Nasal carriage of various staphylococcal species in small ruminant lentivirus-infected asymptomatic goats.

The study was carried out in Polish goat population to estimate the prevalence of the nasal cavity infection with various staphylococcal species including methicillin-resistant Staphylococcus aureus(MRSA), investigate the potential permissive role of small ruminant lentivirus (SRLV) infection and determine the level of clonality of S. aureus nasal isolates. Nasal swabs and blood samples were collec-ted from 1300 clinically healthy adult goats from 21 Polish goat herds. Blood samples were serological-ly screened for SRLV. Staphylococci were isolated from nasal swabs and identified using classical microbiological methods, MALDI-TOF, multiplex-PCR, and their clonality was assessed using PFGE. Antimicrobial resistance was determined on the basis of minimum inhibitory concentration and by demonstration of the presence of the mecA gene encoding the multiplex-PCR PBP2a protein and of the five main types of staphylococcal cassette chromosome mec. The apparent prevalence of staphylococ-cal and S. aureus infection of the nasal cavity was 29.1% (CI 95%: 26.9%, 31.5%) and 7.3% (CI 95%: 6.1%, 8.8%), respectively. No relationship was found between the SRLV-infection and the presence of any staphylococcal species including S. aureus (p=0.143). Only 9.8% of S. aureus isolates were resistant to amoxicillin/clavulanic acid and 5.9% to chloramphenicol and ciprofloxacin. All tested isolates proved to be phenotypically and genotypically sensitive to methicillin, which yielded the appar-ent prevalence of MRSA of 0% (CI 95%: 0%, 7.0%). S. aureus isolates show high genetic similarity within goat herds, however vary considerably between herds. Goats do not appear to be an important source of S. aureus for humans in Poland.

Antineutrophil cytoplasmic antibodies and their relationship with disease activity and presence of staphylococcal superantigens in nasal swabs in patients having granulomatosis with polyangiitis: results of a study involving 115 patients from a single center.

OBJECTIVE: Antineutrophil cytoplasmic antibodies (ANCAs) are considered a risk factor for granulomatosis with polyangiitis (GPA) exacerbation, especially when staphylococcal superantigens (SAgs) are present in nasal swabs. Their role in monitoring disease activity remains controversial. This study determined the relationship of ANCAs with disease activity and presence of SAgs in GPA patients. METHODS: Among a total of 115 GPA patients hospitalized in the period 2009-2016, we investigated the presence of SAgs and ANCA concentration. Blood samples and nasal swabs were taken at each visit (referred further to as episodes). Disease activity was assessed using the Birmingham Vasculitis Activity Score (BVAS). RESULTS: We analyzed 362 episodes. ANCAs were detected in 215 (59.4%), while SAgs were detected in 126 (34.8%) episodes. We found a significant correlation between the presence of ANCAs and disease activity (p = 0.0032), as well as between their level and GPA severity (r = 0.25363, p = 0.000001). We also determined that an ANCA values ≥ 138 Ru/ml were an indicator of active disease with high specificity and low sensitivity (84.4% and 37.3%, respectively). The relationship between ANCA presence and the presence of SAgs was not confirmed; however, when SAgs were analyzed based on the different types, ANCA levels were found to be significantly higher in the group with SAg type B (p = 0.031). CONCLUSIONS: There was no detectable evidence for the association between ANCA level and the presence of SAgs. Although monitoring ANCA levels as a marker of disease activity may be clinically relevant, GPA management cannot proceed on the basis of ANCA levels alone. Key Points • ANCA concentration usually correlates with GPA activity, although in half of patients, ANCAs persist despite effective treatment and clinical remission. • ANCA values of 138 Ru/ml seem to be an indicator of active disease with high specificity, but low sensitivity. • Although there is a relevance for ANCA monitoring as a marker of disease activity, GPA management cannot be based on ANCA levels alone. • The suspected clinical correlation between ANCA formation and SAg presence in nasal swabs is not obvious and requires further investigations.

Bovine tuberculosis outbreak in farmed American bison (Bison bison) in Poland.

Poland has been an officially bovine tuberculosis (bTB) free country for the last seven years. The problem currently observed is the increasing number of new cases of bTB in wild species, kept in a farmed herd and free-living herd: European bison (Bison bonasus), wild boar (Sus scrofa), wolves (Canis lupus) and red deer (Cervus elaphus). This article presents the case of Mycobacterium caprae transmission to an American bison (Bison bison) herd kept on a private farm in Eastern Poland.

