Volume 60 (2016): Issue 2 (June 2016)

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: A real-time RT-PCR method for identification and quantification of porcine epidemic diarrhoea virus (PEDV) strains in China was developed.

Material and Methods: Based on the conserved sequence of the PEDV nucleocapsid (N) gene, a primer pair and probe were designed to establish a TaqMan-MGB real-time RT-PCR assay for quantitative detection of the virus. The sequence was cloned into the pMD18-T vector and a series of diluted recombinant plasmids were used to generate a standard curve with an R2 value of 0.999.

Results: The developed quantitative PCR assay detected viral titres as low as 0.1 TCID₅₀ with high specificity and no cross-reaction with other porcine viruses (PoRV, TGEV, PRRSV, or CSFV). The intra-batch and inter-batch coefficients of variation were both less than 1%, which indicated good reproducibility. Thirty clinical diarrhoea samples obtained from pigs in Shanghai and Fujian were analysed using this quantitative PCR assay. Out of these samples, 93.3% were found to be PEDV positive.

Conclusion: This approach is suitable for clinical sample identification and pathogenesis studies.

Introduction: The aim of the study was to investigate the occurrence of goose haemorrhagic polyomavirus (GHPV) in wild birds inhabiting Poland.

Material and Methods: Samples from 508 birds of different species were obtained between 2010 and 2015. The internal organ sections were homogenised and then total cellular DNA was isolated. The study was performed by means of PCR assay using primers complementary to the VP1 gene of the GHPV.

Results: The presence of genetic material of GHPV was detected in 22 (4.33%) samples.

Conclusion: It was the first such study in Poland to emphasise the role of wild birds as a potential source of GHPV infection for farmed geese.

Introduction: In the present study, some of the commercial fish farms located in the Black Sea region of Turkey, were screened for bacteria between 2012 and 2014.

Material and Methods: The bacterial agents isolated from fish were identified by classical biochemical tests and the rapid diagnostic tests (API 20 E and API 20 Strep). All strains were further identified by sequencing of the 16S rRNA genes. The strains were also investigated for resistance to different antimicrobials by the disc diffusion method. Antibiotic resistance genes, including tetracycline (B), β-lactam (ampC, bla_TEM, bla_PSE), florfenicol (floR), erythromycine (ereA, ereB), sulphonamide (sulI, sulII), and trimethoprim (dhfr1) genes, were determined by the PCR method.

Results:Vibrio anguillarum, Vibrio fluvialis, Photobacterium damselae subsp. piscicida, Pseudomonas luteola, Lactococcus garvieae, Streptococcus iniae, Aeromonas hydrophila, and Yersinia ruckeri were isolated from marine and freshwater cultured fish. According to the results of disc diffusion, all isolates were sensitive to florfenicol, trimethoprim+sulfamethoxazole, oxitetracycline, and enrofloxacin, and resistant to lincomycin, penicillin G, and amoxicillin. Also, sulI, sulII, and floR resistance genes were detected in the bacteria.

Conclusion: The results of the study open up the opportunity to perform further investigations which could determine the possible role of ARGs in fish pathogens.

Introduction: The paper describes identification of Trichinella species isolated from wild boars (Sus scrofa) in the most popular hunting region of the West Pomeranian Province of Poland.

Material and Methods: The Trichinella larvae were identified by digestion method. For species identification of the larvae, multiplex PCR was used according to the European Reference Laboratory for Parasites Multiplex PCR protocol. The results were confirmed by molecular amplification of 5S rDNA gene and sequence analysis.

Results: Prevalence of 0.54% Trichinella–positive wild boars in the West Pomeranian Province was recorded. Examination of the larvae showed the occurrence of T. spiralis in 79 %, T. britovi in 16.5 %, mixed infection with T. spiralis/T. britovi in 3.5%, and T. pseudospiralis in 1.0% of the boars.

Conclusion: This is the first record of wild boar infected with non-encapsulated larvae of T. pseudospiralis in Poland. The species is very difficult to determine, especially using trichinoscopic method. The discovery of the larvae in the animals which may be intended for human consumption confirms that digestion technique should be the only method used for the inspection of meat, especially that from wild boars..

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: The current study was designed to detect Mycoplasma agalactiae (Ma), Mycoplasma mycoides subsp. capri (Mmc), Mycoplasma capricolum subsp. capricolum (Mcc) and Mycoplasma putrefaciens (Mp) in sheep and goats with clinical signs consistent with contagious agalactia.

Material and Methods: A total of 299 samples were collected from 55 mixed herds in Azarbaijan-e-Sharghi province, Iran. Samples were examined using PCR and culture methods.

Results: The findings showed that in 40 herds at least one sample was positive by PCR or culture method. Moreover, out of 274 sheep samples, 101 were proved to be positive using the PCR technique and 76 were found positive using the culture method. Out of 25 goat samples, 10 were found positive using PCR and 9 were positive through the culture method. Less than 20% of isolated mycoplasmas were Ma. Ma was detected from almost all studied regions in the province while Mmc, Mcc, and Mp were detected only in a very limited area that was deemed by the research group the mixed infection zone.

