Soykan Ozkoc

Identification of Acanthamoeba genotype T4 and Paravahlkampfia sp. from two clinical samples

In this study, two free-living amoebae strains, Acanthamoeba genotype T4 and Paravahlkampfia sp., which were isolated from keratitis cases are presented. While the Acanthamoeba strain was isolated as a single agent, the Paravahlkampfia strain was found together with herpes simplex virus. Neither of the patients were contact lens wearers, but they did have a history of minor corneal trauma. Amoebae were detected on non-nutrient agar covered with Escherichia coli. Based on PCR-amplified 18S rRNA-gene analysis the first isolate was identified as Acanthamoeba genotype T4 and the second as Paravahlkampfia sp. In thermotolerance tests, the maximum temperature at which trophozoites continued to divide was determined as 37 degrees C for this Acanthamoeba strain and 35 degrees C for the Paravahlkampfia strain. To the best of our knowledge, the Acanthamoeba strain described herein is the second molecularly identified Acanthamoeba strain in an Acanthamoeba keratitis patient in Turkey. However, the Paravahlkampfia isolate is believed to be the first strain that has been isolated from a keratitis patient and has been molecularly differentiated from Vahlkampfia.

The epidemiology research of Blastocystis hominis in the Dokuz Eylül University Medical Faculty Hospital between 2005 and 2009

OBJECTIVE An investigation of Blastocystis hominis (B. hominis) prevalance in 17756 patients with gastrointestinal system complaints who presented at the parasitology laboratory of the Dokuz Eylul University Medical Faculty Hospital between January 2005 and December 2009 was carried out. METHODS Fecal samples of all patients were examined using the native-Lugol and trichrome and Kinyoun acid-fast staining method after sedimentation in fecal concentration tubes. RESULTS One or more parasites were detected in 1510 (8.50%) of the patients. The distribution of the intestinal parasites was as follows: B. hominis 778 (4.38%), nonpathogenic amoebas 343 (1.93%), Giardia intestinalis (G. intestinalis) 205 (1,15%), Enterobius vermicularis (E. vermicularis) 46 (0.25%), Entamoeba histolytica/Entamoeba dispar (E. histolytica/E. dispar) 34 (0.19%), and other rare parasites 104 (0.58%). The most frequently seen parasite was B. hominis in fecal samples of patients with gastrointestinal complaints in our study. Distribution of 778 patients with B. hominis due to parasite forms was determined as: vacuolar in 525 (67.49%), granular in 115 (14.78%), both vacuolar and granular in 138 (17.73%) cases. CONCLUSION As B. hominis was the most frequently seen parasite in patients with gastrointestinal complaints, we suggest that the parasite should be considered as pathogenic and sufficient attention must be paid in routine stool examinations.

THE PREVALENCE OF HEAD LICE INFESTATION IN SCHOOL CHILDREN IN IZMIR, TURKEY

To the Editor: In regard to the paper by Ginter-Hanselmayer et al (1), an open clinical trial describing the therapeutic response to itraconazole in the treatment of tinea capitis, caused by Microsporum canis , there are some points that should be considered. The title of the paper, “Experience in a large cohort,” suggests that it was an observational study and that there was no intervention. However, it was conducted as an open trial. This trial did not have a control group, so it is not possible to conclude that itraconazole is effective in the treatment of tinea capitis. The Hawthorn effect is a potential bias in uncontrolled trials and could explain the results (2). Gupta et al (3) conducted a multicenter randomized, single-blinded, nonindustrysponsored trial and showed that terbinafine, itraconazole, and fluconazole had efficacy similar to that of griseofulvin in the treatment of tinea capitis. However, tinea caused by Microsporum sp. was not investigated in that study. In developing countries such as Brazil, where the prevalence of this disease is high, griseofulvin might be the choice for treatment because of its efficacy and cost, as it is much cheaper than itraconazol. Therefore, we need more convincing evidence of effectiveness, adverse effects, and even costs, in order to encourage us to change our first-line treatment for tinea capitis.

Prevalence of Pneumocystis jirovecii colonization in autopsy cases in Turkey.

