Genotyping of Mycobacterium tuberculosis clinical isolates in two cities of Turkey: Description of a new family of genotypes that is phylogeographically specific for Asia Minor

BACKGROUND: Population-based bacterial genetics using repeated DNA loci is an efficient approach to study the biodiversity and phylogeographical structure of human pathogens, such as Mycobacterium tuberculosis, the agent of tuberculosis. Indeed large genetic diversity databases are available for this pathogen and are regularly updated. No population-based polymorphism data were yet available for M. tuberculosis in Turkey, at the crossroads of Eurasia. RESULTS: A total of 245 DNAs from Mycobacterium tuberculosis clinical isolates from tuberculosis patients residing in Turkey (Malatya n = 147 or Ankara n = 98) were genotyped by spoligotyping, a high-throughput genotyping method based on the polymorphism of the Direct Repeat locus. Thirty-three spoligotyping-defined clusters including 206 patients and 39 unique patterns were found. The ST41 cluster, as designated according to the international SpolDB3 database project, represented one fourth and when gathered to three genotypes, ST53, ST50 and ST284, one half of all the isolates. Out of 34 clinical isolates harboring ST41 which were further genotyped by IS6110 and by MIRU-VNTR typing, a typical 2-copy IS6110-RFLP pattern and a "215125113322" MIRU-VNTR pattern were observed among 21 clinical isolates. Further search in various databases confirms the likely Turkish-phylogeographical specificity of this clonal complex. CONCLUSION: We described a new phylogeographically-specific clone of M. tuberculosis, designated LAM7-TUR. Further investigations to assess its frequency within all regions of Turkey and its phylogeographical origin and phylogenetic position within the global M. tuberculosis phylogenetic tree will shed new light on its endemicity in Asia Minor

Demographic and microbial characteristics of extrapulmonary tuberculosis cases diagnosed in Malatya, Turkey, 2001-2007

<p>Abstract</p> <p>Background</p> <p>Extrapulmonary tuberculosis (EPTB) has an increasing rate in Turkey. The reason remains largely unknown. A better understanding of the demographic and microbial characteristics of EPTB in the Turkish population would extend the knowledgebase of EPTB and allow us to develop better strategies to control tuberculosis (TB).</p> <p><b>Methods</b></p> <p>We retrospectively evaluated clinical and laboratory data of 397 bacteriologically-confirmed TB cases diagnosed during an eight year-period using by chi-square analysis and multivariate logistic regression model.</p> <p>Results</p> <p>Of the 397 study patients, 103 (25.9%) had EPTB and 294 (74.1%) had pulmonary tuberculosis (PTB). The most commonly seen two types of EPTB were genitourinary TB (27.2%) and meningeal TB (19.4%). TB in bone/joints, pleural cavity, lymph nodes, skin, and peritoneal cavity occurred at a frequency ranging from 9.7% to 10.7%. The age distribution was significantly different (P < 0.01) between PTB and EPTB, with patients older than 45 years tending to have an increased risk of EPTB. Furthermore, the distribution of different types of EPTB differed significantly among age groups (P = 0.03). Meningeal and bone and/or joint TB were more commonly observed among the male patients, while lymphatic, genitourinary, and peritoneal TB cases were more frequently seen among females. Unique strain infection was statistically significantly associated with EPTB (OR: 2.82, 95% CI [1.59, 5.00])</p> <p>Conclusions</p> <p>EPTB accounted for a significant proportion of TB cases in Malatya, Turkey between 2001 and 2007. The current study has provided an insight into the dynamics of EPTB in Malatya, Turkey. However, the risk factors for having EPTB in Malatya, Turkey remain to be assessed in future studies using population-based or randomly selected sample.</p

Distribution of Spoligotyping Defined Genotypic Lineages among Drug-Resistant Mycobacterium tuberculosis Complex Clinical Isolates in Ankara, Turkey

