Investigation of Blastocystis hominis Frequency in Patients with Diabetes by Microscopy and Conventional PCR Methods.

Background: We aimed to determine the frequency and subtype of B. hominis in diabetic patients. Methods: One hundred and fifty diabetic patients and 100 healthy people without any chronic disease were included in the study. Stool samples were analyzed by native-Lugol, condensation, trichrome staining and PCR methods. Results: In 150 patients with diabetes; B. hominis was detected in 38 (25.3%) by PCR, in 34 (22.7%) by native-Lugol and trichrome staining. In the control group, 14 (14%) out of 100 subjects were positive by PCR, and 10 (10%) were positive by native-Lugol and trichrome staining. In the statistical evaluation, a significant difference was found between gender (P=0.023), age (P=0.045; ≤35 and >35 comparison), duration of diabetes (P=0.04), the HbA1c value (P=0.023; <8 and ≥8 comparison), and B. hominis positivity. ST1 was determined in 76.9% of patients with diabetes, and ST2 was determined in 23.07%. Considering the 3 methods, B. hominis positivity was detected in 40 patients (26.7%) in diabetic group and in 14 participants (14%) in the control group (P=0.011). Conclusion: B. hominis is a factor to be considered in patients with diabetes. Herein, the most common subtype detected in the patients with diabetes mellitus was ST1, but this result was not considered sufficient to reveal the importance of the subtype factor in the pathogenicity of B. hominis in patients with diabetes. In this context, there is a need for more comprehensive studies in both diabetic and other immunocompromised patient groups.

Investigation of the presence of Enterocytozoon bieneusi and Encephalitozoon intestinalis in immunosuppressed patients with diarrhea by IFA and real time PCR methods.

PURPOSE: This study aimed to investigate the frequency of Enterocytozoon bieneusi and Encephalitozoon intestinalis in patients with diarrhea in the immunosuppressed. METHODS: Patients between the ages of 18-85 who applied to different clinics of Mus Bulanik and Bitlis State Hospitals and were referred to the microbiology or parasitology laboratory were selected for this study. A total of 200 individuals, including 88 immunosuppressed with diarrhea patients, 38 immunocompetent with diarrhea patients, 38 immunosuppressed without diarrhea patients, and 36 immunocompetent without diarrhea individuals, were included. Collected stool samples were evaluated using IFA-MAbs and real-time PCR methods to determine the frequency of E.intestinalis and E.bieneusi. RESULTS: E. intestinalis was detected in 59 (29.5%) of 200 samples and E. bieneusi was detected in 46 (23.0%) of them. Mixed infection was detected in 16 (8%) of the positive samples. While there was no statistically significant difference between E. intestinalis positivity and gender, age, diarrhea status and immune system status, a statistically significant relationship was determined between E. bieneusi positivity and diarrhea. When the real-time PCR method was accepted as the gold standard, the sensitivity of the IFA-MAbs method in the diagnosis of E. intestinalis was 94.54%, the specificity was 97.24, the sensitivity in the diagnosis of E. bieneusi was 95.45%, and the specificity was 98.72%. The overall accuracy of the IFA-MAbs method was 96.5% for the diagnosis of E. intestinalis and 98% for the diagnosis of E. bieneusi. CONCLUSIONS: The findings suggest that E. intestinalis and E. bieneusi should be considered in both immunosuppressed and healthy individuals with diarrhea. IFA-MAbs method can be used in addition to the real-time PCR method to diagnose E. intestinalis and E. bieneusi.

Investigation of Pneumocystis jirovecii in Lung Cancer Patients with the Nested PCR Method

Objective: Pneumocystis jirovecii (P. jirovecii) is an opportunistic pathogen in humans. Early diagnosis and optimal treatment of patients with P. jirovecii pneumonia (PJP) remains a key priority. This study investigated P. jirovecii in patients with lung cancer using the nested-polymerase chain reaction (PCR) method and examined the relationship between P. jirovecii and clinical findings. Methods: The study included 60 patients with lung cancer and 30 patients without lung cancer. The bronchoalveolar lavage (BAL) fluid samples of these 90 individuals were taken for diagnostic purposes in the University of Health Sciences Turkey, Van Training and Research Hospital, Clinic of Chest Diseases. Patient information was recorded. After DNA isolation from the BAL fluid samples taken from patients, the nested-PCR protocol for amplification of mtLSUrRNA in P. jirovecii was performed. Results: P. jirovecii DNA was detected in 40 (66.67%) of the lung cancer patients included in the study and in six (20%) patients without lung cancer, that is, in 46 (51.11%) patients. The rate of nested-PCR positivity in the lung cancer group was significantly higher than that in the non-lung cancer group (p=0.0001). Additionally, a statistically significant correlation was found between anorexia and weight loss, fever and sputum P. jirovecii positivity in patients with lung cancer (p<0.005). Conclusion: These findings suggest that lung cancer patients should be evaluated for PJP.

