Volume 62 (2018): Issue 4 (December 2018)

Introduction: Traditionally, evolutionary analysis of equine influenza virus (EIV) is based on the HA gene. However, the specificity of the influenza virus enables the classification of viral strains into different phylogenetic groups, depending on the gene being analysed. The aim of the study was to analyse phylogenetic paths of EIV based on M gene with reference to the HA gene.

Material and Methods: M gene of Polish isolates has been sequenced and analysed along with all M sequences of EIV available in GenBank database. Phylogenetic analysis was performed using BioEdit, ClustalW, and MEGA7 softwares.

Results: The clustering of the strains isolated not only from Asia but also from Europe into one common Asian-like group of EIV was observed. Twelve nucleotide substitutions in the M gene of strains from the Asian-like group were crucial for the evolutionary analysis. We also observed homology in the M gene of the Asian-like and H7N7 strains.

Conclusions: M gene specific for the Asian-like group is present in strains recently isolated in Europe and Asia, which were classified previously in the Florida 2 clade based on HA. Therefore, Asian-like group does not seem to be assigned to a specific geographical region. Traces of H7N7 strains in more conservative genes like M of some contemporary EIV strains may indicate the link between the old phylogenetic group and recent H3N8 strains. Analysis of conservative genes may be more useful in tracking the direction of virus evolution than in the genes where the high variability rate may blur the original relationships.

Introduction:Staphylococcus pseudintermedius and Malassezia pachydermatis often cause skin diseases in dogs.

Material and Methods: An online survey was e-mailed to veterinary practices nationwide covering demographics, diagnosis methods, and oral and topical treatment options. Of the 740 surveys sent, 100 complete replies were obtained.

Results: The majority of clinicians were unaware of the existence of the International Society for Companion Animal Infectious Diseases guidelines or did not follow them (53%). Oral antibiotics were used universally for superficial bacterial folliculitis treatment, particularly amoxicillin-clavulanic acid (100%), cephalexin (94%), enrofloxacin (67%), or marbofloxacin (60%). For fold dermatitis (FD) and otitis externa (OE), oral antibiotics were also given as treatment in 88% and 82% of cases, respectively. Oral antifungals were often prescribed for generalised Malassezia dermatitis (85%), FD (70%), and OE (59%). S. pseudintermedius and M. pachydermatis were frequently treated topically, particularly with antibacterials or antifungals only, or a combination of antibacterials, antifungals, and glucocorticoids. Alternative options such as honey-based products were not frequently used.

Conclusion: Our survey suggests that oral antibiotics are overused by Portuguese clinicians despite the spread of antibiotic resistant S. pseudintermedius. Oral antibiotics and antifungals are commonly prescribed for skin conditions manageable with topical treatments.

Introduction: Genotype VI of avian avulavirus 1 (AAvV-1) has pigeons and doves as its reservoir and is often termed pigeon paramyxovirus type-1 (PPMV-1). The pathogenesis of PPMV-1 infections in poultry is largely obscure. It is known that PPMV-1 requires a series of passages in chickens before it becomes adapted to gallinaceous poultry.

Material and Methods: Changes in the genome of PPMV-1 were analysed after serial passages in specific pathogen free (SPF) chickens, using high-throughput sequencing. Additionally, histopathological lesions induced by PPMV-1 in experimentally inoculated pigeons, chickens, and turkeys were evaluated.

Results: Following six passages of PPMV-1 in chickens, 10 nonsynonymous substitutions were found including one (in the NP protein) which dominated the genetic pool of viral quasispecies. Histopathological changes induced by the post-passage PPMV-1 strain were more prominent than changes wrought by the pre-passaged PPMV-1 strain and the lesions were most intense in pigeons followed by chickens and turkeys.

Conclusion: PPMV-1 is highly adapted to pigeons and passaging through chickens results in the acquisition of novel amino acids in the polymerase complex, which may alter the pathogenic potential of the virus.

Introduction: The influence of feeding genetically modified MON 810 hybrid maize on the growth and haematological and biochemical indices of rats was tested.

Material and Methods: Two conventional (non-GM) and two test (MON 810) lines of maize were used in semi-purified diets at the level of 40% w/w. The non-GM I, MON 810 I, non-GM II, and MON 810 II maize lines were near-isogenic. A total of 40 male 6-week-old Wistar-derived rats were assigned to four equal feeding groups corresponding to the four maize lines for 16 weeks. Overall, health, body weight gain, clinical pathology parameters, gross changes, and appearance of tissues were compared between groups.

