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Introduction Bovine tuberculosis is one of the most dangerous zoonotic diseases. Despite the near-complete elimination of the disease from cattle breeding in Poland achieved in 2009, its re-emergence is now observed. Globally, the number of human cases is underestimated and the importance of free-living animals as reservoirs of tuberculosis is growing. As a species highly susceptible to Mycobacterium tuberculosis complex infection, the European bison (Bison bonasus) has a role in the transmission of the disease in Poland. The purpose of the investigation was to assess the epidemiological situation of tuberculosis in Polish European bison serologically. Material and Methods A total of 460 serum samples were collected from 436 European bison from 15 out of 26 national populations between 2013 and 2020. An M. bovis ELISA was used, and its sensitivity and specificity were assessed with an eyelid tuberculin skin test (TST) and interferon gamma release assay (IGRA). Results Mycobacterium bovis antibodies were detected in nine serum samples. The presence of antibodies was found in two animals from the Białowieża Forest (1.2% of the population), and one each from the Borecka Forest (2.4%) and the Warsaw Zoo (14.3%). One European bison among the 14 sampled (7.1%) from Smardzewice was positive on five occasions. Other samples from Smardzewice and the Bieszczady Mountains, where tuberculosis had previously been reported, were negative. Conclusion ELISA testing is an effective, easy and cost-efficient tool for monitoring of tuberculosis-naïve populations. Serological testing in tuberculosis control programmes can significantly improve the detection of infected herds. Antibody ELISAs may supplement TST and IGRA, but cannot replace them.

The spread of antibiotic-resistant pathogens and the resurgence of tuberculosis disease are major motivations to search for novel antimicrobial agents. Some promising candidates in this respect are cationic polymers, also known as synthetic mimics of antimicrobial peptides (SMAMPs), which act through the membrane-lytic mechanism. Development of resistance toward SMAMPs is less likely than toward currently employed antibiotics; however, further studies are needed to better understand their structure-activity relationship. The main objective of this work is to understand the cross-influence of hydrophobicity, main-chain flexibility, and the topology of ionenes (polycations containing a cationic moiety within the main-chain) on activity. To fulfill this goal, a library of ionenes was developed and compared with previously investigated molecules. The obtained compounds display promising activity against the model microorganisms and drug-resistance clinical isolates, including

Background Tuberculosis (TB) is a highly prevalent disease, with millions of people dying yearly. The fight against TB requires surveillance of the population of strains circulating worldwide and the analysis of the prevalence of certain strains in populations. Nowadays, whole genome sequencing (WGS) allows for accurate tracking of TB transmission. Currently, there is a lack of a comprehensive summary of the characteristics of TB outbreaks. Methods We systematically analyzed studies reporting TB outbreaks worldwide, monitored through WGS of Mycobacterium tuberculosis. We 1) mapped the reported outbreaks from 2011–2020, 2) estimated the average size of the outbreaks, 3) indicated genetic lineages causing the outbreaks, and 4) determined drug-resistance patterns of M. tuberculosis strains involved in the outbreaks. Results Most data originated from Europe, Asia, and North America. We found that TB outbreaks were reported throughout the globe, on all continents, and in countries with both high and low incidences. The detected outbreaks contained a median of five M. tuberculosis isolates. Most strains causing the outbreaks belonged to lineage four, more rarely to lineage two. Reported outbreak isolates were often drug resistant. Conclusions We conclude that more WGS surveillance of M. tuberculosis outbreaks is needed. Globally standardized procedures might improve the control of M. tuberculosis infections.

Mycobacterium chimaera is the newly described species belonging to Mycobacterium avium complex (MAC), with morphology and growth characteristics closely related to Mycobacterium intracellulare. The aim of this retrospective study was to analyze the frequency and clinical significance of M. chimaera identification in the population of patients with previous positive respiratory cultures for M. intracellulare or MAC. 200 strains of M. intracellulare or MAC, isolated from respiratory specimens of patients hospitalized in pulmonary wards, between 2011 and 2020, were retrospectively analyzed with GenoType NTM-DR test. 88 (44%) of strains were re-classified to M. chimaera species. Analysis of clinical data in 30 patients with positive M. chimaera isolates revealed that they were diagnosed with chronic obstructive pulmonary disease (COPD) – 27%, past tuberculosis – 20%, or interstitial lung diseases – 17%, respectively. Non-tuberculous mycobacterial lung disease (NTMLD) caused by M. chimaera has been recognized in 53% of patients, most often in those presenting with post-tuberculous lung lesions. M. chimaera was almost exclusively isolated from respiratory specimens of patients with underlying lung diseases, especially those with COPD and/or past tuberculosis. NTMLD due to M. chimaera was diagnosed predominantly in patients with past tuberculosis.