A close-up on the epidemiology and transmission of multidrug-resistant tuberculosis in Poland.

Multidrug-resistant tuberculosis (MDR-TB) poses a serious challenge to the global control of the disease. The purpose of this study was to characterize MDR-TB patients from Poland and to determine the extent of MDR-TB disease attributable to recent transmission. The study included all 46 patients diagnosed with MDR-TB in Poland in 2004 and followed up for 6 years (until 2011). For each patient, sociodemographic and clinical characteristics, treatment outcomes, and bacteriological data were collected by the review of medical and laboratory records. Mycobacterium tuberculosis isolates from all patients were characterized using spoligotyping, mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing, IS6110 restriction fragment length polymorphism (RFLP) analysis, and sequencing analysis of drug resistance-associated loci (katG, mabA-inhA, rpoß, rpsL, and embB). The majority of patients were male (86.9%), 40-64 years of age (60.8%), with a history of TB treatment (84.8%), and producing smear-positive sputa (86.9%). Twenty-two (47.8%) patients suffered from concomitant diseases and 28 (60.8%) were alcohol abusers. Treatment outcome assessment revealed that 8 (17.4%) patients were cured or completed therapy, while 15 (32.6%) died of TB, 11 (23.9%) defaulted, 8 (17.4%) failed, and 1 (2.2%) was transferred and lost to follow-up. Upon genotyping, 10 (21.7%) isolates were allocated in four clusters. These were further subdivided by mutational profiling. Overall, in 6 (13%) patients, MDR-TB was a result of recent transmission. For 4 (8.7%) of these patients, a direct epidemiological link was established. The study shows that the transmission of MDR-TB occurs at a low rate in Poland. Of urgent need is the implementation of a policy of enforced treatment of MDR-TB patients in Poland.

Characterisation of pyrazinamide-resistant Mycobacterium tuberculosis strains isolated in Poland and Germany.

BACKGROUND: Pyrazinamide (PZA) is an important first-line anti-tuberculosis drug that is generally administered with isoniazid, rifampicin, ethambutol and streptomycin. OBJECTIVE: To analyse the correlation between phenotypic resistance to PZA and genotype to find out whether the great diversity in pncA mutations is epidemiologically useful in tracing the transmission of PZA-resistant Mycobacterium tuberculosis strains among patients. MATERIALS AND METHODS: The study included 71 PZA-resistant M. tuberculosis strains isolated from 62 Polish and 9 German patients. All strains were analysed using minimal inhibitory concentration value determination, pncA mutation analysis, spoligotyping, 24-loci mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) and insertion sequence (IS) 6110 restriction fragment length polymorphism (RFLP) fingerprinting. RESULTS: In 63 isolates, 37 (88.7%) different mutations in the pncA gene were observed, 13 of which had not been previously reported; 11 molecular families with the same MIRU-VNTR and IS6110-RFLP pattern were found. The same mutation was identified in three families, while different ones were identified in the remaining families. CONCLUSION: Mutations in the pncA gene are a major cause of PZA resistance in M. tuberculosis. pncA mutation analysis can be used to obtain valuable additional information, but should be applied with caution for epidemiological analysis.

Molecular analysis of drug-resistant Mycobacterium tuberculosis isolates collected in central Poland.

In total, 48 Mycobacterium tuberculosis clinical isolates from 48 patients with drug-resistant pulmonary tuberculosis (TB) in central Poland during 2004 were analysed by spoligotyping and IS6110-Mtb1/Mtb2 PCR. Twelve (25%) isolates were clustered by spoligotyping in combination with IS6110-Mtb1/Mtb2 PCR. Patients from whom these isolates were obtained were assumed to have developed TB as a result of recent transmission. Spoligotyping, used alone, overestimated the number of clustered isolates. However, combined use of spoligotyping and IS6110-Mtb1/Mtb2 PCR was an efficient approach for revealing clonal relatedness among M. tuberculosis isolates.

Humoral immune response against mycobacterial antigens in bronchoalveolar fluid from tuberculosis patients.

