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Mycobacteriosis are diseases caused by acid-fast mycobacteria other thanTissue samples of two male sitatunga antelopes (Tragelaphus spekii) were analyzed. Lymph node and lung samples were subjected to anatomical examination and Ziehl-Neelsen staining. Real-time PCR was performed to confirm or rule out tuberculosis mycobacteria infection. In order to isolate the bacterial strain, tissue samples were inoculated on both solid and liquid media. HainLifescience CM tests, mass spectrometry and New Generation Sequencing were used to determine the mycobacterial species.Results showed that atypical mycobacteria are responsible for the antelope disease. The results of the HainLifescience CM test and mass spectrometry indicated that the mycobacterium responsible for causing mycobacteriosis wasThe sitatunga antelope is an animal susceptible to infection by

Introduction The article describes the occurrence and phylogenetic relationship of Salmonella isolates found in subcutaneous abscesses of leopard geckos. The aim of the study was to determine the cause of the abscesses and to characterise isolated Salmonella strains. Material and Methods Samples of abscesses from five animals and internal organs (lungs, liver, and gut) of three of them were tested for Salmonella according to the PN-EN ISO 6579:2002/A1:2007 standard. The antimicrobial resistance was evaluated by minimal inhibitory concentrations and the genetic similarity of the isolates was assessed with pulsed field gel electrophoresis (PFGE). Results In total, seventeen Salmonella isolates belonging to five different serovars were found to be susceptible to all tested antimicrobials except streptomycin. The serovars were S. Hadar, S. Fluntern, S. Tennessee, S. enterica subsp. salamae 55:k:z39, and S. Kentucky. Up to three serovars from different organs were isolated from the same individual. In two geckos, Salmonella were detected in the lungs. In three serovars, XbaI-PFGE typing revealed indistinguishable isolates from organs and abscesses. Conclusion Multiple Salmonella serovars might be involved in abscess formation and infections. The occurrence of the same PFGE profiles of the isolates may testify to the role of opportunistic organisms in causing infection.

Introduction Since 2009, Poland has been recognised as a country officially free of bovine tuberculosis (bTB), although in each year of the last five there were from 8 to 18 outbreaks of the disease. In 2008–2016, the largest number of cattle infected with bovine mycobacteria were eliminated in the Masovian Province (the central region of Poland) and the largest number of outbreaks of this zoonosis were recorded in this area. The close proximity of farms where bTB was found led to the suspicion that tuberculosis could have been transmitted between the affected herds. The aim of the study was the molecular characterisation of the pertinent M. bovis/caprae strains and determination of the epidemiological relationship of various bTB outbreaks. Material and Methods The material for microbiological tests came from 119 cattle (Bos taurus) from nine herds located in five provinces, neighbouring the Masovian Province. Results Laboratory tests of tissue material gave results confirming tuberculosis in 54 (45%) animals. All strains belonged to the Mycobacterium bovis species. A two-step analysis of genetic affinity allowed 50 strains to be identified as phylogenetically closely related and separated between three genetic clusters consisting of 2 to 27 strains. Conclusion Based on the results of genotyping, bTB outbreaks were found in three herds, and three transmission chains were identified among these herds.

The most numerous group of bacteria in the genus

Introduction and objective Bovine tuberculosis (bTB, bovine TB) is caused by mycobacteria which are grouped within the MTBC. TB in animals is a highly infectious and progressive disease which can be transmitted to humans. Since 2009, Poland has gained official bTB-free status. Despite the official fact of bTB-free status, a dozen bTB outbreaks are still noted each year. Since 2000 in Poland, every year 1/5 of the national herd is subject to intradermal skin TB testing to control the bTB outbreaks in the cattle population. Application, with 5-year intervals between each government-funded skin test, undoubtedly resulted in financial savings. However it also seems to have caused several adverse and worrying events, e.g. an increase in the number of reactors detected and removed from a single tested herd. The objective of this study was the examination of 898 cattle imputed with bTB infection in Poland between 2008-2012. Material and methods The study concerned a potential epidemic outbreak with suspected bTB transmission. 20 cows came from 3 herds in the same county located in the same province in southern Poland. Results 134 MTBC strains were identified. In MIRU-VNTR, all isolates showed the same genetic pattern 322532243421232. Based on molecular investigation, the characteristics of M. bovis strains isolated from cattle from 3 different herds confirmed the common source of this zoonotic disease. Conclusions Although not bacteriologically proven, everything points to the fact that humans were the vector of bovine tuberculosis transmission between herds. This finding confirms transmission between 3 cattle herds in the Malopolskie Province in southern Poland (Podhale). The outbreak of tuberculosis in animals finally compromised public health.

