Djursun Karasartova

Expression analysis of efflux pump genes among drug-susceptible and multidrug-resistant Mycobacterium tuberculosis clinical isolates and reference strains☆

Finding a gene or genes that are involved with multidrug resistance will be useful for finding a new target for the treatment of drug resistant tuberculosis. In this study, we aimed to compare the differences of the expression of 15 putative multidrug efflux pump genes in clinically isolated drug sensitive and multidrug resistant (MDR) Mycobacterium tuberculosis isolates, and reference strains. We found that these genes in the drug-sensitive and MDR M. tuberculosis isolates have similar rates of expressions. However, we found the expression levels of the all the genes are significantly higher in the clinical strains compared to the expression level of genes in the reference strains. In addition to this, it is found that standard strain has lower MIC value for the drugs including streptomycin and rifampin compared to the clinical isolate. We presume that the increase of the gene expression in the clinical strains is due to the exposure of antituberculosis drugs during treatment of patients, which cause constitutive expression of efflux systems, which might increase MIC levels of the major anti-tuberculosis drugs.

Epidemiology and Prevalence of Blastocystis spp. in North Cyprus

AbstractThis study was conducted to investigate the prevalence of Blastocystis spp. and its subtypes (STs) in North Cyprus; and to evaluate the presence of this parasite and its STs with respect to demographic, socioeconomic, and epidemiological factors, as well as gastrointestinal symptoms. Stool samples were collected from 230 volunteers. Each participant also filled out a questionnaire. The samples were examined microscopically by native-Lugol and trichrome methods and further tested by polymerase chain reaction (PCR) and sequencing. Prevalence of Blastocystis spp. infection was found to be 10.5%, 10.5%, and 27.8%, by direct microscopy, trichrome method, and PCR, respectively. No other parasites were detected in the specimens except Giardia spp. (n = 2; 0.8%) and Entamoeba coli (n = 1; 0.4%). The most common Blastocystis STs were ST3 (20; 31.2%), ST2 (18; 28.2%), ST1 (8; 12.5%), and ST4 (7; 11%); whereas other STs were identified as ST6 (3; 4.7%), ST7 (2; 3.2%), and non-ST (6; 9.4%). Presence of Blastocystis spp. and its STs was not significantly related to any of the demographic, socioeconomic, and epidemiological factors. Furthermore, no significant association of Blastocystis spp. and its STs with gastrointestinal symptoms was found. This study is the first investigation of the epidemiology of Blastocystis spp. in North Cyprus. Distribution of Blastocystis spp. and its STs among demographic, socioeconomic, and epidemiological factors showed complete homogeneity. Presence of the parasite and its STs was not significantly related with the gastrointestinal symptoms among symptomatic and asymptomatic individuals. These findings suggest that Blastocystis spp. may be part of the intestinal flora in humans.

Blastocystosis in post-traumatic splenectomized patients

BACKGROUND/AIMS The aim of this study was to determine the prevalence and significance of intestinal protozoa, specially Blastocystis spp., and to perform PCR-based subtype classification for understanding the importance of Blastocystis spp. in the pathogenesis of gastrointestinal disorders in post-traumatic splenectomized patients. MATERIALS AND METHODS A total of 60 stool samples were obtained from 30 post-traumatic splenectomized patients and 30 healthy controls. Wet mounts, trichrome and Kinyoun acid-fast stained slides were prepared from the stool specimens. PCR was used for detecting the presence of Giardia spp., Entamoeba spp., Dientamoeba fragilis, Cryptosporidium spp., Blastocystis spp. Genotyping was realized by using Blastocystis hominis STS primers. RESULTS In both study groups, any helminth eggs and other protozoa except Blastocystis spp. were not detected by microscopy and PCR, and also bacterial cultures were negative. Only stool microscopy was positive for Blastocystis spp. in 30% (9 of 30) of splenectomized patients and in 13% (4/30) of healthy controls. PCR for Blastocystis spp. was positive in 40% (12 of 30), B. hominis genotypes were 20% (6/30): STS1 in 10% (3/30) and STS3 in 10% (3/30) of splenectomized patients. In healthy controls Blastocystis spp. was 13% (4/30) by PCR and genotypes of B. hominis was not detected. The difference between the prevalence of Blastocystis spp. infection in splenectomized patients and control groups was statistically significant (p=0.020). Abdominal pain was the most frequent gastrointestinal symptom (p=0.019) among splenectomized patients positive for Blastocystis spp. CONCLUSION In post-traumatic healthy splenectomized patients, Blastocystis spp. were found to be the most prevalent protozoa and may be responsible for the gastrointestinal disorders.