Results: There were no statistically significant differences in the weight gain or relative organ weights of rats, but there were some non diet-related histopathological changes in the liver, kidneys, and spleen. Except for creatinine level, no diet-related effects were observed in haematology or most of the biochemical indices. Transgenic DNA of MON 810 maize was not detected in the tissues or faeces nor in the DNA of E. coli isolated from the rectum digesta of rats given transgenic feeds. In our experiment, various metabolic indices of rats fed non-GM diets or genetically modified (MON 810) maize for 16 weeks were similar. No adverse nutrition-related health effects were detected.

Conclusion: MON 810 maize seems to be as safe as the conventional maize lines.

Introduction:Echinococcus granulosus is a zoonotic helminth of the Taeniidae family living in the small intestines of dogs. The hydatid cyst, which is the larval form of this parasite, is observed in sheep, goat, cattle, and many other organisms including humans. It causes a disease called cystic echinococcosis. Identification of strains of E. granulosus in dogs is critical in parasite control and eradication where possible. This study aims to determine the genotype of E. granulosus eggs and prevalence of this parasite in the faeces of dogs in the Van Province using the copro-PCR method.

Material and Methods: This study was conducted between 2015 and 2016 on the faeces obtained from 100 stray dogs from different parts of the Van Province. The coprological examination was conducted using the formalin-ether concentration method.

Results:Taeniidae eggs were found in 10 (10%) out of 100 faecal samples. E. granulosus was detected in 4 out of 10 of these (40%) infected samples. Sequence analysis of positive amplicons obtained from PCR showed that there were sheep strains (G1).

Conclusion: Dogs in Van area are primarily infected with the livestock genotype of E. granulosus, which is thought to be a potential zoonotic threat to humans.

Introduction: Immune-potentiating functions of Lactobacillus plantarum strains in the common carp were evaluated.

Material and Methods: Fourteen days of feeding fish dry diet supplemented with the bacteria provided parameters of nonspecific humoral immunity (lysozyme, ceruloplasmin, γ-globulin, total protein levels, and serum bactericidal activity) and cellular immunity (pinocytosis, respiratory burst activity, and potential killing activity of organ phagocytes), as well as the proliferative response of organ lymphocytes stimulated with mitogens. The resistance of fish to infection with Aeromonas hydrophila was also determined.

Results: Dietary supplementation with L. plantarum had a substantial influence on the activity of organ phagocytes, especially the potential killing activity of head kidney cells. A significant increase in the proliferative activity of LPS-stimulated B lymphocytes and in the levels of γ-globulins and total protein was observed. The supplemented diet conveyed higher resistance than the control diet as the cumulative fish mortalities after infection with A. hydrophila were 65% and 85%, respectively.

Conclusion: The results indicate that dietary supplementation with L. plantarum stimulates the antibacterial resistance of common carp and may reinforce defence against bacterial infections, but further studies need to be conducted.

Introduction: Toll-like receptors (TLRs) play a key role in the rapid activation of the innate immune response to a variety of pathogens. The aim of this study was to evaluate the effect of Trichinella spiralis infection on the level of expression of the tlr4 gene in mouse intestines during the intestinal phase of experimental trichinellosis.

Material and Methods: The experimental material consisted of the small and large intestines of BALB/c mice infected with Trichinella spiralis sampled at 4, 8, and 16 days post infection (dpi).

Results: A statistically significant increase was demonstrated in the tlr4 mRNA level isolated from the infected mice jejunum at 4, 8, and 16 dpi over the uninfected control. Moreover, at 4, 8, and 16 dpi in the jejunum of infected mice, a strong positive reaction for the presence of TLR4 protein compared with that of uninfected mice was observed.

Conclusion: Infection with T. spiralis changes the expression of the tlr4 gene in the small intestine of the mouse host.

Introduction: The extremely high genetic variation and the continuously emerging variants of foot-and-mouth disease virus (FMDV) of Southern African Territory (SAT) serotypes including SAT1, SAT2, and SAT3 make it necessary to develop a new RT-PCR for general use for monitoring viruses based on the updated genome information.