Introduction Bovine tuberculosis is a chronic contagious disease caused by Mycobacterium bovis or Mycobacterium caprae. Before widespread action conducted in Poland between 1959-1975 to combat bovine tuberculosis (BTB), about 40% of all tuberculosis cases in pigs was caused by the bovine bacillus. At the present time, correctly carried out, long-term control of cattle has resulted in cases of bovine tuberculosis in pigs and humans being extremely rare and sporadic. In pigs, tuberculosis is most often caught in a slaughterhouse during slaughter. Material and methods Samples came from pigs kept on the farm. Traditional bacteriological methods on solid media (Stonebrink, LJ with pyruvate) supported by the semi-automatic, liquid indicative culture method (MGIT) and PCR test were applied in targeted studies. The GenoType Mycobacterium MTBC and CM tests (Hain Lifescience, Germany) were used to additionally confirm that isolated strains classification was used. Results Strains of mycobacteria were isolated from all examined pigs. Mycobacterium bovis was determined by real time PCR and Hain Genotype methods. Conclusions In order to effectively fight against BTB, all animals on farms should be tested, regardless of species, while the milk of suspected cows should be utilized without being used for feed. It is important to adapt the current legal regulations to the current epidemiological situation.

Reptiles are considered a reservoir of a variety of Salmonella (S.) serovars. Nevertheless, due to a lack of large-scale research, the importance of Reptilia as a Salmonella vector still remains not completely recognized. A total of 731 samples collected from reptiles and their environment were tested. The aim of the study was to assess the prevalence of Salmonella in exotic reptiles kept in Poland and to confirm Salmonella contamination of the environment after reptile exhibitions. The study included Salmonella isolation and identification, followed by epidemiological analysis of the antimicrobial resistance of the isolates. Implementation of a pathway additional to the standard Salmonella isolation protocol led to a 21% increase in the Salmonella serovars detection rate. The study showed a high occurrence of Salmonella, being the highest at 92.2% in snakes, followed by lizards (83.7%) and turtles (60.0%). The pathogen was also found in 81.2% of swabs taken from table and floor surfaces after reptile exhibitions and in two out of three egg samples. A total of 918 Salmonella strains belonging to 207 serovars and serological variants were obtained. We have noted the serovars considered important with respect to public health, i.e., S. Enteritidis, S. Typhimurium, and S. Kentucky. The study proves that exotic reptiles in Poland are a relevant reservoir of Salmonella.

The most numerous group of bacteria in the genus Mycobacterium is the nontuberculous mycobacteria. Currently, over 200 species of bacteria have been classified as belonging to this group, of which approximately 30 are pathogenic to humans and animals. Mycobacterium kansasii complex numbers among these pathogenic species. The submandibular lymph nodes of a wild boar shot by a hunter were examined in order to confirm or exclude infection with bacteria of the genus Mycobacterium. In culture, a bacterial isolate was obtained after 12 days of incubation on Petragnani and Stonebrink media. A multiplex PCR clearly indicated that the isolate was a nontuberculous mycobacterium. The results of species identification attempts via both molecular biology methods and mass spectrometry confirmed that the isolated strain belonged to MKC. The described case of a wild boar infection with MKC is the first documented case in Poland and only the second in Europe, and in confirming the presence of this pathogen among free-living animals, this report implies that MKC is of great concern. Our research elucidates some specifics of wild boar mycobacteriosis and may be used to instill awareness in the public of the dangers of dressing hunt prey or consuming its meat in ignorance of safe procedures, which can contribute to the transmission of the pathogen to humans.