Rapidly growing mycobacteria (RGM) cause an increasing international concern, mainly due to their natural resistance to many antibiotics. The aim of this study was to conduct species identification and determine the antimicrobial susceptibility profiles of RGM isolated in Poland. Antimicrobial susceptibility was tested using broth microdilution and the RAPMYCOI panel. A total of 60 strains were analysed, including the following species

Novel conjugates (CP) of moxifloxacin (MXF) with fatty acids (1m–16m) were synthesized with good yields utilizing amides chemistry. They exhibit a more pronounced cytotoxic potential than the parent drug. They were the most effective for prostate cancer cells with an IC50 below 5 µM for respective conjugates with sorbic (2m), oleic (4m), 6-heptenoic (10m), linoleic (11m), caprylic (15m), and stearic (16m) acids. All derivatives were evaluated against a panel of standard and clinical bacterial strains, as well as towards mycobacteria. The highest activity towards standard isolates was observed for the acetic acid derivative 14m, followed by conjugates of unsaturated crotonic (1m) and sorbic (2m) acids. The activity of conjugates tested against an expanded panel of clinical coagulase-negative staphylococci showed that the compound (14m) was recognized as a leading structure with an MIC of 0.5 μg/mL denoted for all quinolone-susceptible isolates. In the group of CP derivatives, sorbic (2) and geranic (3) acid amides exhibited the highest bactericidal potential against clinical strains. The M. tuberculosis Spec. 210 strain was the most sensitive to sorbic (2m) conjugate and to conjugates with medium- and long-chain polyunsaturated acids. To establish the mechanism of antibacterial action, selected CP and MXF conjugates were examined in both topoisomerase IV decatenation assay and the DNA gyrase supercoiling assay, followed by suitable molecular docking studies.

Background The majority of animal tuberculosis (TB) cases reported in wildlife in Poland over the past 20 years have concerned the European bison inhabiting the Bieszczady Mountains in Southeast Poland: an area running along the border of Southeast Poland. As no TB cases have been reported in domestic animals in this region since 2005, any occurrence of TB in the free-living animals inhabiting this area might pose a real threat to local livestock and result in the loss of disease-free status. The aim of the study was to describe the occurrence of tuberculosis in the wildlife of the Bieszczady Mountains and determine the microbiological and molecular characteristics of any cultured strains. Lymph node samples were collected for analysis from 274 free-living animals, including European bison, red foxes, badgers, red deer, wild boar and roe deer between 2011 and 2017. Löwenstein–Jensen and Stonebrink media were used for culture. Molecular identification of strains was performed based on hsp65 sequence analysis, the GenoType®MTBC (Hain Lifescience, Germany) test, spoligotyping and MIRU-VNTR analysis. Results Mycobacterium caprae was isolated from the lymph nodes of 21 out of 55 wild boar (38.2%; CI 95%: 26.5%, 51.4%) and one roe deer. Since 2014, no new TB cases have been reported in the Bieszczady European bison population. Conclusions The identification of TB in wild boar in the Bieszczady is an alarming phenomenon, which requires further investigation. The Bieszczady mountains are a precious, unique area, home to many protected species. However, it is also the only area in Poland where TB cases have been reported in free-living animals. The occurrence of TB in wild boar inhabiting this area might pose a real threat to local livestock and many of the protected species (for example European bison that can share feeding places with wild boar). Given this situation, ongoing monitoring of the prevalence of TB should be conducted, and protective measures should be considered.

Introduction Bacillus Calmette-Guerin (BCG) is the only tuberculosis vaccine available and although it has been routinely used for more than 80 years, its protective effect varies depending on the age and the form of tuberculosis. Due to the close analogy between the vaccine strain and other species of the Mycobacterium tuberculosis complex, molecular methods are recommended for differential diagnosis of post-BCG complications. The aim of the study was to assess usefulness of molecular methods in diagnosis of post-BCG vaccine adverse events (VAEs). Material and methods M. tuberculosis complex strains obtained in 2011-2017 from 68 ill children were subjected to molecular analysis. Results Molecular analysis of 68 strains showed 100% agreement between the results in the GenoType MTBC method and the multiplex PCR method. For the strains isolated from 45 patients with suspected VAE, M. bovis BCG was obtained, whereas the strains isolated from the remaining 23 children were identified as M. tuberculosis. The analysis confirmed the close relationship between the result of identification and the type of material as well as the patient's age. Conclusions The use of genetic methods enables quick and detailed diagnostics of infections caused by M. bovis BCG, which allows for the confirmation or exclusion of VAE.

