Seroprevalence of Selected Zoonotic Agents among Hunters from Eastern Poland.

The aim of our study was the collection of seroprevalence data for Toxoplasma gondii, Coxiella burnetii, Trichinella spp., and Francisella tularensis from hunters in Lublin Province. The antibodies against T. gondii and C. burnetii were recorded in 38.5% and 16.2% of the sera, respectively. 4.05% of the sera were seropositive for both T. gondii and C. burnetii. None of the sera tested reacted positively with F. tulariensis or Trichinella spp. Seroprevalence of T. gondii and C. burnetii is common among the hunters from Lublin Province. It seems reasonable to undertake similar research among hunters from other regions of eastern Poland.

The Serological Surveillance of Hepatitis E virus among Hunters and Foresters in Eastern Poland.

The aim of the study was the analysis of the occurrence of specific antibodies against HEV among hunters and foresters who are at risk to be exposed. The study group consisted of 210 hunters (23-80 years old) and 60 foresters (22-64 years old). Anti-HEV IgG were present in 3.81% of the samples of the hunters and in 5% of the samples of the foresters. The statistical analysis shows no significant differences in the results anti-HEV IgG between the groups of hunters and foresters (p = 0.5278). Significantly higher anti-HEV IgG titers were found in the older age group (> 55 years old).

Tuberculosis in Antelopes in a Zoo in Poland--Problem of Public Health.

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.

[Candida species at the workplace: microbiota component, opportunistic pathogen and zoonotic agent]. / Candida species w miejscu pracy: element mikrobioty, patogen oportunistyczny oraz czynnik zoonotyczny.

Candida spp. isolated from both humans and animals have a similar genotype. Properties of Candida spp. specific for different host species have not been isolated, followed by studies indicating that animals can be a reservoir of these fungi for humans. Occupational exposure concerns workers who have direct contact with farm animals, i.e., farmers, breeders, veterinarians, farm technicians. Hand dermatitis and fungal infection may be caused by prolonged exposure to water and occlusive gloves. The risk of fungal infection is estimated to be high for seafood workers, florists, hairdressers, bakers and cooks, gastronomy workers and healthcare workers. Even though Candida spp. are effective as saprophytic, in the event of a weakening of the function or disturbance of homeostasis, the risk of developing an additional form of candidiasis is increasing due to the intensification of animal production, environmental changes and the excessive use of antibiotics to treat infections in humans and animals. Employers and workers should adopt appropriate strategies to reduce factors conductive to Candida spp. infection at professional work. Med Pr Work Health Saf. 2023;74(5):425-33.

Mycobacterial Infections in Invasive Turtle Species in Poland.

Over the last 30 years, the number of invasive turtle species living in the wild has significantly increased in Poland. This proliferation carries many threats, which mainly include the displacement of native species of animals from their natural habitats. Turtles can also be reservoirs for pathogens, including bacteria from the Mycobacterium genus. In order to confirm or rule out the presence of acid-fast mycobacteria in the population of invasive turtle species, samples from carapace, plastron, internal organs and mouth cavity swabs from 125 animals were tested. Twenty-eight mycobacterial strains were isolated in culture, which were classified as atypical following multiplex-PCR reactions. The GenoType Mycobacterium Common Mycobacteria (CM) test, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, PCR-restriction fragment length polymorphism (PRA)-hsp65 and DNA sequencing were used to identify the species of isolates. Of the 28 strains, 11 were identified as M. fortuitum, 10 as M. chelonae, 3 as M. avium ssp. avium, 2 as M. nonchromogenicum and 1 each of M. neoaurum and M. scrofulaceum. The results of the research will also strengthen the understanding that these animals can be vectors for pathogens when living in the wild.

The Eradication of M. Caprae Tuberculosis in Wild Boar (Sus Scrofa) in the Bieszczady Mountains, Southern Poland - An Administrative Perspective.