Background: Investigation of genetic heterogeneity and spoligotype-defined lineages of drug-resistant Mycobacterium tuberculosis clinical isolates collected during a three-year period in two university hospitals and National Tuberculosis Reference and Research Laboratory in Ankara, Turkey. Methods and Findings: A total of 95 drug-resistant M. tuberculosis isolates collected from three different centers were included in this study. Susceptibility testing of the isolates to four major antituberculous drugs was performed using proportion method on Löwenstein–Jensen medium and BACTEC 460-TB system. All clinical isolates were typed by using spoligotyping and IS6110-restriction fragment length polymorphism (RFLP) methods. Seventy-three of the 95 (76.8%) drug resistant M. tuberculosis isolates were isoniazid-resistant, 45 (47.4%) were rifampicin-resistant, 32 (33.7%) were streptomycinresistant and 31 (32.6%) were ethambutol-resistant. The proportion of multidrug-resistant isolates (MDR) was 42.1%. By using spoligotyping, 35 distinct patterns were observed; 75 clinical isolates were grouped in 15 clusters (clustering rate of 79%) and 20 isolates displayed unique patterns. Five of these 20 unique patterns corresponded to orphan patterns in th

The investigation of the effect of NAD, H2O2 and weak magnetic field on the antibacterial mechanism of isoniazid (INH) that first line antibiotic against M. tuberculosis agent

Tuberculosis is an infectious disease, which is caused by the Mycobacterium tuberculosis complex. This disease leads to up to 1.3 million deaths out of more than eight million cases every year. A prodrug called isoniazid has been proven to be effective and widely used in the treatment of infections caused by tuberculosis. Despite its use for more than six decades clinically, the action mechanism of this prodrug is yet to be elucidated. INH action agains mycobacteria requires catalase&#8722;peroxidase (KatG) function, and IN-NAD adduct formation is catalyzed in vitro by M. tuberculosis KatG under a variety of conditions. Low-intensity EMF (Electromagnetic Field) has been used in the&#8322;rapeutic practices in addition to its use in telecommunication systems and food protection. EMF is used in medicine and food industries especially for its bactericidal effects. In this study, we aimed to investigate the effects of weak magnetic field application and the addition of NAD and H&#8322;O&#8322; on the action mechanism of isoniazid. We added H&#8322;O&#8322; and NAD individually and together, to the different groups at varying concentrations. Also, one experimental group was exposed to a 5mT, 50Hz magnetic field for 4 to 5 hours per day (total of 45 hours in 10 days). The agar proportion method was used to evaluate the results. It was determined that the addition of 100 &#956;M NAD and H&#8322;O&#8322; together increased the effectiveness of isoniazid to some extent. However, the application of a weak magnetic field did not change the effectiveness of the drug. [Med-Science 2020; 9(1.000): 207-11

Integration of Mycobacterium tuberculosis Drug Susceptibility Testing and Genotyping with Epidemiological Data Analysis To Gain Insight into the Epidemiology of Drug-Resistant Tuberculosis in Malatya, Turkey▿

Drug-resistant tuberculosis (TB) presents a major challenge to global TB control. To gain a better understanding of drug-resistant TB epidemiology in Malatya, Turkey, we conducted the present study using 397 Mycobacterium tuberculosis clinical isolates collected from Malatya, Turkey, in recent years (2000-2007). Resistance to any anti-TB drug was found in 29% (114 of 397) of the study isolates, while the multidrug resistance (MDR) rate was ∼4.5% (18 of 397). Resistances to isoniazid (15.5%) and streptomycin (13.4%) were about twice as high as resistance to rifampin (RMP) (6.3%) and ethambutol (EMB) (6.0%). Importantly, 28% (7 of 25) of the RMP-resistant isolates were non-MDR isolates, as when a significant proportion of RMP-resistant isolates in a population are non-MDR, the predictive value of molecular detection of RMP resistance for MDR can be significantly reduced. Both identical and varied drug resistance patterns were seen in the same genotyping-defined clusters, suggesting that both primary and acquired resistance have contributed to the drug-resistant TB epidemic in Malatya, Turkey. In addition, drug-resistant cases were found to be more likely to be males (odds ratio [95% confidence interval], 1.82 [1.13, 2.94]), suggesting a potential role of gender in the epidemiology of drug-resistant TB in the study population. This study demonstrates that the integration of drug susceptibility testing with genotyping and epidemiological data analysis represents a useful approach to studying the epidemiology of drug-resistant TB

Synthesis of some heterocyclic phosphonates and their antibacterial and antifungal activities