Interferon gamma releasing assays (IGRAs) are extensively used in the diagnosis of latent tuberculosis infections. Comparing to tuberculin skin test (TST) they lack false positive results in the populations vaccinated with BCG, and in most non-tuberculous mycobacteria (NTM) infections. Nevertheless, Mycobacterium kansasii, Mycobacterium marinum, and Mycobacterium szulgai may induce positive IGRAs due to RD1 homology with Mycobacterium tuberculosis. The aim of the study was to investigate the possible influence of NTM respiratory isolates on the results of IGRAs. 39 patients (23 females and 16 males) of median age 61 years, with negative medical history concerning tuberculosis, entered the study. Identification of NTM was performed using the niacin test and molecular method GenoType CM test (Hain Lifescience). QFT-Plus was performed in 17 patients, T-SPOT-Tb - in 23 patients. Chest X-rays and a high-resolution computed tomography of the chest have been reviewed by the experienced radiologist blinded to the results of IGRAs, in search of past tuberculosis signs. Positive IGRAs results were obtained in three out of 39 patients (8%): 22% of patients with M. kansasii isolates and 18% of patients with radiological signs on HRCT that might be suggestive of past tuberculosis. Positive IGRAs correlated with radiological signs suggestive of past tuberculosis (r = 0.32, p = 0.04), and on the borderline with isolation of M. kansasii (r = 0.29, p = 0.06). These findings may suggest that a positive IGRAs result, in our material, could depend mostly on asymptomatic past Tb infection. The cross-reactivity of M. kansasii isolates with IGRAs was less probable; nevertheless, it requires further investigations. Interferon gamma releasing assays (IGRAs) are extensively used in the diagnosis of latent tuberculosis infections. Comparing to tuberculin skin test (TST) they lack false positive results in the populations vaccinated with BCG, and in most non-tuberculous mycobacteria (NTM) infections. Nevertheless, Mycobacterium kansasii, Mycobacterium marinum, and Mycobacterium szulgai may induce positive IGRAs due to RD1 homology with Mycobacterium tuberculosis. The aim of the study was to investigate the possible influence of NTM respiratory isolates on the results of IGRAs. 39 patients (23 females and 16 males) of median age 61 years, with negative medical history concerning tuberculosis, entered the study. Identification of NTM was performed using the niacin test and molecular method GenoType CM test (Hain Lifescience). QFT-Plus was performed in 17 patients, T-SPOT-Tb – in 23 patients. Chest X-rays and a high-resolution computed tomography of the chest have been reviewed by the experienced radiologist blinded to the results of IGRAs, in search of past tuberculosis signs. Positive IGRAs results were obtained in three out of 39 patients (8%): 22% of patients with M. kansasii isolates and 18% of patients with radiological signs on HRCT that might be suggestive of past tuberculosis. Positive IGRAs correlated with radiological signs suggestive of past tuberculosis (r = 0.32, p = 0.04), and on the borderline with isolation of M. kansasii (r = 0.29, p = 0.06). These findings may suggest that a positive IGRAs result, in our material, could depend mostly on asymptomatic past Tb infection. The cross-reactivity of M. kansasii isolates with IGRAs was less probable; nevertheless, it requires further investigations.

Introduction: An increasing incidence rate of respiratory isolates of non-tuberculous mycobacteria (NTM) has been noted recently in most European countries as well as in the US. Despite many publications, there is no consensus concerning the importance of different factors in promoting NTM lung disease (NTMLD). The aim of the present retrospective study was to analyse patients with positive NTM respiratory isolates in search of factors predisposing to NTMLD. Material and methods: 73 patients, 23 males, 50 females, median age 62.2 years, in whom NTM have been cultured from respiratory specimen (sputum and/or bronchial washings), in the period 2010–2015, entered the study. Results: NTMLD (according to ATS/IDSA) has been recognised in 36 patients, airways colonisation by NTM—in 37 patients. NTMLD was diagnosed more often in the patients infected with M. kansasii, M. abscessus and M. avium/M. intracellulare comparing to those infected with M. xenopi, M. gordonae and M. fortuitum (p < 0.0001). The proportion of females to males was significantly higher in the NTMLD group comparing to the colonisation group (p < 0.007). Previous tuberculosis or mycobacteriosis were noted significantly more frequently in the group of patients with NTMLD comparing to the colonisation group (28% vs. 8%, p = 0.038). Univariate regression analysis revealed M. kansasii, female gender, and previous tuberculosis or mycobacteriosis as significant predictors of NTMLD. Conclusions: The risk factors of NTMLD recognition in the presented group of patients were the following: female gender, M. kansasii isolation, as well as past tuberculosis or mycobacteriosis.

Introduction: During the last decades the prevalence of NTM infections has increased, especially in developed countries. The aim of the study was to provide an overview on all NTM isolated from clinical samples in Poland between 2013 and 2017. Material and methods: The study comprised 2799 clinical specimens, mostly respiratory accessed in the reference laboratory of National Tuberculosis and Lung Diseases Research Institute in Warsaw and in the Wielkopolska Center of Pulmonology and Thoracic Surgery, Poland, 2013–2017. Results: During the study period 35 species of NTM were isolated . The number of isolates increased almost 1.6-fold: from 420 in 2013 to 674 in 2017. M. kansasii , M. avium , M. xenopi , M. gordonae and M. intracellulare were the most common species. This NTM pattern was rather stable over the time. If the aggregated amount of all MAC species was taken into account they dominated over M. kansasii from 2015. M. avium and M. intracellulare were more often isolated from women, while M. kansasii , M. gordonae and M. xenopi predominated in men. Men and women were infected almost with the same frequency. In older patients 65+ women were in majority, quite opposite to those aged 25 to 64 years. Conclusion: In Poland, like in other countries increased the frequency of isolated NTM. M. kansasii and M. avium were the most frequently identified species from clinical samples. Men and women were infected with NTM with the same frequency.

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.

