Investigation of species distribution of nontuberculosis mycobacteria isolated from sputum samples in patients with suspected pulmonary tuberculosis.

Aims: Rapid and accurate identification of mycobacteria is important for the species-specific treatment of the disease. The aim of this study was the identification at the species level of 34 nontuberculous mycobacteria strains isolated from respiratory tract samples and 14 reference strains as by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Materials and Methods: Isolates derived from clinical specimens were subcultured in the Lowenstein-Jensen medium. Deoxyribonucleic acid isolation was carried out using the boiling method. PCR amplification was performed using primers specific to the hsp65 gene region. The PCR products were digested BstEII and HaEIII enzymes. All samples were studied comparatively by two different centers. Results: In our study, the most common species were found to be Mycobacterium intracellulare in 23.52% (8/34). The performance of the PCR-RFLP method in detecting mycobacteria was found to be 82.35%. Conclusions: The PCR-RFLP method is a rapid, cheap, and practical method for the identification of mycobacteria.

Spoligotyping and polymerase chain reaction based mycobacterium bovis strains typing with methods (enterobacterial repetitive intergenic consensus-polymerase chain reaction, randomly amplified polymorphic dnas-polymerase chain reaction and out polymerase chain reaction).

Background: In this study, it was aimed to investigate Mycobacterium bovis strains isolated from lungs and lymph nodes of slaughtered animals on clonal level by using different methods such as spoligotyping, enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR), randomly amplified polymorphic DNAs (RAPD-PCR) and OUT-PCR. Comparative evaluation of these methods was further conducted. Methods: A total of 38 M. bovis isolates were evaluated in the study. DNA isolation of all M. bovis strains isolated from pruvat free Löwenstein Jensen medium was done by boiling method for ERIC-PCR, RAPD-PCR, and OUT PCR. Mickle device was used for DNA isolation for spoligotyping method. Results: In 38 M. bovis isolates examined in our study, 4 different groups were determined by spoligotyping and RAPD-PCR test methods, and 5 different groups were detected in ERIC-PCR tests. In the OUT-PCR tests, the band which provides sufficient type separation was not observed. Conclusion: ERIC-PCR, RAPD-PCR, and OUT-PCR methods are easily applicable, simple, and relatively inexpensive methods for evaluating the differences between origins in the typing of M. bovis. The tests need to be evaluated in more detail with extensive studies.

Investigation of Bartonella henselae in cats in Ankara, Turkey.

The purpose of this study was to determine Bartonella henselae prevalance in cats in Ankara. Whole bloods and sera collected from 256 cats were investigated for the presence feline Bartonella species by culture and sera were tested for the presence of antibodies against B. henselae IgG using immunofluorescence assay. Bartonella species were isolated by blood culture from 24 (9.4%) cats. Bartonella isolates were subjected to restriction fragment length polymorphism (RFLP) by using TaqI and HhaI endonucleases to identify species. Twenty-one isolates were determined as B. henselae and three of 24 isolates were determined as Bartonella clarridgeiae with RFLP. The bacteraemia prevalence and seroprevalence of B. henselae IgG antibodies in cats was detected as 8.2% and 18.6% respectively. This is the first report on B. henselea and B. clarridgeiae in cats in Turkey.

Comparison of the tuberculin skin test and the quantiferon test for latent Mycobacterium tuberculosis infections in health care workers in Turkey.

The aim of this study was to compare the efficacy of the tuberculin skin test (TST) and the quantiferon test (QFT) for detecting latent tuberculosis infection (LTBI) in health care workers (HCWs). Seventy-six participants who were working in Duzce University Hospital, where tuberculosis patients were being treated, were included in the study. TST was performed according to the Mantoux technique. QFT was performed in accordance with the manufacturer's instructions. A positive TST result was defined as an induration diameter of > or = 15 mm. TSTs were positive in 41 of 76 participants (53.9%) and QFT was positive in 65 of 76 participants (85.5%). There was a significant difference between the numbers of QFT-positive and TST-positive cases (P=0.02). When the induration diameter of TST was > or = 20 mm, QFT positivity was 100%. Multivariate analysis revealed that there was a significant correlation between the percentage of patients with QFT positivity and the induration diameter of TST (P=0.009). QFT thus seems to be more effective for LTBI diagnosis than TST. However, large-scale trials including quantitative measurement of QFT in subgroups taking into account the division where HCWs are employed and the different results of TST might clarify the usefulness of QFT in LTBI diagnosis.

Modulating effects of mucoregulating drugs on the attachment of Haemophilus influenzae.

Non-typable Haemophilus influenzae (NTHI) is one of the three major pathogens implicated in human respiratory infections. The ability to attach with pharyngeal epithelial cells is an important factor for infection and virulence. In the present study we describe the effects of two mucoregulating drugs, S-carboxymethylcysteine (S-CMC) and ambroxol, on the attachment of NTHI to pharyngeal epithelial cells. There was a significant (P < 0.0001, < 0.001 and <0.01) decrease of attachment (8.8 +/ 2.4, 9.2+/-2.5 and 15.4 +/- 5.7 bactreria/cell) compared with the control (17.5 +/- 2.9, 15.5 +/- 3.1 and 18.8 +/- 6.8 bacteria/cell) after cells were treated wth S-CMC at a dose of 100, 10 and 1 microg/ml. After attachment assay, cells treated with S-CMC (100 microg/ml) showed a significant decrease (P < 0.01) of attached bacteria (3.1 +/- 0.8 bacteria/cell) compared with the control (5.9 +/- 1.8 bacteria/cell). Treatment of cells with ambroxol did not influence bacterial attachment. By scanning electron microscopic observation it was found that NTHI attaches to the surface elevations (microplicae) of human pharyngeal epithelial cells. Atomic force microscopic observation revealed that the surface potential of microplicae decreased significantly in cells treated with S-CMC compared with the untreated control cells. As bacteria with negative surface charge attach to the positively charged domain, i.e. microplicae of human pharyngeal epithelial cells, this study suggests that the decrease of attachment of NTHI with epithelial cells after treatment with S-CMC was possibly due to the decrease of surface charge. This study suggests that S-CMC decreases the episodes of respiratory infections in patients with respiratory diseases both by inhibiting the attachment of bacteria to the upper respiratory tract, and by detaching the adherent one.