197-200Fifteen heterocyclic phosphonate derivatives and their starting compounds have been evaluated in vitro for antibacterial and antifungal activities against standard strains: Enterococcus faecalis (ATCC 29212), Staphylococcus aureus (ATCC 2921)), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853) and yeast-like fungi ; Candida albicans and <span style="font-size:16.0pt; mso-bidi-font-size:10.0pt;font-family:" times="" new="" roman";mso-fareast-font-family:="" "times="" roman";mso-ansi-language:en-us;mso-fareast-language:en-us;="" mso-bidi-language:ar-sa"="">Candida tropicalis. <span style="font-size: 15.0pt;mso-bidi-font-size:9.0pt;font-family:" times="" new="" roman";mso-fareast-font-family:="" "times="" roman";mso-ansi-language:en-us;mso-fareast-language:en-us;="" mso-bidi-language:ar-sa"="">The compounds newly synthesised have been identified by 1H-NMR,FT-IR and micro analysis. Among the tested compounds 12, 13, 16, 17, 18, 22 and 23 are found effective to inhibit the growth of <span style="font-size:16.0pt;mso-bidi-font-size:10.0pt; font-family:" times="" new="" roman";mso-fareast-font-family:"times="" roman";="" mso-ansi-language:en-us;mso-fareast-language:en-us;mso-bidi-language:ar-sa"="">Candida albicans <span style="font-size:15.0pt;mso-bidi-font-size:9.0pt; font-family:" times="" new="" roman";mso-fareast-font-family:"times="" roman";="" mso-ansi-language:en-us;mso-fareast-language:en-us;mso-bidi-language:ar-sa"="">and <span style="font-size:16.0pt;mso-bidi-font-size:10.0pt;font-family: " times="" new="" roman";mso-fareast-font-family:"times="" roman";mso-ansi-language:="" en-us;mso-fareast-language:en-us;mso-bidi-language:ar-sa"="">Candida tropicalis at the MICs between <span style="font-size:14.5pt;mso-bidi-font-size:8.5pt;font-family:Arial;mso-fareast-font-family: " times="" new="" roman";mso-ansi-language:en-us;mso-fareast-language:en-us;="" mso-bidi-language:ar-sa;mso-bidi-font-style:italic"="">100-800 μg/mL. The compounds tested here generally do not exhibit considerable antibacterial activity at the concentration studied (100-800 <span style="font-size:14.5pt;mso-bidi-font-size:8.5pt; font-family:Arial;mso-fareast-font-family:" times="" new="" roman";mso-ansi-language:="" en-us;mso-fareast-language:en-us;mso-bidi-language:ar-sa;mso-bidi-font-style:="" italic"="">μg/mL), <span style="font-size:15.0pt;mso-bidi-font-size: 9.0pt;font-family:" times="" new="" roman";mso-fareast-font-family:"times="" roman";="" mso-ansi-language:en-us;mso-fareast-language:en-us;mso-bidi-language:ar-sa"="">except compound 18 which exhibits antibacterial activity against gram-positive bacteria at the MIC of 400 <span style="font-size:14.5pt;mso-bidi-font-size:8.5pt;font-family:Arial;mso-fareast-font-family: " times="" new="" roman";mso-ansi-language:en-us;mso-fareast-language:en-us;="" mso-bidi-language:ar-sa;mso-bidi-font-style:italic"="">μg/mL.</span

Synthesis and antimicrobial properties of cycloheptyl substituted benzimidazolium salts and their silver(I) carbene complexes

WOS: 000389640000008Due to increasing infections caused by microbes, there is an urgent need for the development of new effective antimicrobial agents. Silver-N-heterocyclic carbene (silver-NHC) complexes are a new class of antimicrobial agents. In this study, we aimed to synthesize highly lipophilic silver-NHC complexes. Four new complexes were synthesized by the reaction of the corresponding benzimidazolium salts and Ag2O in dichloromethane at room temperature. The synthesized compounds were characterized by H-1 NMR, C-13 NMR, IR and elemental analysis. The antimicrobial performances of benzimidazolium salts and silver complexes were tested against the standard bacterial strains Enterococcus faecalis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and the fungi Candida albicans and Candida tropicalis. Minimum inhibitory concentrations (MICs) of all compounds were determined. The obtained data demonstrate that all benzimidazolium salts and silver complexes inhibit the growth of bacteria and fungi. Silver complexes are more active than the corresponding benzimidazolium salts (MIC: 6.25 mu g/mL for Gram-positive bacteria and fungi)