Non-tuberculous mycobacteria (NTM) are increasingly a cause of human respiratory tract colonization and mycobacterial lung disease (NTM-LD), especially in patients with chronic lung diseases. The aim of the present study was to find the factors predictive of NTM-LD in patients with obstructive lung diseases and NTM respiratory isolates. A total of 839 isolates of NTM, obtained from 161 patients between 2010 and 2020 in a single pulmonary unit, have been retrospectively reviewed. Of these isolates, 73 concerned 36 patients with obstructive lung diseases (COPD-26, asthma-3, COPD/asthma overlap syndrome-7). NTM-LD was recognized according to the American Thoracic Society (ATS) and the Infectious Diseases Society of America (IDSA) criteria in 17 patients, colonization in 19. Lower BMI, elevated body temperature on admission, infiltrative/cavitary lesions on chest CT, and NTM species other than Mycobacterium gordonae were the significant predictors of NTM-LD recognition. Based on the above-mentioned predictive factors, an original scoring system was implemented. The diagnostic utility of the scoring system was higher than that of single parameters. We conclude that NTM-LD prediction in patients with obstructive lung diseases and positive respiratory isolates is difficult. A scoring system based on clinical, radiological and microbiological characteristics was capable of facilitating the differential diagnosis, but it needs further validation in a larger study group.

Summary Introduction. The main causes of drug-resistant TB is the use of improper treatment regimens, and incorrect diagnosis of patients. The rapid identification of drug resistance strain is an important challenge to ensure a rapid and adequate therapy of tuberculo- sis and to limit the dissemination of multidrug resistant strains. Isoniazid (INH) and ri- fampicin (RMP) are the main antituberculosis drugs, used in the treatment regimen rec- ommended by the WHO. The occurrence of resistance to these two basic medicines are considered to be one of several causes of treatment failure in patients with tuberculosis. Aim. The aim of this study was to compare the results of resistance to isoniazid and rifampicin obtained with the molecular and the phenotypic method. Material and methods. The MTBDR plus assay was performed on 60 strains Mycobac- terium tuberculosis complex, and the results were compared with the results of conven- tional drug susceptibility testing (Bactec MGIT 960). Results. In the analyzed group of 60 strains of mycobacterium tuberculosis, mutations in the rpoB gene were detected in 40 of 42 strains of M. tuberculosis phenotypically resist- ant to RMP (92.9%). In the group of 51 M. tuberculosis strains phenotypically resistant to INH, mutations in the katG gene and inhA gene were found in 46 (90.2%). The sensitivity, specificity and positive and negative predictive values of the MTBDR assay were respec- tively 82.9, 94.7, 97.1 and 72% for multidrug-resistant TB (MDR-TB). Conclusions. The GenoType MTBDR plus assay is a molecular test which detecting the most common mutations in strains resistant to RMP and INH. However the test does not detect all mutations associated with resistance to isoniazid and rifampicin, therefore, the results of molecular must be confirmed by phenotype.

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.

It is estimated that one third of the world’s population have latent tuberculosis infection and that this is a significant reservoir for future tuberculosis cases. Most cases occur within two years following initial infection. The identification of individuals with latent tuberculosis infection is difficult due to the lack of an ideal diagnostic assay and incomplete understanding of latent infection. Currently, there are three tests: the oldest tuberculin skin test, T-SPOT.TB and the latest QuantiFERON-Plus for the detection of Mycobacterium tuberculosis infection. The interpretation of the test results must be used in the conjunction with a patient’s epidemiological history, risk assessment, current clinical status, radiography and microbiological methods to ensure accurate diagnosis.

The aim of this study was to assess whether animal tuberculosis (TB) is transmitted between free-living European bison (Bison bonasus caucasicus), wild boars (Sus scrofa), and protected carnivores such as grey wolves (Canis lupus), brown bears (Ursus arctos), and Eurasian lynx (Lynx lynx) in the Bieszczady Mountains in Southern Poland. Results of animal studies suggest that TB transmission from bison or wild boars to grey wolves is possible. These are the first described cases where Mycobacterium caprae was detected in samples collected from grey wolves.

Mikobakteriozy to choroby wywoływane przez prątki niegruźlicze (NTM), zwane również prątkami atypowymi. Mikobakterie są dość powszechne w naszym środowisku, głównie w glebie i wodzie. Mogą one kolonizować drogi oddechowe, przewód pokarmowy i układ moczowo-płciowy człowieka. U osób z przewlekłymi chorobami płuc oraz z obniżoną odpornością mogą być przyczyną pogorszenia przebiegu choroby podstawowej. Obecność NTM w plwocinie pacjentów z mukowiscydozą (CF) stanowi poważny dylemat diagnostyczny, ponieważ może wskazywać na przejściowe zanieczyszczenie, kolonizację lub zakażenie. Wyniki badań epidemiologicznych wskazują, że większość zakażeń prątkami niegruźliczymi u chorych na mukowiscydozę jest wywołana przez Mycobacterium abscessus i Mycobacterium avium. Diagnostyka i leczenie mikobakteriozy płuc u chorych na CF wymagają również indywidualnego podejścia do każdego chorego. W niniejszej pracy opisano 3 przypadki mikobakteriozy płuc o różnym przebiegu klinicznym u chorych na mukowiscydozę oraz trudności w ich diagnostyce i leczeniu.