Resistance to tuberculosis (TB) is cell-mediated but a humoral response is common and may be correlated with the lack of effective local cellular defense mechanisms. The goal of the study was to evaluate IgG, IgA, and IgM-mediated humoral immune response against 38-kDa+16-kDa and 38-kDa+lipoarabinomannan (LAM) mycobacterial antigens in bronchoalveolar fluid (BALF) from patients with pulmonary TB. Non-tuberculosis (NTB) patients were used as control. 179 BALF samples (56 TB and 123 NTB) were examined. Commercially available ELISA-based assays against proteins 38-kDa and 16-kDa or 38-kDa plus LAM were used. Three different dilutions of BALF: 1:1; 1:10, and 1:50 (100) were tested. Only the results obtained with the 1:10 dilution allowed distinguishing TB and NTB groups. The mean IgG level for 38-Da+LAM was significantly higher in the TB than that in the NTB group (P<0.0001). The mean IgA level for 38-kDa+LAM also was higher in the TB group (P<0.05). No difference was observed between TB and NTB groups in the titer of IgM antibodies. These findings indicate that TB is associated with the presence of detectable levels of antibodies in BALF. The antibody response is highly heterogeneous. This phenomenon results from the balance between pathogen and host immune system. The tests examined for detection of IgG in BALF can be used in combination with other diagnostic methods to increase diagnostic accuracy of pulmonary TB.

Molecular characterisation of streptomycin-resistant Mycobacterium tuberculosis strains isolated in Poland.

Primary drug resistance of Mycobacterium tuberculosis strains in Poland increased two-fold between 1997 and 2000. Among 3705 drug-resistant strains investigated in 2000, 169 were resistant to streptomycin alone or in combination with isoniazid, rifampicin and/or ethambutol. The molecular basis of streptomycin resistance for 88 (52%) of these strains in comparison with 15 susceptible controls was determined. The most prevalent mutation was the single substitution Lys43Arg in the rpsL gene, found in 30.7% of the strains analysed. However, as many as 51% of the strains investigated carried no mutation in the rpsL or rrs genes. The multiple mutations present in two Beijing family strains were also identified.

Molecular epidemiology of drug-resistant Mycobacterium tuberculosis strains isolated from patients with pulmonary tuberculosis in Poland: a 1-year study.

OBJECTIVE: To characterise drug-resistant Mycobacterium tuberculosis strains isolated in Poland and to estimate the amount of recent transmission in the population. DESIGN: M. tuberculosis strains isolated from 251 patients with resistant pulmonary tuberculosis in Poland in 2000 were analysed by spoligotyping and IS6110 DNA fingerprinting. Part of the strains was also characterised by sequencing of the rpoB, katG and/or the regulatory region of the inhA gene. RESULTS: Using combined spoligotyping/IS6110-RFLP defined clusters, 29% of the strains were clustered, suggesting possible recent transmission. In some cases, transmission links among strains in clusters could be confirmed by epidemiological data and in addition, for most of the strains, by analysis of the mutations associated with resistance to rifampicin and/or isoniazid. Younger age, sex, immigration and history of previous treatment were not associated with clustering, whereas multidrug-resistant disease was more likely to cluster. Strains of the Beijing family could also be found in Poland, although with a much lower frequency than in the neighbouring countries. CONCLUSION: Transmission of drug-resistant M. tuberculosis strains was demonstrated, which might contribute to the emergence of drug-resistant tuberculosis in Poland.

Drug-resistant tuberculosis in Poland in 2000: second national survey and comparison with the 1997 survey.

SETTING: Three years after the last survey of drug-resistant tuberculosis in Poland, a nationwide survey was conducted by the National Tuberculosis Reference Laboratory in cooperation with all TB laboratories. OBJECTIVE: To determine the prevalence and patterns of primary and acquired drug resistance among Mycobacterium tuberculosis isolates recovered from tuberculosis patients in Poland in 2000 and to compare the results with an earlier survey carried out in 1997. DESIGN: In a prospective survey based on the questionnaires and strains of M. tuberculosis from all 16 regional centres participating in the study, 3705 questionnaires and cultures were obtained from 3037 new and 668 previously treated patients excreting TB bacilli during the 12-month period of 2000. Tests of resistance to isoniazid, rifampicin, streptomycin and ethambutol were performed using Löwenstein-Jensen and the Bactec system. RESULTS AND CONCLUSION: Compared with the previous survey in 1997, the current survey showed a twofold increase in tuberculosis resistance in new cases: any resistance was 3.6% in 1997 vs. 6.1% in 2000 (P < 0.001), multidrug resistance was 0.6% vs. 1.2% (P < 0.01), and no cases of four-drug resistance in 1997 vs. 15 cases in 2000. No statistical differences were observed in the rate of acquired resistance in both surveys.