Material and Methods: A FMDV SAT-D8 one-step RT-PCR was established based on the 1D2A2B genes of the SAT serotype viruses with a multiplex primer set. FMDV A, O, C, and Asia 1 serotypes, other vesicular disease viruses, inactivated SAT viruses, and 125 bovine, ovine, caprine and porcine tissue samples collected from the Chinese mainland were included for evaluating the assay.

Results: The new RT-PCR was proven to be specific without cross-reactions with Eurasian FMDV, swine vesicular disease virus (SVDV), Seneca valley virus (SVV), or other common viral pathogens of cattle, sheep, goat, and pig. An around 257 bp-sized amplicon clearly appeared when the inactivated SAT viruses were detected. However, all 125 samples collected from FMDV-susceptible animals from the Chinese mainland which has not known SAT epidemics showed negative results.

Conclusions: A FMDV SAT-D8 one-step RT-PCR is a promising method for primary screening for FMDV SAT serotypes.

Introduction: Avian poxvirus infections are widespread in the domestic poultry population but are also reported in wild birds. In poultry, these infections cause significant economic losses, while wild birds may be a reservoir for poxvirus which affects breeding poultry. However, wild birds may also exhibit characteristic anatomopathological changes. This study concerns the infection of wild-living great tits (Parus major) with the avian poxvirus in Poland.

Material and Methods: Samples of internal organs and skin collected from great tits were homogenised and total cellular DNA was isolated. In PCR, the primers complementary to gene encoding the core protein 4b of the HP44 strain of fowl poxvirus (FPV) were used.

Results: After electrophoresis in 2% agarose gel, the PCR product of 578 bp characteristic for FPV was obtained in DNA samples isolated from skin lesions and the heart. The analysis of the nucleotide sequence of the virus strain showed 99% similarity to many poxviruses previously isolated from great tits and other free birds at various sites in the world.

Conclusions: This paper is the first clinically documented evidence obtained in laboratory conditions of avian poxvirus cases in great tits in Poland.

Introduction: Iron and molybdenum are essential trace elements for cell metabolism. They are involved in maintaining proper functions of enzymes, cell proliferation, and metabolism of DNA.

Material and Methods: BALB/3T3 and HepG2 cells were incubated with iron chloride or molybdenum trioxide at concentrations from 100 to 1,400 µM. The cells were also incubated in mixtures of iron chloride at 200 μM plus molybdenum trioxide at 1,000 μM or iron chloride at 1,000 μM plus molybdenum trioxide at 200 μM. Cell viability was determined with MTT reduction, LHD release, and NRU tests.

Results: A decrease in cell viability was observed after incubating both cell lines with iron chloride or molybdenum trioxide. In cells incubated with mixtures of these trace elements, a decrease in cell viability was observed, assessed by all the used assays.

Conclusions: Iron (III) and molybdenum (III) decrease cell viability in normal and cancer cells. A synergistic effect of the mixture of these elements was observed.

Introduction: Nickel and iron are very commonly occurring metals. Nickel is used in industry, but nowadays it is also used in medical biomaterials. Iron is an element necessary for cell metabolism and is used in diet supplements and biomaterials, whence it may be released along with nickel.

Material and Methods: BALB/3T3 and HepG2 cells were incubated with iron chloride or nickel chloride at concentrations ranging from 100 to 1,400 µM. The following mixtures were used: iron chloride 200 µM plus nickel chloride 1,000 µM, or iron chloride 1,000 µM plus nickel chloride 200 µM. The cell viability was determined with MTT, LHD, and NRU tests.

Results: A decrease in cell viability was observed after incubating the BALB/3T3 and HepG2 cells with iron chloride or nickel chloride. A synergistic effect was observed after iron chloride 1,000 μM plus nickel chloride 200 μM treatment in all assays. Moreover, the same effect was observed in the pair iron chloride 200 μM plus nickel chloride 1,000 μM in the LDH and NRU assays.

Conclusions: Iron (III) and nickel (II) decrease cell viability. Iron chloride at a concentration of 200 µM protects mitochondria from nickel chloride toxicity.