Wstęp: W ostatnich latach w większości krajów Europy i w Stanach Zjednoczonych jest notowany wzrost częstości zakażeń układu oddechowego prątkami niegruźliczymi. Nie ma jednak zgodności opinii na temat wpływu różnych czynników na rozwój mykobakteriozy płuc w tej grupie chorych. Celem niniejszej pracy retrospektywnej było poszukiwanie czynników ryzyka mykobakteriozy płuc, w grupie chorych, u których wyhodowano prątki niegruźlicze z dróg oddechowych. Materiał i metody: Badaną grupę stanowiło 73 chorych, 23 mężczyzn i 50 kobiet, mediana wieku wyniosła 62.2 roku, z dodatnimi wynikami posiewów plwociny/wydzieliny oskrzelowej w kierunku prątków niegruźliczych, uzyskanymi w latach 2010–2015. Wyniki: Mykobakteriozę płuc (według American Thoracic Society/Infectious Diseases Society of America) rozpoznano u 36 chorych, kolonizację dróg oddechowych—w 37 przypadkach. Mykobakteriozę rozpoznawano istotnie częściej u chorych zakażonych M. kansasii, M. abscessus i M. avium/M. intracellulare w porównaniu z chorymi zakażonymi M. xenopi, M. gordonae i M. fortuitum (p < 0.0001). Wśród chorych na mykobakteriozę, w porównaniu z grupą z kolonizacją prątkami niegruźliczymi, było istotnie więcej kobiet niż mężczyzn (p < 0.007) oraz więcej chorych z wywiadem przebytej gruźlicy płuc lub mykobakteriozy (odpowiednio 28% i 8%, p = 0.038). Wykazano, że zakażenie M. kansasii, płeć żeńska oraz przebyta gruźlica lub mykobakterioza były istotnymi czynnikami predykcyjnymi aktualnego rozpoznania mykobakteriozy płuc. Wniosek: W badanej grupie chorych ryzyko zachorowania na mykobakteriozę było istotnie większe u kobiet, osób zakażonych M. kansasii oraz z przebytą gruźlicą lub mykobakteriozą płuc w przeszłości.

Wstęp: W kilku ostatnich dekadach obserwuje się wzrost izolacji prątków niegruźliczych (NTM, nontuberculous mycobacteria), zwłaszcza w krajach rozwiniętych. Celem pracy była analiza wszystkich gatunków prątków niegruźliczych izolowanych z materiałów biologicznych od pacjentów w Polsce, w latach 2013–2017. Materiał i metody: Badaniem objęto 2799 izolatów, pochodzących głównie z układu oddechowego, które oceniano w referencyjnym laboratorium Instytutu Gruźlicy i Chorób Płuc w Warszawie oraz w Wielopolskim Centrum Pulmonologii i Chirurgii Klatki Piersiowej. Wyniki: W badanym okresie izolowano 35 różnych gatunków NTM. Liczba izolatów wzrosła prawie 1,6-krotnie: od 420 w 2013 roku do 674 w 2017 roku. Najczęściej występowały: M. kansasii, M. avium, M. xenopi, M. gordonae i M. intracellulare. Dominacja tych gatunków NTM w prawie niezmienionej kolejności utrzymywała się przez cały okres. Natomiast przy rozpatrywaniu MAC (M. avium complex) jako całości—od 2015 roku kompleks ten dominował nad M. kansasii. M. avium i M. intracellulare częściej izolowano od kobiet, podczas gdy M. gordonae i M. xenopi od mężczyzn. Nie stwierdzono różnic w częstości występowania NTM u mężczyzn i kobiet. Wśród pacjentów po 65. roku życia dominowały kobiety, zaś u tych pomiędzy 25. a 64. rokiem życia—mężczyźni. Wnioski: W Polsce, podobnie jak w innych krajach, zwiększa się częstość izolacji NTM. Najczęściej izolowanymi gatunkami były: M. kansasii i M. avium. Prątki niegruźlicze występowały u kobiet i mężczyzn z podobną częstością.

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.

N-acetylotransferazy (NAT; EC2.3.1.5) biorą czynny udział w metabolizowaniu leków i toksyn środowiskowych. Katalizują przeniesienie grupy acetylowej z acetylokoenzymu A do terminalnej grupy aminowej aryloamin, arylohydrazyn i niektórych amin heterocyklicznych. Enzymy arylamino N-acetylotransferazy zostały zidentyfikowane u wielu organizmów eukariotycznych i prokariotycznych. Polimorfizm genetyczny N-acetylotransferaz powoduje powstanie enzymów o zmienionej sekwencji aminokwasowej, która przyczynia się do obniżenia ich aktywności i stabilności. Występowanie polimorfizmu w aktywności enzymu N-acetylotransferazy 2 jest przyczyną występowania dwóch odmiennych fenotypowo grup: wolnych i szybkich acetylatorów. Szybkość metabolizowania leków oraz związków kancerogennych zależnych od N-acetylotransferaz wpływa na skuteczność i efekt toksyczny tych leków oraz może mieć związek ze zwiększonym ryzykiem wystąpienia niektórych jednostek chorobowych. Badania epidemiologiczne sugerują, że polimorfizm N-acetylotransferaz może mieć wpływ na ryzyko zachorowania na raka pęcherza moczowego, jelita grubego, piersi, głowy i szyi oraz płuc.