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: Tuberculosis is a highly infectious disease affecting humans and animals. It is caused by the Mycobacterium tuberculosis complex (MTBC) – Mycobacterium bovis and Mycobacterium caprae, which are aetiological factors of bovine tuberculosis (bTB). In Poland, the bTB eradication programme exists. Animals diagnosed with tuberculosis are in the majority of cases not treated, but removed from their herd and then sanitary slaughtered. Material and Methods: In total, 134 MTBC strains isolated from cattle in Poland were subjected to microbiological analysis. The resistance phenotype was tested for first-line antimycobacterial drugs used in tuberculosis treatment in humans: streptomycin, isoniazid, rifampicin, ethambutol, and pyrazinamide. The strains were isolated from tissues collected post mortem, so the test for drug resistance fulfilled only epidemiological criterion. Results: The analysis of drug-resistance of MTBC strains revealed that strains classified as M. bovis were susceptible to 4 antimycobacterial drugs: isoniazid, rifampicin, streptomycin, and ethambutol, and resistant to pyrazynamide. The strains classified as M. caprae were sensitive to all tested drugs. Conclusion: The results indicate that despite enormously dynamic changes in mycobacterial phenotype, Polish strains of MTBC isolated from cattle have not acquired environmental resistance. The strains classified as M. bovis are characterised by natural resistance to pyrazinamide, which is typical for this species.

Introduction Bovine tuberculosis (bTB) is an infectious disease that occurs in many species of both domestic and free-ranging animals, as well as animals kept in zoos. According to the Polish regulations, cattle tuberculosis are slaughtered and microbiological examinations are performed, the rest of animal species can be treated and laboratory diagnostics are not obligatory. Case report The presented case concerns two male giraffes which were purchased by the zoo and united with a third male. After a year, the oldest male died. Post mortem examinations confirmed generalized tuberculosis. After a further six months, the second male was euthanized after suffering great pain. The material for the study of drug resistance was a swab from the nose, obtained ante mortem from the third male. Attempted treatments did not produce the expected results. Genotyping allowed the exclusion of a common source of transmission. Conclusions The final effect of the anti-tuberculosis therapy in the male giraffe raises the question whether the research team should have undertaken the treatment of the animal with active tuberculosis.

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.

The accurate identification of Taylorella equigenitalis strains is essential to improve worldwide prevention and control strategies for contagious equine metritis (CEM). This study compared 367 worldwide equine strains using multilocus sequence typing according to the geographical origin, isolation year and equine breed. The strains were divided into 49 sequence types (STs), including 10 described for the first time. Three major and three minor clonal complexes (CCs), and 11 singletons, were identified. The genetic heterogeneity was low (0.13 STs/strain) despite the wide diversity of geographical origins (n = 16), isolation years (1977 to 2018) and equine breeds (n = 18). It was highest outside Europe and in the 1977-1997 period; current major STs and CCs already existed before 1998. Previous data associated the major CC1 with the first CEM outbreaks in 1977-1978 in the United Kingdom, Australia and the United States, and revealed its circulation in France. Our study confirms its circulation in France over a longer period of time (1992-2018) and its distribution in Spain and Germany but not throughout Europe. In addition to CC1, relationships between non-European and European countries were observed only through ST4, ST17 and ST30. Within Europe, several STs emerged with cross-border circulation, in particular ST16 and ST46 from the major complexes CC2 and CC8. These results constitute a baseline for monitoring the spread of CEM outbreaks. A retrospective analysis of a higher number of strains isolated worldwide between 1977 and the early 2000s would be helpful to obtain an exhaustive picture of the original CEM situation.

The aim of the study was the application and comparison of real-time PCR methods based on the fluorescence of SYBR Green I intercalating dye and TaqMan probes for the detection of the 23S rDNA gene of Listeria spp. and the hlyA gene of Listeria monocytogenes. Five 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 for the experiments. Additionally, five strains of other species of bacteria were used for evaluation of the specificity of the tests. QuantiTect SYBR Green PCR and QuantiTect Probe PCR kits were selected for the study. In the first stage of the study, SYBR Green I real-time PCRs were performed under several methods, the first one allowing detection of the 23S rDNA gene and the remainder based on the amplification of the hlyA gene. In the next part, three varied in method TaqMan probe-based real-time PCRs allowing confirmation of strains belonging to Listeria spp. and L. monocytogenes were conducted. The observation of amplification curves in real-time PCR methods enabled the detection of both genes, and these methods demonstrated a significant sensitivity and high specificity. A high regression coefficient of 0.99 was found for all reactions. Specific amplification products were obtained for the 23S rDNA and hlyA genes, which confirmed the tested strains as Listeria spp. and L. monocytogenes respectively. Isolates of other microbial species did not yield real-time PCR products.

The paper describes avian tuberculosis in a captive bred cassowary. A two-and-a-half-year-old bird was obtained by a Polish zoo in 2010 from the Netherlands under conditions compliant with the recommendations of the European Association of Zoos and Aquaria. Despite being of small size for the age, the bird appeared healthy and showed no signs of the disease until the day when it was found recumbent in its pen. Later on it was euthanised due to lack of treatment possibilities. Pathological changes typical of avian tuberculosis were found in the liver and spleen. Mycobacterium avium ssp. avium was cultured from both organs.