Introduction: Animal tuberculosis (TB) is a zoonotic disease caused by acid-fast bacteria belonging to the Mycobacterium tuberculosis complex (MTBC). Both animals and humans are susceptible to infection by the MTBC. Interspecies transmission is also possible, including to livestock and humans. In the years 1997-2013, many tuberculosis cases were recorded in European bison in the Bieszczady Mountains; more alarmingly, TB was also recorded in wild boar in the years 2013-2020. Material and Methods: In the years 2013-2020, 104 wild boar from the Bieszczady Mountains were tested for TB through necropsy, mycobacterial culture, strain identification and spoligotyping. Results: The microbiological examination confirmed TB in 46 wild boar; these infections were identified as M. caprae, spoligotype SB2391. Conclusion: Free-living European bison are at risk of TB infection from wild boar carrying M. caprae. This situation also poses a risk to local cattle. There is a need for further activities aimed at monitoring the disease, preventing further transmission, and minimising the risk to public health.

Ten Years of Animal Tuberculosis Monitoring in Free-Living European Bison (Bison bonasus) in Poland.

In the period 1996-2012, two outbreaks of animal tuberculosis were noted in the population of free-living European bison (Bison bonasus caucasicus) in the Bieszczady Mountains, Southern Poland. As the European bison is an endangered species and particularly susceptible to tuberculosis, not to mention a national icon, the decision was made to test all deceased bison for TB in Poland. The screened bison were obtained by elimination due to poor health or natural death. A total of 159 European bison have been examined over the last 10 years. The individuals came from four regions of Poland (Bialowieza Forest, Bieszczady Mountains, Borecka Forest, Knyszynska Forest), not only from the area where tuberculosis is still endemic. Mycobacterium bovis and Mycobacterium avium spp. hominisuis were identified in two different herds. The isolation of M. bovis from European bison was the first case described in Poland. So far, the only causative agent of tuberculosis identified in European bison in Poland, both in the wild and in captive herds, was Mycobacterium caprae. The isolated M. bovis spoligotype has not previously been registered in international spoligotype databases so far. The obtained results highlight the need to monitor TB in European bison in Poland.

The Potential Risk of International Spread of Mycobacterium Bovis Associated with Movement of Alpacas.

Introduction: The study highlights the transboundary nature of tuberculosis (TB) in alpacas and the failure of current ante-mortem testing protocols (the tuberculin skin and Enferplex Camelid TB tests) to identify TB-free alpaca herds and individuals for export. Our research and the available literature indicate that the alpaca (Vicugna pacos) is extremely susceptible to Mycobacterium bovis infection, and that testing periodicity fails to take into account that animals do not manifest disease symptoms for a long time. The skin test failed to identify Mycobacterium bovis infection in two alpacas prior to their movement from the UK to Poland. The animals were purchased by a breeding centre in Poland, and were then shown at an international animal exhibition. The last owner of the alpacas before their deaths from TB bought the infected animals unwittingly in order to run rehabilitation activities with disabled children on his farm. Material and Methods: Thoracic lymph node, lung and liver tissue samples obtained at necropsy were examined histopathologically after Ziehl-Neelsen staining. Tissue samples were homogenised and mycobacteria present there were cultured on Stonebrink's medium during a 6-week incubation. A commercial test using polymorphism of the chromosomal direct repeat region provided species identification and additional identification was by spacer oligonucleotide typing and mycobacteria interspersed repetitive unit-variable number tandem repeat analysis with a gel electrophoresis protocol. Results: The microbiological examination confirmed multiorgan TB caused by the SB0666 spoligotype of Mycobacterium bovis. Conclusion: Due to the suboptimal performance of current diagnostic tests for TB in alpacas, there is a risk that infected animals may be moved unwittingly. A risk of TB spread associated with the international movement of alpacas is implied by this study.

Mycobacterium bovis Transmission between Cattle and a Farmer in Central Poland.