Isoniazid (INH), a key agent in the treatment of tuberculosis (TB), is metabolized primarily by the genetically polymorphic N-acetyltransferase 2 (NAT2) enzyme. Patients treated with INH can be classified as fast, intermediate, and slow acetylators. The objective of this study was to explore the relationship between NAT2 genotypes and the serum concentrations of INH. Blood samples from 130 patients were taken for the analysis, and plasma INH concentrations were determined by using the high-performance liquid chromatography (HPLC) technology. Acetylation genotype was determined on genomic DNA by using an allele-specific polymerase chain reaction-restriction fragment length polymorphism (PCR–RFLP) assay. Once the NAT2 genotypes were established, patients were classified into three categories: fast, intermediate, and slow acetylators. Of the 130 patients studied, 84 (64.6%) were slow, 39 (30%) were intermediate, and 7 (5.4%) were fast acetylators. Analysis of INH concentrations in the blood of patients receiving the approximate doses of the drug revealed that, at the time intervals examined, the average concentration of INH was 2- to 7-fold higher among slow acetylators compared to fast and intermediate acetylators.Conclusion. Determining mutations in the NAT2 gene enabled the identification of the INH acetylation type in patients and the genotyping results were consistent with the phenotype determined by methods of measurement of drug bioavailability.

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.

Wstep: Mikobakteriozy są chorobami rzadkimi, rozpoznawanymi glownie u osob z grup ryzyka. Wśrod plucnych czynnikow ryzyka nie wymieniono dotychczas nadciśnienia plucnego. Celem pracy byla analiza obrazu klinicznego i przebiegu mikobakteriozy pluc, ktorą rozpoznano w ośrodku autorow pracy w latach 2002–2012 u chorych na nadciśnienie plucne zakrzepowo-zatorowe (CTEPH) i idiopatyczne (IPAH), oraz proba określenia czynnikow sprzyjających zachorowaniu. Material i metody: Badaną grupe stanowilo 13 chorych — 10 z CTEPH i 3 z IPAH. Nadciśnienie plucne potwierdzono metodą inwazyjną. Mediana średniego ciśnienia w tetnicy plucnej w okresie rozpoznania mikobakteriozy wynosila 49 mm Hg (39–65 mm Hg). Mikobakterioze pluc rozpoznano zgodnie z kryteriami Amerykanskiego Towarzystwa Chorob Pluc z 2007 roku. Wyniki: Czynnikiem etiologicznym byl u wiekszości chorych gatunek M. kansasii. Najczestszymi objawami mikobakteriozy pluc byly nasilenie duszności i produktywny kaszel. W tomografii komputerowej klatki piersiowej z opcją naczyniową w 7 przypadkach stwierdzono zageszczenia miązszowe z rozpadem, w 6 — jamy otoczone drobnymi guzkami. U wszystkich chorych na CTEPH zmiany związane z mikobakteriozą pojawily sie na obszarach o upośledzonej perfuzji, ale bez widocznych zmian pozawalowych. Wszyscy chorzy otrzymali leczenie przeciwprątkowe, uzyskano poprawe w 12/13 przypadkow. Do kwietnia 2014 roku 7 chorych zmarlo z powodu ciezkiej prawokomorowej niewydolności serca, nie obserwowano zgonow w przebiegu mikobakteriozy. Wnioski: Nowe plucne zmiany miązszowe z rozpadem u chorych na CTEPH i na IPAH z towarzyszącym produktywnym kaszlem powinny sklonic do diagnostyki w kierunku mikobakteriozy pluc. W CTEPH niepokoj budzą szczegolnie ogniska rozpadu, ktorym nie towarzyszą nowe skrzepliny w naczyniach doprowadzających. Czynnikami sprzyjającymi rozwojowi mikobakteriozy pluc są: wysokie nadciśnienie plucne (CTEPH i IPAH) oraz obnizona perfuzja (CTEPH).

Mycobacterial lung disease is caused by nontuberculous mycobacteria (NTM), also known as atypical mycobacteria. NTM are widely distributed in the environment, particularly in soil and water. Although generally of low pathogenicity to humans, NTM can affect patients with underlying chronic lung diseases, such as cystic fibrosis, bronchiectasis, pneumoconiosis, or healed tuberculosis. Some patients with cystic fibrosis (CF) have disease progression due to NTM, others can have NTM cultured intermittently from respiratory specimens without a significant decline in lung function. Identifying which patients will worsen from NTM and therefore need treatment remains difficult because of the similarity of symptoms in CF and NTM lung disease. The most common species of NTM isolated in CF patients are Mycobacterium avium complex (MAC) and Mycobacterium abscessus. In this paper, we present three different cases of mycobacterial lung disease in patients with cystic fibrosis.

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.