[Direct detection of mycobacterium tuberculosis complex in clinical specimens by gen-probe amplified direct test]. / Wykrywanie pratków gruzlicy w materialach klinicznych metoda genetyczna Gen-Probe Amplified Direct Test.

The amplified Mycobacterium tuberculosis direct test (MTD) (Gen-Probe Inc., San Diego, CA, USA) is a rapid technique of nucleic acid amplification which can be used directly on processed clinical specimens. It is based on the enzymatic amplification of ribosomal RNA via DNA intermediates, with detection of amplified product by an acridinum-ester-labeled DNA probe. The purpose of this study was to evaluate the clinical utility of this test for diagnosis of tuberculosis by comparing the sensitivity and specificity of the test with acid-fast smear, mycobacterial culture and clinical evaluation. The study included 399 specimens from patients, suspected of Tb which were submitted to the microbiological laboratory of our Institute over a 12 months period. Compared with bacterioscopy, conventional culture on L-J and rapid systems of cultivation (Bactec-450 Tb, MB/Bact, Bactec 960 MGIT) MTD had a sensitivity 93.2% and specificity 98.5%. We conclude, that MTD test which is completed within 6-8 hours, when used rationally, mainly in conjunction with routine smear and culture is a useful, rapid diagnostic test for suspected tuberculosis patients.

Primary and acquired drug resistance in Polish tuberculosis patients: results of a study of the national drug resistance surveillance programme.

OBJECTIVE: To determine the prevalence and patterns of primary and acquired drug resistance among Mycobacterium tuberculosis isolates recovered from tuberculosis patients in Poland. DESIGN: In a prospective survey, M. tuberculosis strains were collected from 3970 tuberculosis patients (2976 newly diagnosed cases and 994 previously treated patients) bacteriologically confirmed by culture between November 1996 and October 1997. METHODS: Drug susceptibility testing to isoniazid (INH), streptomycin, ethambutol and rifampicin (RMP) was performed on Löwenstein-Jensen medium according to the proportion method and/or using the radiometric Bactec 460 TB system. RESULTS AND CONCLUSION: The male to female ratio was 2.61:1. The patients were aged between 6 and 82 years, with 86% of males and 77% of females aged over 35 years. Primary resistance to any drug was found in 3.6% of new patients; any INH resistance was 2.6%, any RMP resistance was 0.7%, and multidrug resistance (to INH and RMP [MDR]) was 0.6%. In previously treated cases, resistance to any drug was 17.0%, any INH resistance 14.1%, any RMP resistance 7.8%, and MDR 7.0%. Drug-resistant tuberculosis does not present a big problem in Poland; primary drug resistance has been monitored since 1960 with decreasing frequency, and rates remain at the same level as 20 years ago. Studies such as this should be conducted regularly to monitor drug resistance in Poland in order to effectively manage national tuberculosis control efforts.

[Value of the immunochromatographic assay for detecting IgG antibodies against 38 kDa mycobacterial antigen in diagnosis of tuberculosis]. / Przydatnosc testu immunochromatograficznego wykrywajacego przeciwciala IgG przeciwko antygenowi pratka 38 kDa w rozpoznawaniu gruzlicy.

Despite of a fast development in the techniques of rapid identification of mycobacteria by molecular genetic techniques, serodiagnosis may be of special values as non-expensive, easy to perform method. Several serodiagnostic tests, principally those using immunoenzymatic (ELISA) methodology are available. The goal of our study was to evaluate one step coloured immunochromatographic assay detecting IgG antibodies against antigen 38 kDa (Rapid Test TB). Our material consisted of 278 serum samples--tuberculosis (n = 155), healthy (n = 36), sarcoidosis (n = 50), lung cancer (n = 25) mycobacterial infections other than tuberculosis (n = 12). Tuberculosis group consisted of new culture positive cases (n = 66), new culture negative cases (n = 23), chronic cases (n = 43) and extrapulmonary TB (n = 23). Specificity of 96% and sensitivity of 54% was obtained. In pulmonary TB sensitivity of 50% and in extrapulmonary TB of 74% was obtained. In chronic cases sensitivity of 70% and in new cases of 40% was received. Sensitivity of 44% in new culture positive cases and 30% in new culture negative cases was obtained. We conclude that immunochromatographic test may be a very useful tool improving tuberculosis diagnosis, especially in extrapulmonary tuberculosis. Strip test may be an interesting alternative as it is an extremely simple, rapid, and cheap technique.