The aim of study was the preliminary evaluation of the occurrence of methicillin-resistant Staphylococcus aureus (MRSA) and extended spectrum ß-lactamases (ESBL) - producing Escherichia coli in 650 milk and inflammatory secretions from cows with clinical or subclinical mastitis. One millilitre of the sample was added to Mueller-Hinton broth supplemented with 6.5% NaCl, Tryptone Soya Broth with cefoxitin and aztreonam, and then to MRSA ID agar. Presumptive MRSA colonies were analysed for the presence of mecA gene. Parallel to MRSA identification, the samples were incubated in buffered peptone water, lauryl tryptose broth and McConkey agar supplemented with cefotaxim for ESBL-producing E. coli isolation. These bacteria were identified using API Rapid 32 E and the ability of ESBL production was initially established using disc test D68C and confirmed by MIC technique using Sensititre ESBL plates. The primers (blaCTX, blaTEM, blaSHV, and blaCMY-2-group) for the detection of some of the genes encoding ESBL production were used. The 45 strains of S. aureus with mecA gene and 41 strains of E. coli with blaTEM gene were detected.

Poland is officially free of animal tuberculosis (TB), caused by Mycobacterium bovis and closely related members of the Mycobacterium tuberculosis complex, including Mycobacterium caprae. Sporadic cases of infection have been reported in cattle, but also in free-ranging species such as European bison (Bison bonasus), red deer (Cervus elaphus), Eurasian wild boar (Sus scrofa), Eurasian lynx (Lynx lynx), and Eurasian badger (Meles meles). This paper presents the history and recent findings regarding the incidence of tuberculosis in a free-living European bison herd and the possibility of interspecific transmission of the agent to other wildlife. The spoligotyping method indicated that the isolated strains belong to M. caprae showing spoligo pattern 2000037777377400. The results of these and earlier studies suggest that this M. caprae strain could be transferred from bison to wild boar.

Antimicrobial resistance was tested in Escherichia coli isolated from feces (n = 660) of red deer, roe deer, fallow deer, European bison, and wild boar shot in regional forests in Poland during two winter hunting seasons. Indicator E. coli (n = 542) was resistant against 11 of 14 tested compounds, mostly sulfamethoxazole, streptomycin, ampicillin, trimethoprim, and tetracycline (1.3-6.6% range). No significant differences were observed between boar and ruminant isolates. Most of deer and bison isolates showed no resistance. Selective screening of wildlife samples revealed 1.7% prevalence of cephalosporin-resistant E. coli found mostly in wild boars. They produced extended-spectrum beta-lactamases (bla

Introduction The aim of the study was the application and comparison of real-time PCR methods based on the fluorescence of SYBR Green I intercalating dye and TaqMan probes for the detection of the 23S rDNA gene of Listeria spp. and the hlyA gene of Listeria monocytogenes in biological samples of the liver, brain, and blood. Material and Methods Five strains of L. monocytogenes and single strains of each species L. ivanovii, L. innocua,L. grayi, L. welshimeri, and L. seeligeri were used for the experiments. Additionally, five strains of other species of bacteria were used for evaluation of the specificity of tests. In the first stage of the study SYBR Green I real-time PCRs, one allowing detection of the 23S rDNA gene and two based on the amplification the hlyA gene, were performed. In the next part, three TaqMan probe-based real-time PCRs allowing confirmation of belonging to Listeria spp. and L. monocytogenes were conducted. Results The observation of amplification curves in real-time PCRs enabled the detection of both genes. A high regression coefficient of 0.99 was found for all reactions. Specific amplification products were obtained for the 23S rDNA and hlyA genes which confirm their belonging to Listeria spp. and L. monocytogenes, respectively. Other microbial species did not reveal real-time PCR products. Conclusion Both real-time PCR methods for the detection of Listeria spp. and L. monocytogenes in biological samples demonstrated a significant sensitivity and high specificity.