The N-acetyltransferases (NAT; E.C.2.3.1.5) are involved in the metabolism of drugs and environmental toxins. They catalyse the acetyl transfer from acetyl coenzyme A to an aromatic amine, heterocyclic amine, or hydrazine compound. NAT homologues are present in numerous species from bacteria to human. Sequence variations in the human NAT1 and NAT2 result in the production of NAT proteins with variable enzyme activity or stability, leading to slow or rapid acetylation. Therefore, genetic polymorphisms in NAT1 and NAT2 influence drug metabolism and drug-related toxicity. Epidemiological studies suggest that the NAT1 and NAT2 acetylation polymorphisms modify the risk of developing cancers of the urinary bladder, colorectal, breast, head and neck, and lung.

Individual's risk of developing lung cancer depends not only on exposure to tobacco smoke, but also on the activity of enzymes involved in the activation or deactivation of carcinogens. Arylamine N-acetyltransferase (EC 2.3.1.5) is an enzyme involved in biotransformation of xenobiotics, mainly aromatic and heterocyclic amines and hydrazines. The different acetylation phenotypes within a population are derived from mutations in the NAT 2 gene. These mutations influence the activity (specifically resulting in high or low activity) of the NAT enzyme. Some authors have demonstrated lung cancer predisposing role of slow acetylator phenotype, whereas other reported increased lung cancer risk for fast acetylators or neutral effect of the NAT2 polymorphism. The aim of this preliminary report was to determine the NAT2 gene polymorphism in patients with lung cancer.39 patients with inoperable lung cancer (29 - NSCLC and 10 - SCLC), median age 59 years (42- -72) entered the study. Acetylation genotype was determined in the genomic DNA using an allele-specific polymerase chain reaction. We investigated four genetic mutations, C481T, G590A, A803G i G857A, of the gene NAT2.There were 10 different NAT2 genotypes among the 39 patients. Fourteen patients with a NAT2*2 4/4, *4/5, *4/6 and *4/7 were classified as fast acetylators; and 25 patients with a NAT2*5/5, *5/6, *5/7, *6/6, *6/7 or *7/7 genotype were classified as slow acetylators. Among the 10 patients with SCLC - 4 were fast acetylators, and among 29 patients with NSCLC dominated slow acetylation type found in 19 patients (genotypes NAT2 *5/5 and NAT2 *5/6).Among patients with small cell lung cancer, there was no predominance of genotype of acetylation, whereas among patients with non-small cell lung cancer predominated NAT2*5/5 and NAT2*5/6 genotypes (slow acetylators).

Wstęp: Indywidualne ryzyko zachorowania na raka płuca zależy nie tylko od ekspozycji na dym tytoniowy, ale również od aktywności enzymów biorących udział w aktywacji lub deaktywacji substancji rakotwórczych. Arylamino N-acetylotransferazy (EC 2.3.1.5) są enzymami biorącymi udział w biotransformacji ksenobiotyków, amin aromatycznych i heterocyklicznych oraz hydrazyn. Zaobserwowane różnice w aktywności enzymu i szybkości metabolizowania substancji zależnych od N-acetylotransferazy 2 (NAT2) powiązano z polimorfizmem genu kodującego ten enzym. Niektórzy autorzy wskazują na wolny typ acetylacji, jako predysponujący do wystąpienia raka płuca, podczas gdy inni wykazują brak wpływu polimorfizmu NAT2 lub większe ryzyko raka płuca wśród szybkich acetylatorów. Celem pilotażowego badania była ocena polimorfizmu genu NAT2 umożliwiającego określenie typu acetylacji u chorych na raka płuca. Materiały i metody: Badaną grupę stanowiło 39 chorych na nieoperacyjnego raka płuca (29—rak niedrobnokomórkowy, 10—rak drobnokomórkowy), mediana wieku wynosiła 59 lat (42–72 lata). Do badania pobierano 5 ml krwi. Genotyp NAT2 został określony na podstawie identyfikacji czterech mutacji, C481T, G590A, A803G i G857A. Wyniki: W przebadanej grupie 39 chorych zidentyfikowano występowanie 10 różnych genotypów NAT2. Czternastu chorych z genotypami NAT2 *4/4, *4/5, *4/6 i *4/7 zostało sklasyfikowanych jako szybcy acetylatorzy a 25 z genotypami NAT2 *5/ 5, *5/6, *5/7, *6/6, *6/7 lub *7/7 jako wolni acetylatorzy. Wśród 10 chorych na DRP — 4 chorych to szybcy acetylatorzy, zaś wśród 29 chorych na NDRP dominował wolny typ acetylacji stwierdzony u 19 chorych (genotypy NAT2*5/5 i NAT2*5/6). Wnioski: Wśród chorych na drobnokomórkowego raka płuca nie stwierdzono dominacji określonego genotypu acetylacji, natomiast wśród chorych na niedrobnokomórkowego raka płuca przeważali pacjenci z genotypami NAT2*5/5 i NAT2*5/6 (wolni acetylatorzy).