Conclusion: In vaccination or eradication projects, it would be more economical to focus on mixed infection zones. Further investigation on mixed infection zones could facilitate better understanding of contagious agalactia epidemiology.

Introduction: The aim of the study was to investigate changes in the serum levels of adiponectin and TNF-α, as well as insulin sensitivity, and to elucidate the possible relationship among the parameters and negative energy balance during the periparturient period of dairy cows.

Material and Methods: Thirty primiparous Holstein dairy cows were selected for the study. Blood samples were collected from each cow seven days before the expected calving date, on the calving day, and 7, 14, and 21 days after calving. Blood non-esterified fatty acids (NEFA), β-hydroxybutyric acid (BHBA), glucose, insulin, adiponectin, and TNF- α levels were measured. Revised Quantitative Insulin Sensitivity Check Index (rQUICKI) was calculated using data on NEFA, insulin, and glucose concentrations.

Results: When compared to prepartum levels, serum concentration of adiponectin significantly increased on day 21 postpartum. The rQUICKI increased and NEFA levels decreased on day 7 after parturition. Insulin and glucose levels decreased on days 7, 14, and 21 postpartum when compared with prepartum levels. BHBA levels decreased on day 21 and TNF- α concentration also decreased on days 7, 14, and 21 postpartum. Adiponectin levels positively correlated with NEFA during the preparturient period. Negative correlation was detected between adiponectin and rQUICKI on calving day and on 14^th day after parturition. TNF- α concentration positively correlated with glucose levels on day 7 prepartum and on 21^st day postpartum and with rQUICKI on 21^st day postpartum. Negative correlation was detected between adiponectin level and insulin sensitivity.

Conclusion: Based on the results of the study, we concluded that adiponectin could possibly increase insulin sensitivity when blood NEFA concentrations are elevated.

Introduction: The objective of this study was to describe a pharmacokinetic–pharmacodynamic (PK/PD) approach for determination of a rational dosage of ampicillin (AMP) and depletion of the antibiotic residues in milk after intramammary administration to cows.

Material and Methods: The cows came from different farms from the Lublin Province area. They (n = 9) received 5 g of the drug, containing 75 mg of AMP sodium in physiological solution, through a syringe tube by intramammary administration. Following single intramammary administration, the milk samples (5 mL) were collected after 2, 4, 6, 8, 10, 24, 36, 48, and 60 h. The liquid chromatography-mass spectrometry analysis was performed on the Agilent 1200 system connected to an AB Sciex API 4000™ mass spectrometer. The pharmacokinetic analysis of the concentrations of the antibiotic in milk was performed using software Phoenix^® WinNonlin^® 6.4. Calculations were made in non-compartmental (slopes, highest, amounts, and moments) and compartmental analysis.

Results: The pharmacokinetic characteristics of AMP after intramammary administration indicate rapid elimination of the drug from milk. The mean residence time had a several-fold lower value than the designated elimination half-life and amounts to only 3.4 h. The concentration of the drug in the milk dropped relatively quickly and the process was very dynamic.

Conclusion: The conducted research confirms the rationale of using the PK/PD model in order to verify the dosing regimen for other antibiotic groups and various indicators of the applied PK/PD model.

Introduction: Bovine postpartum metritis causes great losses. Mast cell (MC)-released mediators participate in uterine inflammation and immune response, but their role in postpartum metritis in cows has not been reported. This study investigated the effect of endometrial MC on the disorder.

Material and Methods: Ten dairy cows, at 6 to 10 days postpartum and with acute purulent metritis made up the experimental group, and 10 comparable healthy cows the control group. Endometrial histamine and IgE levels were determined by ELISA, and the MC particle state and expression of histamine H₁ (H₁R) and H₂ (H₂R) mRNA receptors were examined by transmission electron microscope and real-time quantitative PCR, respectively.

Results: Endometrial histamine and IgE levels were significantly higher in the experimental group. In the control group, homogenously distributed size-varied granules were seen in MC cytoplasm of endometrium of lamina propria. In the experimental group however, these showed degranulation with features of reduction. The level of H₁R mRNA was lower in the experimental group, but that of H₂R mRNA was higher.

Conclusion: The results suggest MC type I hypersensitivity characteristics during metritis, and histamine provocation of local inflammation. High expression of H₂R and low expression of H₁R inhibited the inflammatory response and prevented excessive uterine tissue damage.

Electroencephalography (EEG) is a non-invasive examination method for the assessment of functional central nervous system (CNS) disturbances. In human medicine it has a special importance as a diagnostic tool for epilepsy. Although many studies were done on the use of EEG for diagnostics of canine central nervous system disorders, the technique is still not applied routinely. The purpose of this paper was to review the use of the electroencephalography in canine neurological disorders of central nervous system diagnosis and assess the future perspectives of this technique in veterinary medicine.

Introduction: The aim of the study was to explore the effect of enrofloxacin on production of selected cytokines by porcine peripheral blood mononuclear cells (PBMCs).