Introduction: Bisphenol A (BPA) is a substance widely used in industry for the production of polycarbonates and epoxy resins used in packaging and containers for beverages, contact lenses, compact discs (CDs), window panes, and many other elements. This compound belongs to the group of polyphenols and xenoestrogens commonly found in the human environment. What we know about BPA is still insufficient to enable us to protect our health against its adverse effects, and current knowledge of the influence of BPA on erythroblastic cell lines in bone marrow is rather fragmentary. The aim of the experiment was to assess the effect of two doses of BPA (0.05 mg/kg and 0.5 mg/kg b.w. per day) on myeloid haematopoiesis.

Material and Methods: During this experiment, the number of all types of cells in the erythroblastic cell line was evaluated in porcine bone marrow before and after BPA administration.

Results: The obtained results clearly indicate changes in haematopoietic activity of the bone marrow, which was demonstrated by a decrease in erythroblastic cell line production in both experimental groups. The haematological effects of the bone marrow changes were anaemia, caused by a number of erythrocytes which was depressed due to their immaturity, and a significant decrease in mean cellular volume in both groups.

Conclusion: The harmful effect of high and low doses of BPA on haematopoietic processes was proved.

Introduction: Exposure to zoonotic factors in veterinary practice is closely related to the nature of the work. The main aim of the study was to determine the risk of selected zoonotic infections among the occupational group of veterinarians in Poland.

Material and Methods: Blood samples of 373 veterinarians (162 males and 211 females) from 12 provinces of Poland were collected by the venipuncture of a forearm for serological tests. Commercial immunoenzymatic tests (ELISA) were used for detection of specific IgG antibodies to Echinococcus granulosus, IgM and IgG to Leptospira spp., and IgM, IgA, and I and II phase IgG to Coxiella burnetii. Enzyme-linked fluorescence assays (ELFA) were used to detect IgM and IgG antibodies to Toxoplasma gondii.

Results: Positive results were found in 209 (56.0%) veterinarians for at least one of the examined diseases. The overall proportion of participants found to have specific Toxoplasma gondii antibodies in the IgM and/or IgG assays amounted to 44.5%. The presence of Coxiella burnetii antibodies was found in 16 (4.3%) subjects, while Leptospira spp. antibodies were detected in 63 (16.9%) veterinarians. Among the 373 veterinarians examined, no Echinococcus granulosus antibodies were found.

Conclusion: Results of the study seem to indicate a slightly elevated risk of Toxoplasma gondii infection and a moderate risk of infection with Leptospira spp. and Coxiella burnetii in veterinarians.

Introduction: The aim of this study was to determine the predisposing effect of bovine respiratory syncytial virus (BRSV) on Pasteurella spp. infection in naturally-induced pneumonia in cattle by immunohistochemical labelling.

Material and Methods: Lungs of cattle slaughtered in the slaughterhouse were examined macroscopically, and 100 pneumonic samples were taken. The samples were fixed in 10% neutral formalin and embedded in paraffin by routine methods. Sections 5 μm in thickness were cut. The streptavidin-peroxidase method (ABC) was used to stain the sections for immuno-histochemical examination.

Results: BRSV antigens were found in the cytoplasm of epithelial cells of bronchi, bronchioles, and alveoles and within inflammatory cell debris and inflammatory exudate in bronchial lumens. Pasteurella spp. antigens were detected in the cytoplasm of the epithelial cells of bronchi and bronchioles, and in cells in the lumens of bronchi and bronchioles. Eleven cases were positive for only one pathogen (six for BRSV and five for Pasteurella spp.), while 35 cases were positive for 2 pathogens: BRSV plus P. multocida (n = 21) or M. haemolytica (n = 14).

Conclusion: The presence of high levels of BRSV in dual infections indicates that BSRV may be the main pneumonia-inducing agent and an important predisposing factor for the formation of Pasteurella spp. infections in cattle naturally afflicted with pneumonia.

Introduction: The study aimed to isolate thermophilic Campylobacter from chickens raised three rearing methods, determine its antimicrobial susceptibilities, and examine resistance-related genes by PCR.

Material and Methods: Cloacal swabs or intestinal contents were taken in Istanbul, Sakarya, and Izmir provinces. Chickens were from small village-based family-run businesses (n = 70), organically raised (n = 71), and conventionally raised broilers (n = 79). The samples were cultured on modified charcoal cefoperazone desoxycholate (mCCD) agar. Suspect isolates were identified with multiplex PCR (mPCR). As per EUCAST standards, MIC values were derived by broth microdilution for tetracycline, ciprofloxacin, nalidixic acid, kanamycin, gentamicin, and erythromycin in isolates of C. jejuni (n = 98) and C. coli (n = 83).