Detection of Pneumocystis jirovecii and its DNA in clinically asymptomatic people is defined as colonization. The aim of this study was to reveal the colonization prevalence of P. jirovecii and affecting factors in an immunocompetent population. The study included 200 cases undergoing forensic autopsy between February 2015 and April 2015. The cause of death was non-medical conditions (group 1) in 111 cases (55.5 %), medical conditions (group 2) in 73 cases (36.5 %) and undetermined (group 3) in 16 cases (group 3). Tissue specimens about 1 g in weight were taken from the right upper pulmonary lobe. After DNA extraction, nested PCR targeting mitochondrial large subunit rRNA was used to detect P. jirovecii. Of 200 cases, 37 (18.5 %) had P. jirovecii DNA. There was not a significant difference in place of living, gender, smoking status and medication use between the cases with P. jirovecii and those without P. jirovecii. A significantly high rate of P. jirovecii colonization was detected in group 2 (χ²=7.674; P=0.022). P. jirovecii-colonized cases also had a chronic disease in 2 of 13 (group 1), 12 of 20 (group 2) and 1 of 4 (group 3) cases (χ²=5.571; P=0.062). A significantly high rate of the cases aged 0-1 year had P. jirovecii (5/11; 45.5 %) (χ²=5.639; P=0.018). The results of the study suggest that infants and patients with chronic diseases like cardiac or pulmonary diseases can be at risk for P. jirovecii colonization.

[Acute appendicitis and coinfection with enterobiasis and taeniasis: a case report].

Parasites are rarely associated with inflammation of the appendix. Generally, parasites cause acute abdominal pain via blocking the gut lumen. In this article, we presented a case of appendicitis where Enterobius vermicularis was detected in the surgical specimen and Taenia was detected in the stool. A 31 year old male patient was admitted to the emergency room with severe abdominal pain, which has begun two days ago. On physical examination, tenderness was positive on palpation of the right lower abdominal quadrant and the patient was operated on with the diagnosis of acute appendicitis. Histopathological examination of the patients appendectomy material revealed numerous parts of parasites resembling Enterobius vermicularis and slight mucosal erosion. On parasitological examination of the patients stool, Taenia eggs and adult forms were determined. Antiparasitic therapy was started with niclosamide for taeniasis and albendazole for enterobiasis. Parasitic infections can mimic acute appendicitis clinically. Radiological and laboratory findings do not help to distinguish the diagnosis of acute appendicitis. In the histopathological examination of the appendix, the findings of acute inflammation of the appendix wall may not be defined. For patients with normal histopathological examination, screening for parasites should be done, and anti-parasitic treatment should be started after appendectomy.

Evaluation of pulmonary microsporidiosis in iatrogenically immunosuppressed patients.

INTRODUCTION Microsporidia spp. are ubiquitous and infect a wide variety of intervertebrates and vertebrates, including humans. Pulmonary microsporidiosis, characterized by nonspecific symptoms like fever, cough and dyspnea, is often overlooked in the differential diagnosis of pulmonary infections in immunsupressed patients. In this study, we aimed to determine the prevalence of pulmonary microsporidiosis in iatrogenically immunosuppressed patients and to evaluate the patient characteristics. MATERIALS AND METHODS Bronchoalveolar lavage (BAL) specimens from 63 iatrogenically immunosuppressed patients and 28 controls were examined with PCR. By using PMP1 and PMP2 common primers specifically designed for Enterocytozoon bieneusi, Encephalitozoon intestinalis, Encephalitozoon cuniculi and Encephalitozoon hellem small-subunit ribosomal DNA (SSU-rDNA) regions at 250-279 bp were amplified. In addition, PCR positive BAL specimens were examined with modified trichrome staining method for Microsporidia spores. RESULT Out of 63 immunosuppressed patients, nine (14.2%) had Microsporidia spp., but none of the control patients had Microsporidia spp. on PCR. This difference between two groups was statistically significant (χ² =4.439; p=0.035). On the other hand there was not a statistically significant relationship between PCR positivity and patient characteristics such as gender and age. Of nine patients with Microsporidia PCR positive, only one had spores of Microsporidia sp. Out of eight patients without spores, one had Mycobacterium tuberculosis, one patient had Klebsiella pneumoniae and five patients had Pneumocystis jirovecii DNA. CONCLUSION This is the first study to evaluate the pulmonary microsporidiosis in immunosupressed patients in Turkey. The results of the study indicated that Microsporidia spp. should be taken into account in the differential diagnosis of pulmonary infections in immunosupressed patients and it is important to use molecular methods such as PCR in the laboratory diagnosis of the causative agent.