[Usefulness of measuring serum IGG antibodies against A60 mycobacterial antigen for diagnosis of tuberculosis]. / Przydatnosc oznaczania w surowicy przeciwcial IgG przeciwko antygenowi A60 w diagnostyce gruzlicy pluc.

Measurement of antimycobacterial antibody may be used as potential diagnostic tool in tuberculosis. The aim of the study was to evaluate the diagnostic value of serum IgG level against A60 mycobacterial antigen measured by ELISA method. Material consisted of 144 persons divided into 5 groups (76 tuberculosis patients, 20 sarcoidosis patients, 17 lung cancer patients, 8 patients with mycobacterial infections other than tuberculosis and 23 healthy controls). In the tuberculosis group there were 50 culture positive cases and 26 culture negative ones, 43 new cases and 32 chronic cases. Positive results were obtained in 51% of tuberculosis patients. Sensitivity increased to 62% in culture positive group and 63% in chronic cases. Specificity of the test was 96%. The results indicate that Immunozyme Mycobacterium test is a valuable tool in tuberculosis diagnosis.

[Results of examination in healthy volunteers of rifampicin and isoniazid bioavailability from Polish two-drug combination capsules of rifamazid used for tuberculosis treatment]. / Wyniki badania u zdrowych ochotników biodostepnosci ryfampicyny i izoniazydu z Polskiej kapsulki dwulekowej do leczenia gruzlicy rifamazid.

Purpose of the study was the examination of the relative bioavailability of rifampicin and isoniazid from Rifamazid (fixed-dose combination) and bioavailability of each drug in free simultaneous combination. The study was carried out as an open randomized cross-over study comprising 16 healthy volunteers for study of rifampicin and 15 healthy volunteers (slow acetylators) for isoniazid. The results have shown, that the pattern of absorption, elimination, plasma concentrations and pharmacokinetics parameters were found similar after administration of the drugs in free and in fixed combination.

Bioavailability of rifampicin, isoniazid and pyrazinamide from fixed-dose combination capsules.

SETTING: The absorption of rifampicin, isoniazid and pyrazinamide was tested after administration of each drug in free combinations and in a fixed-dose combination of the three drugs, known as Trifazid. OBJECTIVE: To examine the relative bioavailability of rifampicin, isoniazid and pyrazinamide after oral administration of the drugs given alone in comparison to that of the same drugs after administration of Trifazid. DESIGN: An open, randomized, cross-over study comprising 16 healthy volunteers. RESULTS: The pattern of absorption, plasma concentrations and pharmacokinetic parameters were very similar after administration of the drugs in free and fixed combinations. CONCLUSION: The triple combination of antituberculosis drugs could replace the separate drugs in the treatment of tuberculosis.

[A new, automatic, non-radiometric system for culturing MB/BACT bacilli and its value in the microbiologic diagnosis of tuberculosis]. / Nowy, automatyczny, nieizotopowy system hodowli pratków MB/BACT i jego wartosc w diagnostyce mikrobiologicznej gruzlicy.

The MB/BacT is a fully automated, rapid, non-radiometric system, for the culture of Mycobacteria for clinical samples other than blood. CO2 production is measured and reported as reflective units from the MB/BacT Process Bottle colorimetric sensor. We are evaluating the MB/BacT system in comparison to our routine culture method--culture on egg media (glycerol and puruvate), in the Bactec 460-Tb radiometric machine and in MB Redox. A total of 286 clinical samples from respiratory tract were inoculated into three or four culture systems and incubated at 37 degrees C. There was only little difference in mean time to detection between the MB/BacT and Bactec system (for M.tuberculosis 11 days versus 9 days), but both systems were faster than egg media (16 days). Contamination rates for MB/BacT system was 8.4%, and for Bactec 6.9%. The colorimetric technology used in the MB/BacT is a rapid and sensitive for the measurement of mycobacterial growth.

[Usefulness of redux properties in acid-resistant bacilli for rapid detection of their growth in culture]. / Wykorzystanie zdolnosci reduckcyjnych pratków kwasoopornych do szybkiego wykrywania ich wzrostu w hodowlach.

MB REDOX a new system for detection of Mycobacterium is described. It is a modified, serum supplemented Kirchner-medium containing a colourless tetrazolium salt reduced by the redox system used for self-indicating microbial growth. In this study we have tested sensitivity and time of isolation of Mycobacteria in MB REDOX and compared to the other different systems of cultivation.