Results: In C. jejuni, 78.6% tetracycline, 87.8% ciprofloxacin, and 81.6% nalidixic acid resistance was detected, but none was to kanamycin, gentamicin, or erythromycin. In C. coli, 98.8% ciprofloxacin and 63.9% nalidixic acid resistance was detected, whereas resistance to nonquinolones was not observed. C257T (Thr-86-Ile) mutation in the gyrA gene of all phenotypically quinolone-resistant isolates was detected through a mismatch amplification mutation assay PCR (MAMA-PCR). It emerged that all isolates bore the tet (O) resistance gene.

Conclusion: Common tetracycline, nalidixic acid, and ciprofloxacin resistance exists in Campylobacter isolated from chickens raised three rearing methods.

Introduction: In calves, hyposelenosis degenerates skeletal muscles in different parts of the body. The extent of damage to muscle cells can be diagnosed by determining the activity of creatine kinase (CK), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH). The aim of this study was to analyse variations in the serum levels of LDH isoenzymes in calves with nutritional muscular dystrophy (NMD), to determine the applicability of this parameter for diagnosing NMD, and to describe the influence of hyposelenosis on total protein (TP), triglyceride (TG), and cholesterol (CHOL) levels.

Material and Methods: Two groups of calves (n = six animals per group) were used. After birth, control group calves (SC) were intramuscularly administered 10 ml of a preparation containing selenium (Se) and vitamin E, and experimental group animals (SE) that were not injected. Blood was collected after 5, 15, and 25 days, and the concentrations of Se, vitamin E, TP, TG, and CHOL and the activity of glutathione peroxidase (GSH-Px), CK, and LDH fractions were determined.

Results: Hypocholesterolaemia and elevated TG levels were found in SE group calves whose LDH fractions revealed a significant increase in LDH4 and LDH5 activity and a decrease in LDH1 activity when electrophoretically separated.

Conclusions: Nutritional muscular dystrophy is accompanied by hypocholesterolaemia and elevated TG levels caused by muscle lipolysis. LDH4 and LDH5 activity parameters assist early diagnosis of NMD in calves.

Introduction:Clostridium perfringens is commonly found in the gastrointestinal tract of animals and humans and continues to cause one of the most prevalent foodborne diseases in man.

Material and Methods: A total of 355 samples were examined for the occurrence of C. perfringens: rectal swabs from cattle, sheep, and goats, fresh stool samples from diarrhoea sufferers having been in contact with these animals, irrigation water and soil samples from the husbandry sites, and preharvesting fresh produce from farms irrigated with the sampled water. All samples were collected from Cairo and Giza governorates, Egypt. PCR analysis was carried out with positive isolates using the α-toxin gene. Sequence analysis of the gene of C. perfringens isolates was performed using the neighbour-joining approach. Bootstrap analysis was executed with 1,000 resamplings.

Results: 174 C. perfringens strains were isolated with a 49.01% prevalence. The highest prevalence of C. perfringens in apparently healthy animals was found in sheep (65.45%) followed by goats (58%), buffaloes (55%), and cattle (47.1%). Its prevalence in humans being in contact with these animals was 47.5%. The bacterium’s isolation from the soil and irrigation water was achieved in 40% and 31.7% of samples, respectively, posing a risk, particularly when the water and soil contact food in the field, shown by the fresh produce isolation of 40%. A significant relationship between the prevalence of C. perfringens in animal and environmental samples was identified (P < 0.05). A significant relationship was identified neither between animal species and C. perfringens prevalence, nor between the environmental source and C. perfringens prevalence (P > 0.05). All isolates were positive for the α-toxin gene by PCR. The sequence analysis and the phylogenetic relationship of the α-toxin genes from different samples revealed that C. perfringens from faeces of apparently healthy cattle, buffaloes, sheep, and goats is a significant threat in places where it can contaminate the soil and water. In addition, the sequence of C. perfringens from humans suffering from diarrhoea was found in the same cluster with the sequence from cows, goats, and sheep.

Conclusion: The role of apparently healthy animals in transmitting C. perfringens to humans, either through being in direct or indirect contact via water or soil in the cultivation of vegetables and fruits, was demonstrated.

