Genetic diversity of Pneumocystis jirovecii isolates among Turkish population based on mitochondrial large subunit ribosomal RNA and dihydropteroate synthase gene typing.

Pneumocystis jirovecii (P. jirovecii) is an atypical fungus that can cause severe interstitial pneumonia in immunocompromised patients. In this study, mitochondrial large subunit ribosomal RNA (mtLSU-rRNA) and dihydropteroate synthase (DHPS) gene polymorphism in P. jirovecii isolates were investigated in Western Turkey's Izmir province and its surroundings. For this purpose, a total of 157 P. jirovecii isolates obtained from bronchoalveolar lavage samples of hospitalized cases and lung tissue samples of autopsy cases who died outside hospital were examined. Genotypes were identified by direct sequencing of mtLSU-rRNA restriction fragment length polymorphism analysis of the DHPS gene amplicons. The mtLSU-rRNA analysis revealed that genotype 2 was the most common genotype with 58%. The following genotypes were genotype 3 (13%), genotype 1 (11.6%) and genotype 4 (5.1%), while genotype 5 (0.7%) was detected in only one autopsy case. In addition, 16 (11.6%) cases had dual or triple different genotypes (mixed infection). It was observed that the genotype distribution was not affected by characteristics such as age, gender and immune status. However, the predominance of genotype 2 in solid organ tumors and the predominance of mixed infection in patients with chronic pulmonary disease were statistically significant. On the other hand, DHPS gene amplification was positive in 137 (87.3%) of 157 samples. While no mutation was observed in 135 samples, the association of wild-type and 57th codon mutation was detected in two hospitalized cases (1.5%). In this study, important epidemiological data on the distribution of mtLSU-rRNA genotypes were obtained. Also the existence of DHPS gene mutations associated with potential drug resistance in our community was shown for the first time. Further studies are needed to evaluate the possible effects of genotypes on the prognosis of the disease to help with the clinician's treatment decisions. LAY ABSTRACT: Pneumocystis jirovecii (P. jirovecii) is an atypical fungus that can cause life-threatening pneumonia in immunocompromised patients. In this study, we investigated the mtLSU-rRNA and DHPS gene polymorphisms in P. jirovecii isolates from both hospital and autopsy cases.

Pneumocystis jirovecii Pneumonia in Solid-Organ Transplant Recipients: A National Center Experience.

OBJECTIVES: Immunosuppressive therapies have impro-ved survival in solid-organ transplant recipients at the expense of increased prevalence of opportunistic infections. We investigated the prevalence, risk factors, and prognosis of Pneumocystis jirovecii pneumonia in solid-organ transplant recipients who were followed by our transplant unit. MATERIALS AND METHODS: We conducted a retrospective observational study using medical record reviews to identify all adult solid-organ transplant recipients who underwent bronchoscopy and bronchoalveolar lavage between January 2011 and 2018. We collected clinical characteristics, including risk factors and prognosis. Pneumocystis jirovecii pneumonia symptoms com-patible with microscopy and/or positive nucleic acid amplification assays were defined as proven infection by P. jirovecii pneumonia. RESULTS: We identified 312 adult solid-organ transplants (114 renal, 1 kidney and pancreas, 197 liver) in this period. Overall, 113 (36.2%) pulmonary disease consultations were performed in the posttransplant stage, and 46 (40.7%) patients underwent bronchoalveolar lavage with P. jirovecii screening. We identified 18 patients who tested positive for P. jirovecii infection according to nucleic acid amplification assay; 3 were not proven, and 7 had a transplant date before 2011. The prevalence was 8/312 (2.6%); of these 8 patients, 5 had the same genotype cluster. Median P. jirovecii pneumonia development time was longer in renal transplant recipients (P = .016). Only renal transplant recipients were offered Pneumocystis prophylaxis for 6 months. Concomitant viral infection including cytomegalovirus was the only significant factor for P. jirovecii pneumonia development (P = .028). Intensive care admission was 40% (n = 6), and disease-related mortality was 33% (n = 5). CONCLUSIONS: The overall prevalence of P. jirovecii pneumonia in solid-organ transplant recipients was similar to other single-center reports. Prophylaxis prevented early posttransplant P. jirovecii pneumonia. However, P. jirovecii pneumonia may develop at any posttransplant stage, and viral infections other than cytomegalovirus should also be considered as a predictor.