The paper concerns molecular study on pathogenicity markers of fourteen Y. enterocolitica O:9 isolated from pigs in which initially positive serological reactions for brucellosis were observed (n = 41), healthy pigs, which were brucellosis-negative (n = 258), and wild boars serologically negative for brucellosis (n = 209). PCR identification proved that all isolates were ail, ystA- and myfA-positive. The plasmid encoding yadA marker was detected in nine isolates that originated from pigs serologically positive or negative for brucellosis, and from one isolate of wild boar origin. Furthermore, none of the examined isolates was ystB-positive. Results of the investigations indicate that the Y. enterocolitica O:9 isolates from pigs or wild boars, regardless of whether they were serologically positive or negative for brucellosis, may also be potentially pathogenic for humans, due to the presence of chromosomal and plasmid encoded molecular markers.

Systemic mycobacteriosis was diagnosed in a group of ornamental fish. Although a large number of acid-fast bacterial rods were identified in the kidneys, liver, and muscles of each fish, no granulomas were observed in internal organs. Mycobacterium peregrinum was identified using the GenoType Mycobacterium CM assay. This study illustrates a considerable risk of atypical mycobacteriosis in humans.

Fluorescence polarisation assay (FPA) was evaluated as a potential tool improving specificity of serological diagnosis of brucellosis in cattle and pigs. The evaluation was performed by comparing the results of FPA with the results of rose Bengal test (RBT), serum agglutination test (SAT), complement fixation test, and indirect ELISA when problematic sera, regarded as false positive, were tested. Four hundred and seventy-five sera, including 201 porcine and 274 bovine samples, reacting positively in at least one classical serological assay were used. Only six bovine sera were FPA positive (two positive in SAT and RBT and four positive in SAT only). Different situation was observed when porcine sera were examined. Out of 201 sera, 109 were also positive in FPA. The studies confirmed that in cattle FPA enables to reduce highly the number of false positive reactions for brucellosis. On the other hand, in pigs, the method is a little more specific in comparison to other methods applied.

Bovine tuberculosis is an infectious disease that occurs in many species of both domestic and wild animals, as well as those held in captivity. The etiological factor is the acid resistant bacillus (Mycobacterium bovis or Mycobacterium caprae), which is characterized by the major pathogenicity among mycobacteria belonging to the Mycobacterium tuberculosis complex. The material from 8 antelopes from the zoo, suspected for tuberculosis were examined, and M. bovis strains were isolated from 6 of them. The spoligotyping method showing spoligo pattern 676763777777600. In Poland, this spoligotype has not been observed so far.

The paper presents the results of bacteriological and molecular investigations on the presence of Y. enterocolitica O:9 in the head, mammary and genital lymph nodes, spleen, liver, and uterus samples originating from 58 cows slaughtered due to the positive results of serological examinations for brucellosis. All samples were cultured for Brucella and Y. enterocolitica and examined in multiplex PCR assay (mPCR), in order to identify the universal 16S rRNA Brucella sp. marker and amplify the perosamine synthetase (per) gene, specific for Y. enterocolitica O:9 only. Out of 58 examined animals, in 23 cases the presence of Y. enterocolitica was demonstrated. Typical Yersinia-suspected colonies were seen after 24-48 h incubation on Cefsulodin-Irgasan- Novobiocin (CIN) Yersinia selective solid medium. The mPCR analysis confirmed the presence of predicted amplicon of 312 bp, typical for Y. enterocolitica O:9 in 20 out of 58 lymph node samples tested and similarly, as in bacteriological examination, other samples were negative. The presence of the 16S rRNA gene of Brucella, generating the amplicon of 905 bp, was not observed in any of the samples tested.

The aim of the study was to perform a molecular investigations for the presence of pathogenicity genotypic markers of Y. enterocolitica O:9 isolated from cattle, in which initially positive serological reactions for brucellosis were observed. Almost all isolates were ail-, ystA- and myfA-positive (n=19). On the other hand, one isolate, which harboured plasmid encoding gene yadA was ail- , ystA- and myfA-negative. The plasmid encoding yadA marker was present in half of the isolates tested. None of the examined isolates was ystB-positive. The results of the investigations revealed that the Y. enterocolitica O:9 isolates, related to false positive serological results for brucellosis, may be also potentially pathogenic for humans, due to the presence of chromosomal and plasmidencoded molecular markers.