INTRODUCTION: Zoonoses have recently become an increasing public health problem. Zoonoses are estimated to account for 60% of all emerging infectious diseases. One particularly important zoonosis is human tuberculosis, especially tuberculosis due to Mycobacterium bovis (M. bovis), which is naturally resistant to pyrazinamide (PZA). MATERIAL AND METHODS: The patient had a pulmonary form of tuberculosis accompanied by a cough and fever. At the same time, the disease was also confirmed in 20 out of 25 cattle on the farm. The clinical specimen (sputum) was examined in accordance with the European Union (EU) laboratories' methodology. Tissue materials from cattle were verified in the National Veterinary Research Institute (NVRI), in the Bovine tuberculosis (BTB) Reference Laboratory, Pulawy, Poland and tested in accordance with the guidelines for the laboratory diagnosis of BTB. RESULTS: All M. bovis isolates represented one spoligotype, SB0120. The results of mycobacterial interspersed repetitive unit variable number tandem repeat (MIRU-VNTR) evaluation showed the same genetic pattern. CONCLUSIONS: Findings from this study suggest the first confirmed interspecific transmission of Mycobacterium bovis, between a farmer and his cattle, in Poland. Present findings support the increasing concern regarding zoonotic TB that has been highlighted elsewhere.

Adaptation of Brucella melitensis Antimicrobial Susceptibility Testing to the ISO 20776 Standard and Validation of the Method.

Brucellosis, mainly caused by Brucella (B.) melitensis, is associated with a risk of chronification and relapses. Antimicrobial susceptibility testing (AST) standards for B. melitensis are not available, and the agent is not yet listed in the EUCAST breakpoint tables. CLSI recommendations for B. melitensis exist, but they do not fulfill the requirements of the ISO 20776 standard regarding the culture medium and the incubation conditions. Under the third EU Health Programme, laboratories specializing in the diagnostics of highly pathogenic bacteria in their respective countries formed a working group within a Joint Action aiming to develop a suitable method for the AST of B. melitensis. Under the supervision of EUCAST representatives, this working group adapted the CLSI M45 document to the ISO 20776 standard after testing and validation. These adaptations included the comparison of various culture media, culture conditions and AST methods. A Standard Operation Procedure was derived and an interlaboratory validation was performed in order to evaluate the method. The results showed pros and cons for both of the two methods but also indicate that it is not necessary to abandon Mueller-Hinton without additives for the AST of B. melitensis.

[The evaluation of hunters and foresters' knowledge of the possible ways of preventing Borrelia burgdorferi infections]. / Ocena wiedzy mysliwych i lesników w zakresie mozliwosci zapobiegania infekcjom Borrelia burgdorferi.

BACKGROUND: This study is aimed at evaluating the awareness among hunters and foresters concerning the activities undertaken for the prevention of Borrelia burgdorferi infections. MATERIAL AND METHODS: The study group included 236 hunters from Poland and 339 foresters from Ukraine. The study was based on an original questionnaire including questions which concerned the applied methods of removing ticks attached to the skin, the kind of preventive measures undertaken to avoid tick bites, the evaluation of knowledge concerning the prevention of tick-borne diseases, as well as the participants' willingness to be educated on this issue. RESULTS: Removing a tick with a pair of tweezers by a simple vertical move was more frequently practiced by the hunters (16.5%). Spreading an oily substance over a tick to make it go out on its own was more often applied by the foresters (18.8%). The occurrence of erythema migrans was more frequently reported by the foresters (47.2%). Among the persons declaring no knowledge of the prevention of tick-borne diseases, 77.8% of the hunters and 61.5% of the foresters were willing to gain such knowledge, and 11.1% of the hunters and 21.2% of the foresters did not want any education on this issue. The hunters more often than the foresters applied repellents and checked their body after returning from green areas. CONCLUSIONS: Education is needed in certain groups undertaking hobby or vocational activities in forest areas of Poland and Ukraine. It seems reasonable to undertake research on the preferences concerning the choice of repellents among the groups at risk of tick bites. Med Pr. 2020;71(1):59-68.

Molecular Characterisation of the Mycobacterium Bovis Causing Bovine Tuberculosis Outbreaks in Poland.

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.

Occurrence of Enterococci in Mastitic Cow's Milk and their Antimicrobial Resistance.