Investigation of Pneumocystis jirovecii colonization in patients with pulmonary diseases

OBJECTIVE The detection of Pneumocystis jirovecii or its DNA in respiratory samples from individuals who do not have signs or symptoms of pneumonia has been defined as colonization. In this study, we aimed to investigate the prevalence of P. jirovecii colonization in patients with various lung diseases. METHODS Thirty patients who were followed-up and who had undergone bronchoscopy for diagnosis of different underlying diseases or pulmonary signs were included in the study. Bronchoalveolar lavage (BAL) fluids of these patients were analyzed with nPCR amplification of the mt-LSUrRNA gene of P. jirovecii. In addition to nPCR, giemsa, Gomoris methenamine silver (GMS), and indirect fluorescence antibody (IFA) staining assays were applied to all samples. RESULTS P. jirovecii DNA was detected in 21 of 30 (70%) BAL samples by nPCR. However, P. jirovecii cysts were found in 1 of 21 nPCR-positive samples by giemsa and GMS. IFA assay showed six samples to be positive, but only four of them were found to be positive by nPCR. CONCLUSION Result of our study showed that prevalence of P. jirovecii colonization is particularly high in patients with chronic pulmonary diseases, and nPCR was a good assay for evaluation of the colonization of P. jirovecii.

Pneumocystis pneumonia in a renal transplant recipient.

Pneumocystis pneumonia (PCP) is an opportunistic infection caused by Pneumocystis jirovecii (P. jirovecii) in humans. We reported a 23 year-old male patient who developed pneumonia after renal transplantation. P. jirovecii cysts and trophozoites were detected in bronchoalveolar lavage (BAL) samples of the patient by Giemsa, methenamine-silver and Toluidine-O staining. The patient, who was diagnosed as PCP, was discharged as he recovered by 21 days trimethoprim-sulfamethoxazole (TMP-SMX) therapy. This case, who developed PCP even though he had received prophylaxis after transplantation, was reported to emphasize the importance of the agent in immunocompromised patients.

Pneumocystis jirovecii Pneumonia in a Patient with Interstitial Lung Disease

Pneumocystis pneumonia (PCP) caused by Pneumocystis jirovecii (P. jirovecii) is an opportunistic pulmonary infection that occurs in immunocompromised patients. Here, a 49-year-old female patient who was admitted to our hospital with respiratuary distress and whose bronchoalveolar lavage (BAL) fluid specimens had P. jirovecii and Aspegillus fumigatus was presented. She had been treated with corticosteroids because of interstisial lung disease and she was also diabetic. It is important to define the coinfection developed in the presence of immunosuppression.

Absence of dihydropteroate synthase gene mutations in Pneumocystis jirovecii strains isolated from Aegean region of Turkey

Sulfonamide group drugs and their antimetabolite combinations are the most preferred drugs in the treatment and prophylaxis of Pneumocystis jirovecii pneumonia (PCP). Especially with the long-term use of trimethoprim-sulfamethoxazole (TMP-SMX) and dapsone, certain point mutations in the Pneumocystis jirovecii (P. jirovecii) dihydropteroate synthase (DHPS) gene are known to play an important role in the development of resistance. In the present study, we investigated the 165th and 171st nucleotide mutations in the DHPS gene in the P. jirovecii isolates from immunosuppressed and immunocompetent cases. P. jirovecii isolates from the bronchoalveolar lavage fluid (BALF) samples of 31 hospitalized cases and lung tissue samples of 37 autopsy cases were included in the study. For the analysis of wild-type and mutant genotypes, after the touchdown-PCR amplification method, the restriction fragment length polymorphism (RFLP) method was used. In this study, P. jirovecii DHPS gene was amplified in 28 of 68 (41%) of the samples. The RFLP method revealed that all the isolates in which the DHPS gene was amplified were considered as wild-type genotypes. To our knowledge, this present study is the first study in Turkey investigating P. jirovecii DHPS gene mutations associated with the sulfonamide resistance. All the isolates showed a wild-type pattern indicating that the occurrence of P. jirovecii DHPS mutations in Turkey is very low or absent.