The aim of this study was the evaluation of fluorescence polarisation assay (FPA) in the diagnosis of porcine brucellosis in comparison with Rose Bengal test (RBT), serum agglutination test (SAT), complement fixation test (CFT), 2-mercaptoethanol test, and ELISA. Eight hundred seventeen sera from pigs, including 612 sera from healthy animals, seven sera from Brucella suis bv2 culture positive animals, and 198 sera classified as false positive, originated from confirmatory investigations, were used. All sera from healthy animals, negative in RBT, SAT, CFT, and ELISA were also negative in FPA. All sera positive in serological examination, originated from Brucella infected animals, were also positive in FPA. Among sera classified as false positive almost half of the samples tested (49.49%) reacted positively in FPA. The examinations confirmed the usefulness of FPA in diagnosis of porcine brucellosis, but the method, like the other tests, does not allow resolving the problem of discrimination cross-reacting from specific antibodies.

Bovine tuberculosis (bovine TB, bTB) is caused by bovine bacilli: Mycobacterium bovis and M caprae The studies conducted in Poland, in the National Bovine Tuberculosis Reference Laboratory in the Department of Microbiology of the National Veterinary Research Institute in Pulawy, show that animal tuberculosis in Poland is also caused by M caprae We here describe the identification and genotypic assessment of 52 isolates of M caprae obtained from Polish cattle and wild animals over the last five years. We show that strains isolated from bison have significant genotypic diversity and are distinct compared with the genotypes of strains isolated from cattle. Similarly, isolates from cattle herds can be highly genotypically variable. Formal designation of the members of the Mycobacterium tuberculosis complex is controversial in Poland; there is a gap in veterinary legislation with regard to bTB and no explicit mention of M caprae causing tuberculosis in animal.

Since 2009, Poland has had a TB-free status, although over the last seven years 12-34 cases of bovine TB have been recorded annually. In 2009-2012 the largest number of cattle infected with Mycobacterium bovis and Mycobacterium caprae were culled in Masovian Voivodeship. Likewise, the largest number of sources of this zoonosis were recorded in that voivodeship. The vicinity of farms where bTB was found indicated that it could have been transmitted between their herds. The aim of this study was to characterise the molecular patterns of bovine bacillus strains isolated from cattle in Masovian Voivodeship and the molecular relationships between them. The material for microbiological examination came from 38 cattle (Bos taurus) located in 7 counties of Masovian Voivodeship. These 38 strains of MTBC were further identified as M. bovis (24 isolates; 63%) and M. caprae (14 isolates; 37%). A two-step genotyping analysis of the 38 MTBC strains identified 24 molecular patterns, closely related phylogenetically, which were assigned to 8 clusters of 2-6 strains. Sources of transmission were identified in 8 out of 13 herds examined in the 7 counties of Masovian Voivodeship. The results of the genotyping analysis excluded the possibility of TB transmission between different herds in Masovian Voivodeship. It was proved, however, that TB had been transmitted between animals bred on one of the farms..

Resistance to β-lactams is considered one of the major global problems and recently it became the most frequently studied topic in the area of antimicrobial resistance. The study was focused on phenotypic and genetic characterisation of commensal Escherichia coli (E. coli), including those producing cephalosporinases, isolated from gut flora of healthy slaughter animals. E. coli were cultured simultaneously on MacConkey agar (MCA) and cefotaxime supplemented MCA. The isolates were confirmed with ONPG and indol tube tests as well as PCR targeting uspA gene. Microbroth dilution method was applied for determination of Minimal Inhibitory Concentrations and interpreted according to EUCAST epidemiological cut-off values. Cephalosporin resistance phenotypes were defined by E-tests (BioMerieux) and relevant gene amplicons from selected strains were sequenced. A total of 298 E. coli isolates with cephalosporin resistance (ESC) found in 99 ones, were obtained from 318 cloacal or rectal swabs deriving from broilers, layers, turkeys, pigs and cattle. Both extended spectrum β-lactamase (ESBL) and ampC-cephalosporinase resistance phenotypes were noted in all tested animal species but cattle. At least one of the analysed genes was identified in 90 out of 99 cephalosporin-resistant isolates: blaTEM (n=44), blaCMY (n=38), blaCTX-M (n=33) and blaSHV (n=12). None of the phenotypes was identified in nine isolates. Sequencing of PCR products showed occurrence of ESBL-genes: blaCTX-M-1/-61, blaSHV-12, blaTEM-1,-52/-92,-135 and ampC-gene blaCMY-2. They were located on numerous and diverse plasmids and resistance transferability was proved by electroporation of blaSHV-12 and blaCTX-M-1/-61 located on X1 plasmids. Detection of cephalosporin resistant E. coli confirms the existence of resistance genes reservoir in farm animals and their possible spread (i.e. via IncX1 plasmids) to other bacteria including human and animal pathogens. The identified genetic background indicates on ecological aspects of selection and dissemination of cephalosporin resistance in E. coli isolated from food-producing animals rather than its potential role for public health threats.