INTRODUCTION: The aim of the study was to evaluate the occurrence of enterococci in inflammatory secretions from mastitic bovine udders and to assess their antimicrobial resistance. MATERIAL AND METHODS: A total of 2,000 mastitic milk samples from cows were tested in 2014-2017. The isolation of enterococci was performed by precultivation in buffered peptone water, selective multiplication in a broth with sodium azide and cristal violet, and cultivation on Slanetz and Bartley agar. The identification of enterococci was carried out using Api rapid ID 32 strep kits. The antimicrobial susceptibility was evaluated using the MIC technique. RESULTS: Enterococci were isolated from 426 samples (21.3%). Enterococcus faecalis was the predominant species (360 strains), followed by E. faecium (35 isolates), and small numbers of others. The highest level of resistance was observed to lincomycin, tetracycline, quinupristin/dalfopristin (Synercid), erythromycin, kanamycin, streptomycin, chloramphenicol, and tylosin. Single strains were resistant to vancomycin and ciprofloxacin. All isolates were sensitive to daptomycin. E. faecalis presented a higher level of resistance in comparison to E. faecium, except to nitrofurantoin. CONCLUSION: The results showed frequent occurrence of enterococci in mastitic cow's milk and confirmed the high rate of their antimicrobial resistance.

Human as a potential vector of bovine tuberculosis in cattle.

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.

Nontuberculous mycobacterial skin disease in cat; diagnosis and treatment - Case report.

INTRODUCTION: Mycobacterial diseases of humans and animals can be caused by mycobacteria other than tuberculosis (MOTT). The transmission of the infection primarily occurs via the respiratory or oral routes, but also via a damaged skin barrier. MOTT have high resistance to external factors; therefore, infected, undiagnosed animals can pose a risk for public health. CASE REPORT: The case study describes mycobacterial skin infection in a domestic cat. The correct diagnosis was reached four months after the appearance of the first clinical signs. Those were purulent, granulomatous lesions and fistulas, which could potentially act as a source of the infection for the owners and the veterinarian who cared for the animal. CONCLUSION: Despite using advanced diagnostic techniques, establishing the final cause of the cat's illness was a lengthy process. The skin lesions could contribute to the transmission of the bacteria in the environment. Non-targeted treatments could also cause antimicrobial resistance.

Treatment for active tuberculosis in giraffe (Giraffa camelopardalis) in a Zoo and potential consequences for public health - Case report.

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.

Molecular characterisation of Mycobacterium caprae strains isolated in Poland.

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.

Prevalence of Borrelia Burgdorferi Sensu Lato in Ticks from the Ternopil Region in Ukraine.

INTRODUCTION: Lyme borreliosis/Lyme disease is caused by Borrelia burgdorferi and is one of the most common vector-borne diseases transmitted by ticks. MATERIAL AND METHODS: A total of 136 Ixodes ricinus ticks, collected in the Ternopil (Ukraine) region, including 126 adults (70 females and 56 males), and 10 nymphs were examined. The identification of the species and their developmental form was based on morphological characteristics. RESULTS: PCR with B5S-Bor and 23S-Bor primers resulted in Borrelia burgdorferi sensu lato DNA amplification among six ticks (4.4%). The detailed analysis based on the DNA sequencing showed the presence of DNA of Borrelia afzelii in four samples; the remaining two represented Borrelia burgdorferi sensu lato complex, although their genospecies were not determined. The research confirmed the dominance of Borrelia afzelii genospecies in the ticks from Ukraine. CONCLUSION: It seems reasonable to undertake similar research in ticks from other regions of Ukraine. Knowledge in this field can be useful for public health and planning the prevention of tick-borne diseases.

Evaluation of Susceptibility to Antimycobacterial Drugs in Mycobacterium Tuberculosis Complex Strains Isolated from Cattle in Poland.

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.

Development of a multiplex PCR (m-PCR) test for rapid identification of genes encoding heat-labile (LTI) and heat-stable (STI and STII) toxins of enterotoxigenic Escherichia coli (ETEC) with internal control of amplification.

A multiplex PCR system was developed for specific identification of genes encoding heat-labile (LTI) and heat-stable (STI and STII) toxins of enterotoxigenic Escherichia coli (ETEC) strains. In addition, primers specific for the E. coli gene coding for 16S rRNA were used as an internal control of the DNA amplification. The specificity of the method was validated by single PCR tests performed with reference to E. coli strains as well as pig-isolated bacteria and 100% correlation was observed. The developed multiplex PCR allowed rapid and specific identification of enterotoxin-positive E. coli and may be used as a sensitive and specific method for a direct determination of ETEC and to differentiate them from other E. coli isolates.