Introduction: Pyrazinamide (PZA) is an important first-line anti-tuberculous drug, which is applied together with INH, RMP, EMB, and SM. This drug plays a unique role in the first phase of TB therapy because it is active within macrophages and kills tubercle bacilli. Testing the resistibility of Mycobacterium tuberculosis to PZA is technically difficult because PZA is active only at acid pHs. Therefore, routine drug resistibility testing of M. tuberculosis for PZA is not performed in many laboratories. The objective of our study was to estimate the resistibility for PZA among M. tuberculosis isolates from polish patients in the years 2000–2008. Material and methods: We analyzed M. tuberculosis strains with different resistibility to first-line anti-tuberculous drugs. The strains were isolated from 1909 patients with tuberculosis. The strains were examined for PZA resistibility by the radiometric Bactec 460-TB method. The PZA-resistant strains were examined for the following MIC PZA for drug concentrations: 100, 300, 600, 900 μg/mL. Results: PZA resistance among M. tuberculosis strains was found in 6.7% of untreated patients and in 22.2% of previously treated patients (p < 0.001). In both groups, resistance to PZA correlated with drug resistance for INH+RMP+SM+EMB—in 32.7% of untreated patients and in 34.5% previously treated patients (p < 0.8). PZA-monoresistant strains were observed in 20.8% of untreated patient groups. Among the resistant strains: in 3.4% MIC for PZA was > 100 μg/mL, in 11.6% ≥ 300 μg/mL, in 8.9% ≥ 600 μg/mL, and in 76% ≥ 900 μg /mL. Conclusions: Among M. tuberculosis strains, PZA resistance was found in 6.7% of untreated patients and in 22.2% of previously treated patients. Among the PZA-resistant strains, very high MIC values for PZA (≥900 μg/mL) were revealed for 76% M. tuberculosis strains.
Abstract Introduction Since 2009, Poland has been recognised as a country officially free of bovine tuberculosis (bTB), although in each year of the last five there were from 8 to 18 outbreaks of the disease. In 2008–2016, the largest number of cattle infected with bovine mycobacteria were eliminated in the Masovian Province (the central region of Poland) and the largest number of outbreaks of this zoonosis were recorded in this area. The close proximity of farms where bTB was found led to the suspicion that tuberculosis could have been transmitted between the affected herds. The aim of the study was the molecular characterisation of the pertinent M. bovis / caprae strains and determination of the epidemiological relationship of various bTB outbreaks. Material and Methods The material for microbiological tests came from 119 cattle ( Bos taurus ) from nine herds located in five provinces, neighbouring the Masovian Province. Results Laboratory tests of tissue material gave results confirming tuberculosis in 54 (45%) animals. All strains belonged to the Mycobacterium bovis species. A two-step analysis of genetic affinity allowed 50 strains to be identified as phylogenetically closely related and separated between three genetic clusters consisting of 2 to 27 strains. Conclusion Based on the results of genotyping, bTB outbreaks were found in three herds, and three transmission chains were identified among these herds.
Pyrazinamide (PZA) is an important first-line anti-tuberculosis drug that is generally administered with isoniazid, rifampicin, ethambutol and streptomycin.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.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.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.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.