Bacterial and protozoal pathogens found in ticks collected from humans in Corum province of Turkey

Background Tick-borne diseases are increasing all over the word, including Turkey. The aim of this study was to determine the bacterial and protozoan vector-borne pathogens in ticks infesting humans in the Corum province of Turkey. Methodology/Principal findings From March to November 2014 a total of 322 ticks were collected from patients who attended the local hospitals with tick bites. Ticks were screened by real time-PCR and PCR, and obtained amplicons were sequenced. The dedected tick was belonging to the genus Hyalomma, Haemaphysalis, Rhipicephalus, Dermacentor and Ixodes. A total of 17 microorganism species were identified in ticks. The most prevalent Rickettsia spp. were: R. aeschlimannii (19.5%), R. slovaca (4.5%), R. raoultii (2.2%), R. hoogstraalii (1.9%), R. sibirica subsp. mongolitimonae (1.2%), R. monacensis (0.31%), and Rickettsia spp. (1.2%). In addition, the following pathogens were identified: Borrelia afzelii (0.31%), Anaplasma spp. (0.31%), Ehrlichia spp. (0.93%), Babesia microti (0.93%), Babesia ovis (0.31%), Babesia occultans (3.4%), Theileria spp. (1.6%), Hepatozoon felis (0.31%), Hepatozoon canis (0.31%), and Hemolivia mauritanica (2.1%). All samples were negative for Francisella tularensis, Coxiella burnetii, Bartonella spp., Toxoplasma gondii and Leishmania spp. Conclusions/Significance Ticks in Corum carry a large variety of human and zoonotic pathogens that were detected not only in known vectors, but showed a wider vector diversity. There is an increase in the prevalence of ticks infected with the spotted fever group and lymphangitis-associated rickettsiosis, while Ehrlichia spp. and Anaplasma spp. were reported for the first time from this region. B. microti was detected for the first time in Hyalomma marginatum infesting humans. The detection of B. occultans, B. ovis, Hepatozoon spp., Theileria spp. and Hemolivia mauritanica indicate the importance of these ticks as vectors of pathogens of veterinary importance, therefore patients with a tick infestation should be followed for a variety of pathogens with medical importance.

Identification of methicillin-resistant Staphylococcus aureus carrying an exfoliative toxin A gene encoding phage isolated from a hospitalized patient in Turkey

From the four known isoforms of the staphylococcal exfoliative toxins (ETs), only ETA and ETB are the major causative agents. General knowledge is that the gene for ETA is located on the chromosome, whereas that for ETB is located on a large plasmid. Yoshizawa and co-workers (2000, Microbiol. Immunol. 44(3): 189-191) isolated, for the first time, a temperate phage (φETA) that carried the structural gene for ETA from an ETA-producing strain of Staphylococcus aureus. In this study, we presented eta gene encoding temperate phages isolated from methicillin-resistant S.aureus (MRSA) isolates obtained from patients in a Turkish hospital. Molecular analysis of the phage genome revealed that the eta gene is located upstream to amidase and holin genes, the same as in the φETA genome. However, partial sequence analysis of amidase and holin genes revealed polymorphic variation. In addition to polymorphic variation, restriction fragment length polymorphism (RFLP) analysis of all of the phage genomes showed that the ETA-containing phage is different from the rest of the phage genomes. The phylogenetic dendrogram of pulsed field gel electrophoresis (PFGE) analysis showed that the ETA-carrying MRSA is quite different from the rest of the MRSA strains. This is the first report showing that a MRSA strain carries an ETA-encoding phage.

The effect of splenectomy on the levels of PCV-13-induced memory B- and T cells

Splenectomised patients are associated with lifelong risk of fatal overwhelming post‐splenectomy infection (OPSI), which is mostly caused by Streptococcus pneumoniae. Today OPSI cases can still be reported even in patients with appropriate vaccination. In our study, the levels of vaccine‐specific memory B‐ and T cells were compared between control and splenectomised patients to enlighten the underlying reason.