Introduction: The objective was to determine the content of fatty acids in edible snail fat by snail species, collection site, and processing stage.

Material and Methods: The research material comprised 180 edible fat samples from the three genera of edible snails collected in Poland: free-living Helix pomatia (HP) and two cultivated Cornu subspecies: C. aspersa maxima (CAM) and C. aspersum aspersum (CAA). All snails came from the Greater Poland and Lower Silesian Provinces: HP from their natural habitat and CAM and CAA from heliciculture farms. The studies focused on the raw meat, cooked meat, and frozen meat processing stages. Fatty acid (FA) profiles were determined by the gas chromatography method.

Results:Helix pomatia fat showed a higher saturated fatty acid (SFA) content, whereas the fat of Cornu genus snails had a higher unsaturated fatty acid (UFA) component, i.e. monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA). Thermal processing of snail meat increased all the determined SFA and decreased all the PUFA values, and increased the content of C18:1, C20:1, and C22:1 acids in the MUFA group. The material collection site had limited impact on FA content as differences were noted only in levels of C18:1, C18:2 n6, and C20:5. The differences pertained only to the fat of farmed snails of the Cornu genus.

Conclusion: Due to the high content of UFA and a favourable ratio of n6:n3 acids and PUFA:SFA, snail fat can be regarded as nutritionally valuable.

Introduction: The aim of the study was the application and evaluation of real-time PCRs based on the fluorescence of SYBR Green I intercalating dye for the detection of three Bacillus anthracis genes in contaminated liver and blood samples. The goals for detection were rpoB gene as a chromosomal marker, pag gene located on plasmid pXO1, and capC gene located on plasmid pXO2.

Material and Methods: Five B. anthracis strains were used for the experiments. Additionally, single strains of other species of the genus Bacillus, i.e. B. cereus, B. brevis, B. subtilis, and B. megaterium, and strains of six other species were used for evaluation of the specificity of the tests. Three SYBR Green I real-time PCRs were conducted allowing confirmation of B. anthracis in the biological samples.

Results: The observation of amplification curves in real-time PCRs enabled the detection of the chromosomally encoded rpoB gene, pag gene, and capC gene of B. anthracis. The specificity of the tests was confirmed by estimation of the melting temperature of the PCR products. The sensitivity and linearity of the reactions were determined using regression coefficients. Strains of other microbial species did not reveal real-time PCR products.

Conclusion: All real-time PCRs for the detection of B. anthracis in biological samples demonstrated a significant sensitivity and high specificity.

Introduction:Coxiella (C.) burnetii, the aetiological agent of Q fever, is able to modulate the macrophage/T-lymphocyte axis in an infected organism and impair synthesis of monokines and lymphokines.

Material and Methods: The purpose of this research was to determine the levels of the cytokines that play a key role in the response to C. burnetii antigens (IL-1β, IL-2, IL-6, IL-10, IFN-γ and TNF-α) in the serum of animals originating from an infected herd prior to vaccination (day 0) and at 1, 7, and 21 days afterwards.

Results: The vaccination of animals did not affect the production of IL-6, IL-1β, or IL-2. The serum levels of these cytokines were too low to measure in most of the samples. The initial levels of TNFα, IFNγ, and IL-10 were higher in seropositive than in seronegative animals, although significant differences between seropositive shedders and seropositive nonshedders appeared only in the levels of IFNγ and IL-10. Additionally, the course of the post-vaccination response concerning these two cytokines was different among seronegative nonshedders, seropositive nonshedders, and seropositive shedders.

Conclusion: It seems that analysis of the IFNγ and IL-10 concentrations in animal blood serum may have some practical value in an assessment of the health status of seropositive animals and post-vaccination response.

Introduction: In the European Union, the use of thyreostatic drugs for fattening slaughter animals has been banned since 1981 under Council Directive 81/602/EEC. For protection of consumer health against unwanted residues and in compliance with Directive 96/23, each EU country must monitor thyreostats in samples of animal origin. This paper presents the results of research on thyreostatic residues carried out in Poland in 2011–2017.

Material and Methods: The material for testing was urine (n = 3,491), drinking water (n = 127), and muscle samples (n = 349) officially collected by Veterinary Sanitary Inspectors in slaughterhouses and farms throughout the country in accordance with the national residue control plan. The samples were examined for the presence of tapazole, thiouracil, methylthiouracil, propylthiouracil, and phenylthiouracil using liquid chromatography tandem mass spectrometry through an accredited method.