Non-tuberculous mycobacterial lung disease (NTMLD) is diagnosed in 3-6% of patients (pts) with predisposing chronic lung diseases such as COPD and cystic fibrosis. We reported previously the occurrence of NTMLD due to M. kansasii in 6/156 CTEPH pts (3.8%) treated in our centre from 2002 to 2008. The aim of the present study was to analyse the possible causes of NTMLD development in CTEPH pts. Methods: Clinical data, chest CT scans and hemodynamic variables of 156 CTEPH pts were analysed. Reconstructions were obtained to allow for better visualisation of lung density on chest CT scan. Results: Marked mosaic perfusion of grade 3-4 was significantly more frequent on CT scans in CTEPH pts who developed NTMLD in comparison to the others (100% vs 64%, p=0.024). Moreover NTMLD, described as focal consolidations with cavitation and satelite micronodules, developed in the areas of lung hypoperfusion. Pulmonary artery diameter on chest CT scan was 40 (36-42) mm in NTMLD pts comparing to 35 (26 -62) mm in the remaining ones (p=0.015). CTEPH patients with NTMLD were slimmer than the others: BMI were 23.4 (17.6-27.7) kg/m2 and 28.45 (15.6-55.1) kg/m2 respectively, p=0.039. NTMLD pts had also lower cardiac indexes and lower mixed venous blood oxygen saturation in comparison to the group without mycobacteriosis. Conclusions: Vascular pathology resulting in the areas of hypoperfusion within the lungs is probably the reason of growing susceptibility to NTMLD in CTEPH. This phenomenon might be responsible for local defence disturbances. The disease occurrence in younger and slimmer group of CTEPH pts is also observed.
The aim of the present study was to determine the susceptibility of budgerigars (Melopsittacus undulatus) to infections with different Mycobacterium species. For inoculations the following Mycobacterium species were used: Mycobacterium avium subsp. avium, Mycobacterium bovis subsp. bovis, Mycobacterium tuberculosis subsp. tuberculosis, Mycobacterium intracellulare and M. fortuitum subsp. fortuitum. The bacterial suspension was administrated intramuscularly and all the birds were monitored for 70 days starting from the day of inoculation. During the experiment clinical examination, X-ray scans, plate agglutination tests, tuberculin tests, faeces smear preparations and culture of mycobacteria were performed. The study showed that M. bovis subsp. bovis was the most pathogenic Mycobacterium species for budgerigars. After inoculation, the bacilli induced tuberculosis-typical, clinical signs and necropsy findings. In two out of six birds infected with M. bovis subsp. bovis radiological changes were also visible. Birds inoculated with other Mycobacterium species did not show any typical symptoms of infection, and only the results of histopathological and bacteriological examinations indicated the presence of infection.
Mycobacteriosis caused by Mycobacterium avium subsp. avium was observed in a parental loft of 70 meat-breed pigeons. It was decided to undertake treatment as the birds represented a substantial value to the owner. A multiagent therapy using azithromycin, marbofloxacin, and ethambutol was administered. After 4 mo of therapy, the desired results were not obtained. At the end of treatment, the birds were in poor general condition, with white blood cells above 20 g/L, and after clutching, 2-yr-old and older birds were euthanatized. Overall, postmortem lesions were found in 17 out of 49 necropsied individuals. Slide agglutination tests with a M. avium subsp. avium lysate were conducted in all examined pigeons. In 28 pigeons, blood count was conducted once a month during therapy, while in 24 pigeons, a tuberculin sensitivity test was conducted before the planned euthanatization. The tuberculin sensitivity test did not prove useful in the diagnosis of ill individuals. Slide agglutination yielded positive results in only four birds, all of which also had postmortem lesions. Blood count in a large number of cases allowed distinguishing between ill and healthy individuals, which was used for subsequent selection. The comparison of cultured strains with the (CCG)4-based PCR method showed the variation of M. avium isolates up to a maximum of 30%. The described case proves that the treatment of mycobacteriosis in pigeon flocks is not effective, mainly due to the high resistance to M. avium subsp. avium. In addition, therapy may contribute to an even greater increase in mycobacterial resistance to antibiotics, which may pose a potential risk to public health.
ABSTRACT We demonstrated that not whole Mycobacterium tuberculosis but its particular antigens, hsp70 Mtb , hsp65 Mtb , and hsp16 Mtb , are present in lymph node tissues of patients with sarcoidosis (SA). hsp16 Mtb occurs in the early stage of SA, whereas hsp70 Mtb occurs in stage II of SA. hsp65 Mtb is highly expressed in the capillary vessels in lymph node tissues in patients with SA.
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