Material and Methods: Twenty pigs (10 control and 10 experimental) were used in this research. Pigs from experimental group received enrofloxacin at therapeutic doses. Blood samples were collected before, during, and after treatment with enrofloxacin. PBMCs were incubated with or without lipopolysaccharide (LPS). Production of IL-6, IL-10, INF-γ, and TNF-α were determined by ELISA.

Results: Administration of enrofloxacin to healthy pigs for 5 d induced a transient reduction of the PBMCs response to LPS in terms of IL-6 and TNF-α secretion. The concentration of IL-6 returned to the day 0 level shortly after treatment, while TNF-α production remained reduced for 10 d after the treatment. The production of IL-10 was not affected by enrofloxacin. The level of IFN-γ was below the detection limit of the tests.

Conclusion: The results indicate that enrofloxacin administered in vivo in therapeutic doses has an immunomodulatory effect through its capacity to inhibit secretion of IL-6 and TNF-α by porcine PBMC stimulated by LPS.

Introduction: The main goal of the study was to investigate the effect of K_ATP channel modulators on development of oxidative stress in the heart of rats showing different resistance to hypoxia.

Material and Methods: The study has been performed on rats showing high- (HR) or low-resistance (LR) to hypoxia under modulators of ATP-sensitive potassium (K_ATP) channel opener pinacidil (0.06 mg/kg) and blocker glibenclamide (1 mg/kg) upon cobalt (Co) treatment (30 mg of cobalt chloride/kg b.w., 3 h). Changes in the oxidative stress parameters of the heart tissue, such as lipid peroxidation (LPO), level of oxidatively modified protein (OMP), and antioxidant defence system (superoxide dismutase - SOD, catalase -CAT, glutathione peroxidase - GPx, glutathione reductase - GR) as well as total antioxidant activity (TAA) were analysed.

Results: Co treatment caused a significant decrease in SOD and CAT activity in the heart of LR rats and GPx activity in HR rats. It also led to a decrease in OMP level in the heart of rats with HR in comparison with controls.

Conclusion: The obtained results suggest that individual resistance to hypoxia plays a crucial role in Co actions and provides evidence that the effects of K_ATP channel opener pinacidil in the heart are mediated through different pathways of the antioxidative system, depending on the individual resistance to hypoxia. Pinacidil exerts a protective effect on the heart tissue by preventing the LPO decrease and significantly reducing OMP levels, as well as increasing TTA in rats with LR.

Introduction: This article presents the analysis of the correlation between the category and health status of calves and the results of their rearing and levels of selected blood parameters.

Material and Methods: The study included 105 Polish Holstein-Friesian and beef (Limousine, Charolaise and Hereford) crossbred calves. Young bulls were purchased at the age of two to four weeks. The animals underwent quarantine, were dehorned, and 46 young bulls were castrated. The germ horns were removed by burning out. Castration was carried out with a bloodless method using a rubber band. The calves were kept in groups and fed a milk replacer administered via teats from automated milk-feeding stations. After the period of milk feeding, the calves were fed grass silage ad libitum and a concentrate at 2.5 kg/animal/day. The calves were weighed every two weeks. Blood for analyses was sampled at 43 d of age.

Results: After the rearing period finished at the age of six months, young bulls and steers had similar body weights (176.17 and 176.55 kg) and approximate average daily weight gains from birth (0.756 and 0.767 g/day). The healthy calves at six months of age weighed 180.47 kg, whereas the animals which at least once suffered from some diseases during rearing were lighter by approx. 30 kg (P ≤ 0.01). A statistically significant (P ≤ 0.01) difference was found for the count of red blood cells and white blood cells. In comparison with healthy individuals, the diseased animals had less RBC (8.33 and 9.42 10¹²/L respectively) and more WBC (27.03 and 12.26 10⁹/L respectively).

Conclusion: Castration of young bulls did not have any impact on the results of rearing and health status of the calves. The magnitude of the analysed parameters depended on the health status of the calves. Thus RBC and WBC parameters may be used to predict the health status of calves during rearing.

Introduction: The aim of this study was to investigate fluorescein use in the diagnosis of bladder ruptures in rabbits as an experimental model.

Material and Methods: The study was conducted on male New Zealand rabbits divided into a retrograde fluorescein group (n = 8) and an intravenous (IV) fluorescein group (n = 8). Following general anaesthesia, 10 mL of 10% fluorescein dye (sodium fluoresceine powder) was administered via ureterorenoscope to the bladder of the first group, and 0.5 mL of 10% fluorescein was administered intravenously to the second group. Then, the bladder was viewed through the cystoscope by urethral aspect. After experimental bladder perforation, groups were comparatively evaluated by paracentesis and laparotomy.

Results: Following IV injection of fluorescein dye, the bladder veins were stained green within 10 s and then fluorescein mixed with urine flowed into bladder lumen. The green fluid flow was observed in the abdominal cavity after the perforation of the bladder in both groups.

Conclusion: Fluorescein can be used as a marker in diagnosis of bladder ruptures. If there is no bleeding or intestinal content in the abdominal cavity, although a smoky yellow-green image is observed, bladder rupture can be suspected.

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 3^rd 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.