Poland is officially tuberculosis free and bovine tuberculosis (BTB) cases are rarely found except in bovids. We found BTB in a wild boar (Sus scrofa) in the Bieszczady Mountains, southeastern Poland. Studies suggest possible transmission of infection between free-living European bison (Bison bonasus caucasicus) and wild boar in this area.

Bovine brucellosis is an infectious disease caused by bacteria of the Brucella genus, primarily by B. abortus, less frequently by B. melitensis, and occasionally by B. suis. In the European Union, brucellosis in cattle has been eradicated in most of the Member States, which are recognized as 'officially free from bovine brucellosis'. Nevertheless, cattle herds continue to be serologically monitored for the potential re-emergence of the disease. The aim of the presented study was to show the results of bacteriological investigations of cattle slaughtered in Poland in years 2002-2011 on account of positive serological reactions for brucellosis. Specimens (sera and tissues) from 176 cows were examined. Sera from the animals were tested using RBT(rose bengal test), SAT (serum agglutination test), CFT (complement fixation test), 2-ME (2-mercaptoethanol test), Coombs (Coombs antiglobulin test) and ELISA (enzyme linked immunosorbant assay). Tissue samples were cultured for Brucella, according to official protocols. All sera were RBT and SAT-positive, 170 of them were CFT- positive, whereas 6 other samples were CFT negative while positive in Coombs and ELISA. In bacteriological examination, B. abortus was not isolated. On the other hand, B. suis biovar 2 was isolated from 5 cows, which had never been reported previously in Poland. Three cows came from the same herd. Conventional, as well as, molecular investigations based on PCR methods, confirmed that the bacteria isolated bacteria belong to the B. suis biovar 2. In Poland, as in many other European countries, wildlife (wild boars and hares) constitutes a huge reservoir of the said biovar. The results of the presented research indicate that B. suis biovar 2 can easily infect cattle, and undoubtedly plays a role in the epidemiology and control of bovine brucellosis.

Forty four pigs with typical characteristics for false positive serological reactions (FPSR) were examined for the presence of Yersinia enterocolitica O:9. The positive reactions were observed in rose bengal test (RBT, N=23 sera), serum agglutination test (SAT, N=16), complement fixation test (CFT, N=9), indirect ELISA (i-ELISA, N=11) in first, and in RBT (N=14), SAT (N=8), CFT (N=7) and i-ELISA (N=18) in second examination, respectively. In bacteriological examination Y. enterocolitica was confirmed in 12 cases. Six of these isolates were identified with PCR as Y. enterocolitica O:9.

Monitoring of antimicrobial resistance in commensal Escherichia coli (N = 3430) isolated from slaughtered broilers, laying hens, turkeys, swine, and cattle in Poland has been run between 2009 and 2012. Based on minimal inhibitory concentration (MIC) microbiological resistance to each of 14 tested antimicrobials was found reaching the highest values for tetracycline (43.3%), ampicillin (42.3%), and ciprofloxacin (39.0%) whereas the lowest for colistin (0.9%), cephalosporins (3.6 ÷ 3.8%), and florfenicol (3.8%). The highest prevalence of resistance was noted in broiler and turkey isolates, whereas it was rare in cattle. That finding along with resistance patterns specific to isolation source might reflect antimicrobial consumption, usage preferences or management practices in specific animals. Regression analysis has identified changes in prevalence of microbiological resistance and shifts of MIC values. Critically important fluoroquinolone resistance was worrisome in poultry isolates, but did not change over the study period. The difference (4.7%) between resistance to ciprofloxacin and nalidixic acid indicated the scale of plasmid-mediated quinolone resistance. Cephalosporin resistance were found in less than 3.8% of the isolates but an increasing trends were observed in poultry and MIC shift in the ones from cattle. Gentamycin resistance was also increasing in E. coli of turkey and cattle origin although prevalence of streptomycin resistance in laying hens decreased considerably. Simultaneously, decreasing MIC for phenicols observed in cattle and layers isolates as well as tetracycline values in E. coli from laying hens prove that antimicrobial resistance is multivariable phenomenon not only directly related to antimicrobial usage. Further studies should elucidate the scope of commensal E. coli as reservoirs of resistance genes, their spread and possible threats for human and animal health.

