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.

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.

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.

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.

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.

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.

[Pulmonary infections in patients with lung cancer during antineoplastic therapy]. / Zakazenia u chorych na raka pluca w czasie leczenia przeciwnowotworowego.

162 pulmonary infection episodes were observed in 94 patients with lung cancer undergoing antineoplastic therapy. 80 (40%) episodes occurred during leukopenia. Elevation of leucocyte count was seen in 12 episodes only. Elevated body temperature was the only sign in 20 episodes, of which in 7 cases microorganisms were cultured from the blood. Purulent pulmonary infections were observed in 71 episodes, in 66 the causative agent was identified. Purulent urinary tract infections were observed in 29 episodes, of which in 28 the microorganisms were identified. A coexistent pulmonary and urinary tract infection was seen in 13 cases, of which in all the causative agent was identified. Purulent infections of the nasopharyngeal mucosal membranes were observed in 16 cases, while herpes zoster in 13. The most often isolated organism in these cases were: Gram negative rods (E. coli, Klebsiella sp., Proteus sp., Hemophilus influenzae); less commonly Gram positive bacteria were isolated, mainly Staphylococcus aureus. Candida sp. was the most common fungus that was isolated from these patients. In four cases Candida was isolated from blood.

[Influence of food on the bioavailability of isoniazid in health volunteers]. / Wplyw pozywienia na biologiczna dostepnosc izoniazydu (INH) u zdrowych ochotników.

From the theoretical point of view, food intake exerts a complex influence on the bioavailability of drugs. It may interfere with tablet disintegration, drug dissolution and drug transit through the gastrointestinal tract, may also affect the metabolic transformation of drugs in the gastrointestinal wall and the liver. The knowledge about the influence of food is very important in treatment of tuberculosis, where the drugs are taken once daily or even less frequently. We studied the isoniazid concentrations in blood serum of 20 volunteers (12 slow and 8 fast acetylators of INH) both sexes, after the drug intake 2 hours before breakfast and immediately after breakfast. Concentrations of INH were tested by biological method with Mycobacterium aurum REB as a standard strain. The results have shown, that Cmax (3.55 +/- 0.29 versus 2.56 +/- 0.49 in fast acetylators and 4.34 +/- 0.2 versus 3.67 +/- 0.37 in slow acetylators), tmax (0.75 +/- 0.26 versus 1.31 +/- 0.37 in fast acetylators and 0.87 +/- 0.31 versus 1.79 +/- 0.62 in slow acetylators and AUC0-t (6.6 +/- 1.34 versus 5.47 +/- 1.5 in fast acetylators and 21.82 +/- 2.57 versus 18.61 +/- 2.57 in slow acetylators) are significantly decreased after ingestion of INH with breakfast.

[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.

[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.

[Drug sensitivity of bacillus strains M. avium-intracellulare (MAIC), M. kansasii cultured from patients with mycobacteriosis before treatment]. / Wrazliwosc na leki szczepów pratków M. avium-intracellulare (MAIC), M. xenopi i M. kansasii, wyhodowanych od chorych z mykobakteriozami przed rozpoczeciem leczenia.

The aim of study was to evaluate a drug sensitivity of M. avium-intracellulare, M. xenopi and M. kansasii cultured from 55 patients with mycobacterioses. The identification of strains was performed with morphological and biochemical tests and thin-layer chromatography. Resistance tests were done on egg L-J and agar media for selected drugs. It was documented that MAIC strains were non-sensitive on isoniazid and rifampicin, and other ones. The most active drug was cycloserine inhibiting growth of 80% but rifabutine--50% of strains. M. xenopi strains were sensitive for tested drugs including isoniazid and rifampicin (about 30%). M. kansasii strains were in 100% sensitive for cyclosporine, rifabutine, Davercin and ofloxacin and partly for isoniazid, streptomycin and Augmentin.

[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.

[The role of bronchoscopy and bronchoalveolar lavage in diagnosis of pulmonary tuberculosis in AIDS]. / Znaczenie bronchoskopii i plukania oskrzelowo-pecherzykowego w rozpoznawaniu gruzlicy pluc u chorych na AIDS.

Bronchoscopy was carried out in 32 HIV seropositive patients, most with AIDS during the period between January 1992 and August 1993. In 14 patients tuberculosis was diagnosed, in 13 it was bacteriologically confirmed. The mean age of the examined patients was 35.5 years (range 22-49 years). In 50% of the BAL samples bacterioscopy was positive. Bacteriological examination of the sputum and BAL fluid (bacterioscopy and culture) produced a confirmation of tuberculosis in 99.9% of the cases.

[Diagnostic value of IgG antibody levels against 38 kDa mycobacterial antigen]. / Wartosc diagnostyczna poziomu przeciwcial IgG przeciwko antygenowi 38 kDa pratka gruzlicy.

Tuberculosis diagnosis bases on clinical and radiological symptoms and identification of mycobacteria. Accuracy of both methods is limited. Therefore reliable serological test would have considerable advantage. The present study was aimed at evaluating IgG-mediated immune response against specific mycobacterial antigens 38 kDa in group of 200 patients and control subjects. Our material consisted of 104 tuberculosis patients, 25 with sarcoidosis, 24 with lung cancer, 13 with bacterial or fungal pulmonary infection, 8 with mycobacterial infections other than tuberculosis and 26 healthy persons. We used commercially available ELISA based kits (Pathozyme TB-complex). Specificity of 100% and sensitivity of 49% was achieved. Sensitivity increased to 59% in chronic cases and to 52% in culture positive cases. Sensitivity decreased to only 14% in group of new culture negative cases. Measurement of IgG serum level against 38 kDa can be helpful in tuberculosis diagnosis. As the test lacks falsely positive results it indicates its high positive predictive value.