Results: In four bovine and three porcine urine samples, the permissible thiouracil concentration was exceeded. In one sample of porcine urine, methyl- and propylthiouracil were found. The presence of thiouracil and its derivatives in urine samples is most likely due to feeding animals diet containing cruciferous plants.

Conclusions: The results of research indicate that thyreostats are not used for anabolic purposes in slaughter animals in Poland.

Introduction: Highly pathogenic Asian H5-subtype avian influenza viruses have been found in poultry and wild birds worldwide since they were first detected in southern China in 1996. Extensive control efforts have not eradicated them. Vaccination prevents such viruses infecting poultry and reduces the number lost to compulsory slaughter. The study showed the efficacy of inactivated H5 vaccine from the H5N8 virus against highly pathogenic H5N8 and H5N6 avian influenza viruses in chickens.

Material and Methods: Reverse genetics constructed an H5 vaccine virus using the HA gene of the 2014 H5N8 avian influenza virus and the rest of the genes from A/PR/8/34 (H1N1). The vaccine viruses were grown in fertilised eggs, partially purified through a sucrose gradient, and inactivated with formalin. Chickens were immunised i.m. with 1 µg of oil-adjuvanted inactivated H5 antigens.

Results: Single dose H5 vaccine recipients were completely protected from lethal infections by homologous H5N8 avian influenza virus and shed no virus from the respiratory or intestinal tracts but were not protected from lethal infections by heterologous H5N6. When chickens were immunised with two doses and challenged with homologous H5N8 or heterologous H5N6, all survived and shed no virus.

Conclusion: Our results indicate that two-dose immunisations of chickens with H5 antigens with oil adjuvant are needed to provide broad protection against different highly pathogenic H5 avian influenza viruses.

Introduction: Avian reovirus (ARV) infections in poultry populations are reported worldwide. The reovirus belongs to the genus Orthoreovirus, family Reoviridae. The aim of the study was to evaluate the incidence of ARV infections in the poultry population based on diagnostic tests performed in 2010–2017.

Material and Methods: Samples of the liver and spleen were collected from sick birds suspected of ARV infection and sent for diagnostics. Isolation was performed in 5–7-day-old SPF chicken embryos infected into the yolk sac with homogenates of internal organs of sick birds. Four primer pairs were used to detect the σNS, σC, σA, and µA ARV RNA gene fragments. A nested PCR was used for the detection of the σNS and σC genes.

Results: In 2010–2017, ARV infection was found in birds from 81 flocks of broiler chickens and/or layers, 8 flocks of slaughter turkeys, and in 4 hatchery embryos at 17–20 days of incubation. The primers used in RT-PCR and nested PCR did not allow effective detection of ARV RNA in all virus-positive samples.

Conclusion: The problem of ARV infections in the poultry population in Poland still persist. The primers used for various ARV segments in RT-PCR and nested PCR did not allow effective detection of RNA in the visceral organs of sick birds. The presented results confirm the necessity of using classical diagnostic methods (isolation in chicken embryos, AGID).

Introduction: The ovarian follicular dynamics, vaginal electrical resistance (VER), progesterone (P4) and oestrogen (E2) profiles were investigated during the oestrous cycle in four indigenous ewes.

Material and Methods: Daily VER values were recorded with a heat detector. The follicles were observed and measured by trans-rectal ultrasonography. Blood was collected daily for hormonal profiles.

Results: A significant variation in VER values (P < 0.05) in oestrus by ewes and position in the sequence of cycles was observed. Trans-rectal ultrasonography of ovaries revealed the presence of 2–4 waves of follicular growth. Study of hormonal profiles by ELISA revealed a positive correlation between E2 concentration and development of follicles and a negative correlation between P4 concentration and their development. The concentrations of oestradiol increased in oestrus and then decreased to a basal level. Follicular growth was accompanied by a rise in the concentration of serum oestradiol. Inversely, when follicles received the stimulation for ovulation, concentration of progesterone started to fall, but after ovulation, it climbed back to its peak and remained at this state until next ovulatory follicle reached its maximum diameter.

Conclusion: This study could help to set up a manipulative reproductive technique for improving genetic values in indigenous sheep.