The aim of the study was an evaluation of fluorescence polarisation assay (FPA) as a potential tool improving specificity of serological diagnosis of brucellosis in pigs. The evaluation was done by comparing the results of FPA with the results of rose bengal test (RBT), serum agglutination test (SAT), complement fixation test (CFT) and ELISA when false positive sera were tested. One hundred ninety porcine samples, reacting positively in at least one classical serological assay were used. We observed that among 198 sera, 104 were also positive in FPA. The studies confirmed that porcine FPA adds little as far as specificity in comparison to other methods is concerned.

The purpose of the study was to define genetic diversity of reptilian Salmonella enterica serovar (S.) Kentucky isolates and their epidemiological relations to the ones from poultry, food, and environmental origin in Poland. Between 2010 and 2012 twenty-four S. Kentucky isolates derived from snakes (N = 8), geckos (N = 7), chameleons (N = 4), agamas (N = 1), lizard (N = 1), and environmental swabs taken from reptile exhibition (N = 3) were identified. They were characterized with antimicrobial minimal inhibitory concentration testing, XbaI–PFGE and MLST typing. The profiles compared to S. Kentucky available in BioNumerics local laboratory database (N = 40) showed 67.3% of relatedness among reptile isolates. Three genetic lineages were defined. The first lineage gathered 20 reptile isolates with 83.4% of similarity and wild-type MICs for all antimicrobials tested but streptomycin in single case. The remaining three reptilian and one post-exhibition environment S. Kentucky isolates were clustered (87.2%) with isolates originating from poultry, mainly turkey, food, and environment and presented variable non-wild type MICs to numerous antimicrobials. The third S. Kentucky lineage was composed of two isolates from feed (96.3%). The results suggest diverse sources and independent routes of infection. Most of the isolates belonged to reptile-associated clones spread both horizontally and vertically. Simultaneously, PFGE profiles and MLST type indistinguishable from the ones observed in poultry point out carnivore reptiles as possible vector of infection with multidrug and high-level ciprofloxacin resistant (MIC ≥ 8 mg/L) S. Kentucky. Public awareness and education are required to prevent potential reptile-associated S. Kentucky infections in humans.

A total of 42 Brucella strains were isolated from animals in Poland in years 2003-2012. Most of them (N=37) originated from wild animals, 3 from cattle, 1 from pig and 1 from sheep. The strains were characterised using both bacteriological and molecular (Bruce-ladder and MLVA) methods. The examinations revealed that all strains from wild boars, hares, cattle and pigs (N=41) had the same phenotypic characteristics and were classified as B. suis biovar 2. The remaining strain, isolated from sheep, was classified as B. ovis. The molecular examination showed that all B. suis biovar 2 strains, except one, had the same molecular profile as reference strain B. suis bv2 Thomsen. Different from the others strain originated from boars imported to Poland and its VNTR profile was typical for Iberian strains.

Free-living snakes may carry a broad range of pathogens that differ considerably by geographical locations and source species. The biological role of wildlife snakes for transmission of bacteria and helminths is hardly explored. The purpose of the study was a snapshot on the zoonotic agents load in free-living snakes found dead in a landscape park in central Poland. A total of 16 dead free-living snakes identified as European grass snake (Natrix natrix, N = 15) and a smooth snake (Coronella austriaca, N = 1) were tested. Abdominal organs were used for bacteriological testing, whereas, parasites were searched for in the remaining carcasses. Fourteen (87.5 %) individuals were positive for Salmonella spp., whereas, trematodes were found in 13 out of 16 samples (81.3 %). A total of 33 isolates representing 11 Salmonella serovars or antigenic forms were observed with Salmonella enterica subsp. diarizonae (IIIb) 38:r:z being the most frequent. The trematodes were recognized as Alaria alata mesocercariae (Distomum musculorum suis, DMS). Additionally, unidentified nematodes were detected in four samples. No Yersinia spp. and Mycobacterium spp. were found. The results indicate huge burden of Salmonella spp. and muscle parasites carriage by snakes present in the natural environment and indicate possible zoonotic and epidemiological impact. It justifies our concept of free-living snakes as possible source and vector of pathogens, especially for omnivores and scavengers foraging on snake corpses. Finding of rare and diverse Salmonella serovars and unidentified nematodes proves the need for in-depth studies and systematic approach to reveal the role of free-living reptiles in epidemiology of infectious agents.