and human polyomavirus in CMTs.

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 study objective was to develop and evaluate a new TaqMan multiplex real-time PCR method for Salmonella, L. monocytogenes, and verotoxigenic Escherichia coli (VTEC) detection in slaughtered animal carcasses.Material and Methods: The procedure included an enrichment step, DNA extraction, and two multiplex real-time PCRs. The first PCR detected the invA and hly genes of Salmonella and L. monocytogenes respectively, the second the vtx1, vtx2, and eae genes of VTEC.Results: The validation of this method resulted in 100% relative sensitivity, specificity, and accuracy as compared to the reference ISO methods. The limit of detection per swab sample was established at 1 cfu for Salmonella and L. monocytogenes and 2 cfu for VTEC. The authors analysed 265 slaughterhouse-collected swabs from cattle, pig, and poultry carcasses. Among 125 from cattle, 51 were positive for VTEC, 29 for Salmonella, and 1 for L. monocytogenes. Among swabs from pig carcasses (n = 95), three, two, and one sample were positive for these pathogens respectively. None of the microorganisms tested for was identified in 45 samples of poultry origin.Conclusion: The obtained results showed that the method developed can rapidly identify the main bacterial pathogens that may contaminate carcasses of food-producing animals.

Introduction: The aim of the study was to determine the prevalence of respiratory capillariosis in red foxes (Vulpes vulpes) in some regions of Serbia.Material and Methods: The study was conducted on 102 foxes in six epizootiological regions of Serbia, during the hunting season between 2008 and 2012.Results: The presence of respiratory capillariosis in all tested epizootiological regions was confirmed. The E. aerophilus nematode was detected with overall prevalence of 49.02%. The diagnosis of E. aerophilus infection was confirmed by the determination of morphological characteristics of adult parasites found at necropsy and the trichurid egg types collected from the bronchial lavage and the content of the intestine.Conclusion: The presented results contribute to better understanding of the epidemiology of this nematodosis in Serbia. However, the high prevalence of capillaries in tested foxes, demonstrated in all explored areas, might suggest that foxes from other regions in Serbia may also be infected. The fact that domestic carnivores and humans can also be infected enhances the importance of the overall epidemiological status. To establish the relevant prevalence of respiratory capillariosis, further investigations and continous monitoring of parasitic fauna of carnivores are needed in the whole country.

Introduction: Mycoplasma bovis is one of the main pathogens involved in cattle pneumonia. Other mycoplasmas have also been directly implicated in respiratory diseases in cattle. The prevalence of different Mycoplasma spp. in cattle affected by respiratory diseases and molecular characteristics of M. bovis field strains were evaluated. Material and Methods: In total, 713 nasal swabs from 73 cattle herds were tested. The uvrC gene fragment was amplified by PCR and PCR products were sequenced. PCR/DGGE and RAPD were performed. Results: It was found that 39 (5.5%) samples were positive for M. bovis in the PCR and six field strains had point nucleotide mutations. Additionally, the phylogenetic analysis of 20 M. bovis field strains tested with RAPD showed two distinct groups of M. bovis strains sharing only 3.8% similarity. PCR/DGGE analysis demonstrated the presence of bacteria belonging to the Mollicutes class in 79.1% of DNA isolates. The isolates were identified as: Mycoplasma bovirhinis, M. dispar, M. bovis, M. canis, M. arginini, M. canadense, M. bovoculi, M. alkalescens, and Ureaplasma diversum. Conclusion: Different Mycoplasma spp. strains play a crucial role in inducing respiratory diseases in cattle.

Introduction: Hard antlers of deer are unique bioindicators of environmental metal pollutions, but sampling methods presented in the literature are inconsistent. Due to the specific growth pattern of antlers and their histological structure, sampling methods described in the literature were reviewed, the suitability of using mixed samples of both antler layers as element bioindicators was assessed, and the codified method of antler sampling used for bioindication was described. Material and Methods: Lead, cadmium, mercury, arsenic, copper, zinc, and iron in trabecular and cortical parts of hard antlers of red deer (Cervus elaphus) were determined using different methods of atomic absorption spectrometry (depending on the element). Results: Mean mercury content in trabecular bone (0.010 ±0.018 mg/kg) was 5 times higher than in cortical bone (0.002 ±0.003 mg/kg). Mean iron concentration was approximately 15 times higher in trabecular (239.83 ±130.15 mg/kg) than in cortical bone (16.17 ±16.44 mg/kg). Concentrations of other analysed elements did not differ statistically between antler layers. Conclusion: In mixed antler samples, concentrations of mercury and iron depend on the particular antler layer contents. This therefore warrants caution when comparing results across studies and specification of the sampling methodology of antlers is highly recommended.

Introduction: To identify novel pathways involved in the pathogenesis of ketosis, an isobaric tag for relative and absolute quantitation/mass spectrometry was used to define differences in protein expression profiles between healthy dairy cows and those with clinical or subclinical ketosis.Material and Methods: To define the novel pathways of ketosis in cattle, the differences in protein expression were analysed by bioinformatics. Go Ontology and Pathway analysis were carried out for enrich the role and pathway of the different expression proteins between healthy dairy cows and those with clinical or subclinical ketosis.Results: Differences were identified in 19 proteins, 16 of which were relatively up-regulated while the remaining 3 were relatively down-regulated. Sorbitol dehydrogenase (SORD) and glyceraldehyde-3-phosphate dehydrogenase (G3PD) were up-regulated in cattle with ketosis. SORD and G3PD promoted glycolysis. These mechanisms lead to pyruvic acid production increase and ketone body accumulation.Conclusion: The novel pathways of glycolysis provided new evidence for the research of ketosis.

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.