Comparison of keypads and touch-screen mobile phones/devices as potential risk for microbial contamination

Conclusions: We found no significant difference between TMP/Ds and KMPs in terms of microbial contamination, but TMP/Ds harboured more colonies and total microbial counts increased with screen size

Synthesis, antioxidant and anti-microbial properties of two organoselenium compounds

WOS: 000351657400030PubMed ID: 25730792The aim of this study is synthesis of two different series of organoselenium compounds and available in vitro antioxidant and antimicrobial properties of thee synthetic compounds. The synthetic compounds were identified by H-1-NMR (300 MHz), C-13-NMR (75.5 MHz), FT-IR spectroscopic techniques and micro analysis. Antioxidant properties of two synthetic organoselenium compounds were determined by 1,1- dipheny1-2-picrylhydrazyl (DPPH) radical method, reducing power assay and beta-carotene bleaching method as in vitro. Antimicrobial effects of samples were assessed by the agar dilution procedure and using gram positive and gram-negative bacteria and yeast strains. Although 1,3-di-pmethoxybenzylpyrimidine-2-selenone showed better antiradical activity in DPPH test and higher protective activity on beta-carotene, 1-isopropyl-3-methylbenzimidazole-2-selenone was found to be better in reducing power and antimicrobial activity.Technological and Scientific Research Council of Turkey TUBITAK [TBAG-2259 (102T185)]We thank to Technological and Scientific Research Council of Turkey TUBITAK TBAG-2259 (102T185) for financial support of this work

Population-Based Molecular Epidemiological Study of Tuberculosis in Malatya, Turkey▿

This investigation describes drug resistance patterns and genotyping data on a total of 145 Mycobacterium tuberculosis strains isolated between 2000 and 2004 in Malatya, Turkey. Drug susceptibility results indicated a total of 20% resistant and 4.8% of multidrug-resistant isolates. Spoligotyping resulted in 25 unique patterns and 120 strains in 19 clusters (2 to 33 strains per cluster). When the results were compared to an international spoligotyping database, 19 of 25 unique patterns matched existing shared spoligotype international types (SITs). This led to the description of 38 SITs with 139 strains and 6 orphan patterns (not previously reported). Five of the SITs (SIT759, SIT1936, SIT1937, SIT1938, and SIT2285) were newly created. The most prevalent spoligotype was SIT41 (LAM7-TUR) with 33 (23.9%) isolates. The repartition of strains according to major M. tuberculosis clades (in decreasing order) was as follows: ill-defined T clade (45.7%) > Latin American and Mediterranean (LAM; 29%) > Haarlem (15.9%). Strains belonging to Central Asian (CAS), East-African Indian (EAI), Beijing, and Africanum clades were absent in this setting. IS6110-restriction fragment length polymorphism (RFLP) resulted in 19 clusters (52 strains), with a final clustering rate of 35.9% and a recent transmission rate of 22.8%. Typing based on mycobacterial interspersed repetitive units (MIRUs) permitted us to identify 65 patterns (23 orphan patterns and 42 patterns that matched existing MIRU international types in an updated database). The combination of the three typing methods allowed us to calculate a final clustering rate of 22% and a significantly lower transmission rate of 13.1%. The discrimination achieved by IS6110-RFLP/MIRUs was not significantly improved by adding spoligotyping results (1.4%). We conclude that our patient population is infected by diverse M. tuberculosis populations; however, the majority of the ongoing transmission is due to “evolutionary recent” tuberculosis lineages belonging to principal genetic group 2 (PGG2; Haarlem and LAM) and PGG3 (ill-defined T clade), and most of it is attributable to the LAM7-TUR sublineage with an enhanced phylogeographical specificity for Turkey. An absence of lineages belonging to PGG1 clones (EAI, CAS, and Beijing, essentially found in Central, South, and Southeast Asia), is noteworthy