Investigation of Intestinal and Blood Parasites in People Returning to Turkey with a History of Traveling Abroad During the Pandemic

Objective: To investigate intestinal and blood parasites in people who have a history of traveling abroad during the Coronavirus disease-2019 pandemic and returning to Turkey. Methods: In this study, 104 patients with gastrointestinal system and/or fever complaints who had traveled abroad during the pandemic period and returned to Turkey were included. Parasitic agents were investigated by taking blood and stool samples from the patients. Additionally, urine samples were obtained from patients with hematuria or dysuria with the suspicion of schistosomiasis. A direct microscopic examination, the Crypto-Giardia immunochromatographic test, and ELISA methods were used in the examination of the stool samples. In order to detect Plasmodium species, blood samples were examined by preparing both the rapid diagnostic test and thick drop and thin smear preparations. Results: One or more parasite species were detected in 38 (38.5%) of 104 patients included in the study. While intestinal parasites were detected in 16 (32%) of 50 patients who traveled to Iran and 16 (33.3%) of 48 patients who traveled to Northern Iraq, blood parasites were not found. Schistosoma mansoni was detected in all 5 of the patients with a history of traveling to Sudan. Plasmodium falciparum was detected in 1 patient who traveled to the African continent. Conclusion: It is vital to take precautions to prevent parasitic diseases, such as malaria and schistosomiasis, during travels to African countries. During travels to neighboring countries of Turkey, such as Northern Iraq and Iran, hygiene should be paid attention to, so as to prevent contracting intestinal parasitic diseases. In addition, it was concluded that people who plan to travel abroad should have information about the endemic parasitic diseases of the country that they are going to.

Genetic Diversity of Echinococcus granulosus Isolated from Humans: A Comparative Study in Two Cystic Echinococcosis Endemic Areas, Turkey and Iran.

Cystic echinococcosis (CE) is one of the most important zoonotic parasitic diseases caused by the larval stage of Echinococcus granulosus. Based on molecular studies and DNA sequencing, E. granulosus has been classified into 10 different genotypes (G1 to G10). Two neighboring countries, Turkey and Iran, are considered the two main foci of CE in the Middle East. The current study is aimed at examining the genotype diversity of E. granulosus isolated from human clinical samples in Turkey and Iran. Surgically removed human hydatid cysts were collected from East Azerbaijan and Fars provinces in Iran and Van province in Turkey. After extracting DNA, performing PCR, targeting the cox1 gene, the PCR products were purified from the gel and were sequenced from both directions. The sequences were aligned and compared, using BioEdit and also the BLAST program of GenBank. The maximum likelihood tree was constructed based on the Tamura-Nei model, using the MEGAX software. Phylogenetic analysis showed that the human isolated samples were classified into two major clades: G1 (from Iran and Turkey) and G3 (5 samples from northwestern Iran and one sample from Turkey). The mean and degree of genetic divergence (K2P) between the two major clades, G1 and G3, were 0.2% and 0.7 ± 0.4%, respectively. The findings of the current study revealed that the sheep strain (G1) and the less important strain G3 have major roles in the transmission cycle of CE in two neighboring countries, Iran and Turkey. Therefore, it is necessary to interpose the life cycle of this parasite and reduce the disease burden in livestock and humans by adopting common regional preventive and control policies.

Molecular characterization of human lung and liver cystic echinococcosis isolates in Van Province, Turkey.

Cystic echinococcosis (CE) is a zoonotic infection and could lead to significant public health problems. The genetic diversity of CE includes five species: E. granulosus sensu stricto (s.s.) (G1-G3), Echinococcus equinus (G4), Echinococcus ortleppi (G5), Echinococcus canadensis genotypic cluster (G6, G7, G8 and G10, with the doubtful G9) and the Echinococcus felidis (lion strain). The species are important in epidemiology, pathology, control, prevention measures and vaccine/drug designs. The aim of the present study was to determine the E. granulosus genotypes in humans in the Van province in east of Turkey. In total, 102 echinococcal cysts were collected from operated patients. Genomic analyses were conducted with PCR-RFLP of the rDNA internal transcribed spacer 1 (ITS1) fragment and partial PCR sequencing of the cytochrome c oxidase subunit 1 (cox1) mitochondrial DNA gene region. In total, DNAs of 96 isolates could be extracted, unfortunately six extractions failed. The PCR-RFLP analysis findings were identical in all isolates. Two bands were observed at approximately 300 bp and 600 bp. All profiles corresponded to the G1-G3 strain. Also, 446 bp amplified gene regions were observed for cox1. Out of 20 samples, alignment of 16 sequences exhibited a total identification (100%) of granulosus sensu stricto (G1/G3). Of 16 samples, 8 were obtained in the lung and 12 were obtained in the liver; 8 belonged to male and 12 belonged to female patients. Other four samples exhibited one nucleotide substitution at different positions. Four samples had one nucleotide substitution at different positions. We detected single nucleotide variations in TRH1, TRH67, TRH85 and TRH89 isolates at the positions C240T; G330T; G211A and T157C, respectively. In conclusion, the present study was the first comprehensive molecular investigation on genetic characterization of human CE isolates in Van region. The findings demonstrated that E. granulosus s.s. was the dominant species, which indicated that the sheep-dog cycle was the source in human infections. And, probably, it would be possible to describe these mutations as "Turkey" or "lung" variants. In addition to contributing molecular epidemiological data, the present results should be considered when designing and implementing E. granulosus control programs.

A Rare Case of Secondary Hydatid Cyst: Uterus and Colon Locations in the Same Patient

The aim of the present study was to present a case with secondary hydatid cysts in both uterus and colon. The patient was a 71-year-old female living in Hakkari, Turkey. She was admitted to the Van Yuzuncu Yil University Faculty of Medicine Medical Center with complaints of chronic abdominal and pelvic pain, and swelling in the abdomen. First, the sagittal T2 weighted magnetic rezonance imaging (MR) showed a type-3 cyst hydatid with daughter vesicles located at the posterior of uterus. Later, MR revealed a type-2 cystic lesion with detached membrane adhered to the anterior wall of colon and it was reported to be associated with abdomen. When the previous liver surgery history of the patient was kept in mind, the new finding was suggestive of a secondary cystic hydatid . In conclusion, it is possible to diagnose secondary cystic echinococcosis in patients with a history of primary cyst surgery in liver or any other organ by combining the symptoms and imaging findings.