Introduction: Correctional facilities are recognised “breeding ground” for infectious diseases. As The World Health Organization reported, the incidence of infectious diseases in prison’s population is 10–100 times higher than in general population. The incidence of tuberculosis among correctional inmates in Poland in 2008 was 270/100,000, that is around 10 times higher than among non-prisoners. Materials and methods: The study included 57 M. tuberculosis isolates from patients in Polish prisons in 2004–2008 (5% of all diagnosed TB patient in Polish prisons 2004–2008). Primary isolation was performed with Löwenstein-Jensen (L-J) medium, species identification was done with the niacin test and gene probes test. Bacterial DNA was extracted from the L-J medium slants with the cetyltrimethylammonium bromide (CTAB) method. Mycobacterium tuberculosis strains were analyzed with two methods: screening for epidemiological discrimination of M. tuberculosis—spoligotyping and highthroughput—MIRU/VNTR. Results: Isolates that are grouped in clusters (33 isolates) were analyzed by means of MIRU/VNTRs. In MIRU/VNTRs all strains showed different genetic patterns. Most isolates of the prisoners were grouped into two clusters: T1 53 and H3 50. Conclusions: 1. MIRU/VNTR is a high-throughput method. 2. MIRU/VNTR is a promising method to diagnose TB transmission in Polish jails. 3. To identify the probable source of transmission, molecular analysis of strains from patients of the general population is needed.

The diagnosis of latent tuberculosis infection (LTBI) is currently based on the century-old tuberculin skin test (TST). However a positive reaction can result from infection by Mycobacterium tuberculosis, BCG vaccination or cross-reaction with nontuberculous mycobacteria. T-SPOT.TB assay is a new test to diagnose tuberculosis infection by measuring in vitro T-cell interferon gamma release in response to two Mycobacterium tuberculosis-specific antigens: ESAT-6 and CFP 10.T-SPOT.TB assay has been performed on whole blood samples (n = 137) from March to September 2010. A tuberculin skin test result was available for 96 of participants. A positive TST result was considered if the induration was 10 mm or more.Of the 137 patients tested, T-SPOT.TB assay results were positive in 37 (27%), negative in 98 (71.5%) and indeterminate in only 2 (1.5%) persons. We analyzed T-SPOT.TB and TST results in the 96 patients for whom both test were available. Concordance between T-SPOT.TB and TST results (10 mm skin reaction interpreted as positive) was 79%. Fifteen (15.6%) patients had a positive TST result and a negative T-SPOT.TB and 5 (5.2%) patients had a negative TST result and a positive T-SPOT.TB. We observed good correlation between positive T-SPOT.TB results and the size of induration ≥ 15 mm in TST results.T-SPOT.TB offers a more accurate approach than TST for identification tuberculosis infection. The study shows that the test T-SPOT.TB is a good diagnostic tool in identifying persons with tuberculosis infection. For full confirmation of this assessment, it is necessary to examine more cases.

Introduction: Diagnostics of latent tuberculosis infection (LTBI) has been based on a century-old tuberculin skin test (TST). However, a positive reaction can result not only from infection with Mycobacterium tuberculosis, but also from BCG vaccination or cross-reaction with nontuberculous mycobacteria. T-SPOT.TB assay is a new test to diagnose tuberculosis infection by measuring in vitro T-cell interferon-gamma release in response to two Mycobacterium tuberculosis-specific antigens: ESAT-6 and CFP-10. Material and methods: T-SPOT.TB assay was performed on samples of whole blood (n = 137) from March to September 2010. Tuberculin skin test was carried out in 96 participants. A positive TST result was considered to be an induration of 10 mm or more. Results: Of the 137 patients tested, T-SPOT.TB assay results were positive in 37 (27%), negative in 98 (71.5%) and indeterminate in only 2 (1.5%) persons. We analyzed T-SPOT.TB and TST results in 96 patients who were subjected to both tests. Concordance between T-SPOT.TB and TST results (a 10-mm skin reaction interpreted as positive) was 79%. Fifteen (15.6%) patients had a positive TST result and a negative T-SPOT.TB, and 5 (5.2%) patients had a negative TST result and a positive T-SPOT.TB. We observed a good correlation between positive T-SPOT.TB results and the diameter of induration of ≥15 mm in TST results. Conclusions: T-SPOT.TB offers a more accurate approach than TST in the identification of tuberculosis infection. The study showed that the test T-SPOT.TB is a good diagnostic tool in identifying persons with tuberculosis infection. For a full confirmation of this assessment, it is necessary to examine more cases.

The aim of this study was estimation of usefulness of two molecular methods for Mycobacterium tuberculosis typing in the epidemiological research of tuberculosis (TB). We determined molecular patterns of M. tuberculosis strains isolated from 66 patients, members of 29 families living in 9 voivodeships of Poland. We also analysed drug susceptibility of the strains as well as some demographic characteristics of the patients.The genotype analysis of the 66 clinical isolates was performed by using spoligotyping and IS6110-Mtb1/Mtb2 PCR.Of the 29 families examined in this study, in 23 each family member was infected with the same M. tuberculosis strain. Three drug-resistant strains and two members of the Beijing family were identified.We found that strains within each of the 23 families had the same genetic patterns, whereas those of the strains identified in the rest 6 families were different. Among those 6 families, in 3 differentiation of the strains was obtained with both spoligotyping and IS6110-Mtb1/Mtb2 PCR analysis, while in another 3 only with spoligotyping method.Based on the results from this study, the two genotyping methods used were demonstrated as an efficient approach for investigating the epidemiological relatedness of TB cases.