[Evaluation of Real-Time Polymerase Chain Reaction in Pneumocystis jirovecii Laboratory Diagnosis]. / Pneumocystis jirovecii Laboratuvar Tanisinda Gerçek Zamanli Polimeraz Zincir Reaksiyonu Yönteminin Degerlendirilmesi.

Pneumocystis jirovecii is a human-specific species and causes fatal infections like P.jirovecii pneumonia (PCP) in immunocompromised persons. Although direct microscopy is the gold standard in the diagnosis of the microorganism, molecular methods such as polymerase chain reaction (PCR) are needed in non-human immune deficiency virus (HIV) immunosuppresive patients with low P.jirovecii burden. In this study, we aimed to evaluate the value of real-time PCR (Rt-PCR) in the laboratory diagnosis of P.jirovecii. Bronchoalveolar lavage (BAL) specimens of 658 patients sent to Dokuz Eylul University Hospital Central Medical Parasitology Laboratory on suspicion of PCP were included in the study. BAL fluids were evaluated for identification of P.jirovecii mitochondrial gene coding ribosomal large subunit (mtLSUrRNA) using Rt-PCR. In addition, Giemsa and Gomori's methenamine silver (GMG) staining assays were applied to all samples and nested PCR (n-PCR) assay was applied to positive samples detected by real time PCR. Ninety-two (14.3%) of these samples were positive by Rt-PCR. Of these 92 patients, 85 (92.4%) were positive with n-PCR. Only seven of the specimens had P.jirovecii cysts and trophozoites with microscopic examination. The mean cycle threshold (CT ) value of Rt-PCR positive patients was 29.7 (18.17 ≤ CT ≤ 37.96). P.jirovecii load in these patients was calculated as 2.6 x 101-6.15 x 107 copies/ml. The difference between the mean CT values of n-PCR positive and negative results was statistically significant (p< 0.01). The CT values of Rt-PCR of the samples with positive microscopy were; 18.2, 20.9, 22.2, 24.3, 24.7, 26.5, 29.7. The difference between the CT means of the samples with positive and negative microscopy was statistically significant (p< 0.05). When positive patients were grouped according to their diagnosis; the lowest mean CT value (CTmean= 24.8) was found in HIV-positive patients. On the other hand, CT values were found to be significantly lower in the organ transplantation patients (CTmean= 26.15) and in the collagen-vascular-inflammatory patient group (CTmean= 27.8). This study demonstrated that Rt-PCR was the effective method in the diagnosis of P.jirovecii in the laboratory. Conventional n-PCR method was found to be more unsuccessful than Rt-PCR in the presence of very low density organism; direct microscopy is generally found to be positive in samples with a higher burden of P.jirovecii.

Distribution of Intestinal Parasites Detected between September 2011-2018 at Dokuz Eylül University Medical Faculty Hospital.

OBJECTIVE: Intestinal parasitic diseases are important public health problems in our country as well as in the world. In this study, intestinal parasites were investigated in patients admitted to Dokuz Eylül University Hospital (DEUH) with various gastrointestinal system complaints. METHODS: Patients (n=18460) who were referred to the DEUH Central Parasitology Laboratory between January 2011 and December 2018, were included in the study. Fecal samples were examined with Nativ-lugol method and then formol ethyl-acetate precipitation method was applied. Trichrome and kinyoun acid-fast stainings were performed on the necessary samples. Demographic data of the patients were obtained from the hospital's and laboratory's information operating system. RESULTS: One or more parasites were detected in 6% (1128) of 18460 patients examined. The mean age of the patients with parasites was 39.7 (±23.1) years, of which 53.3% were male and 47.6% were female. The distribution of parasites detected were as follows; 4.8% (879) Blastocystis hominis, 0.7% (135) amoebas other than Entamoeba histolytica/dispar, 0.4% (70) Giardia intestinalis, 0.3% (49) Enterebius vermicularis, 0.1% (21) Entamoeba histolytica/dispar, and 0.01% (10) other rare parasites. CONCLUSION: Our study shows that intestinal parasitic infections are still an important public health problem in our region and that there is a decrease in their incidence.

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.

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.

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 parasitic and viral pathogens in mussels (Mytilus galloprovincialis) in the Gulf of Izmir, Turkey.

OBJECTIVES: To investigate Microsporidia spp. parasite, hepatitis A virus (HAV), and norovirus (NoV) contamination in mussels collected from 8 stations in the inner, middle, and outer regions of the Gulf of Izmir. METHODS: In this cross-sectional study carried out between August 2009 and September 2010 in the Gulf of Izmir, Turkey, 15 mussels collected from each of the stations each season were pooled and homogenized to create a single representative sample. Thirty representative samples were available for analysis. Direct polymerase chain reaction (PCR), RT-nested PCR, and RT-booster PCR were used to investigate the pathogens.  RESULTS: The mussels were negative for Microsporidia spp., but 8 (26.7%) samples analyzed were positive for HAV and 9 (30%) were positive for NoV. Excluding Foca and Gediz, viral contamination was detected in all of the stations sampled.  CONCLUSION: Our results suggest that viral contamination is present in mussels in the Gulf of Izmir and may pose a potential threat to human health in the region. Necessary measures should be taken to prevent future illness due to these pathogens.

[Investigation of the in vitro effects of Melissa officinalis L., Mentha x piperita L. and Ocimum basilicum L. (Lamiaceae) essential oils on the cysts and trophozoites of Acanthamoeba castellani]. / Melissa officinalis L., Mentha x piperita L. ve Ocimum basilicum L. (Lamiaceae) uçucu yaglarinin Acanthamoeba castellani kist ve trofozoitleri üzerine in vitro etkisinin arastirilmasi.

Acanthamoeba species are free living amoeba found widely all over the world. They are responsible for Acanthamoeba keratitis (AK), an infection which is especially seen in contact lens users and after minor corneal traumas, that may lead blindness. At present, antifungals and antiseptics are used for the treatment of AK cases, however, some problems such as long treatment periods and the occurrence of side effects, resistance of cyst forms against drugs, emphasize the need for new drugs. There are some published studies that pointed out the effectiveness of plant extracts and essential oils on Acanthamoeba spp. The aim of this study was to investigate the in vitro effects of essential oils of Mentha x piperita L. (peppermint), Melissa officinalis L. (lemon balm) and Ocimum basilicum L. (basil) belonging to Lamiaceae family, on the cysts and trophozoites of Acanthamoeba castellanii. The strain used in our study, namely A. castellanii T4 genotype, is the most frequently isolated amoeba from environment and also the causative agent of AK and granulomatous amebic encephalitis. For the determination of amebicidal activity, essential oils obtained from Mentha x priperita L., Melissa officinalis L. and Ocimum basilicum L. by Neo-Clevenger type of distillation apparatus have been used. In vitro experiments were performed by using 96-well microplates. Cyst and trophozoite solutions were added on the essential oil dilutions to obtain the last concentrations of 40, 20, 10, 5, 2.5 and 1.25 µg/ml for the cysts, and 10, 5, 2.5, 1.25, 0.625 and 0.313 µg/ml for the trophozoites. After the incubation of microplates at 30oC for 1, 6, 24, 48 and 72 hours, the viability of parasitic forms were evaluated under the light microscope followed by staining trypan blue. It was found that, each essential oil showed amebicidal effect on A.castellani cysts and trophozoites dependent on dosage and time, when compared with the control group, The maximum lethal effect occured with Melissa officinalis followed by Mentha x piperita and Ocimum basilicum, respectively. In our study, susceptibility of A.castellanii trophozoites to essential oils were more than the cysts, as expected. The essential oils of Melissa officinalis and Mentha x piperita showed 100% lethal effect at their highest concentrations whereas the essential oil of Ocimum basilicum showed only 63.3% lethal effect on cysts after 72 hours at the highest concentration (40 µg/mL). The results of this first study investigating the activities of essential oils extracted from Mentha x piperita, Melissa officinalis and Ocimum basilicum against Acanthamoeba spp. cysts and trophozoites, have suggested that, these essential oils could be potential novel and alternative natural products for the treatment of Acanthamoeba spp. infections.

[Investigation of Pneumocystis jirovecii pneumonia and colonization in iatrogenically immunosuppressed and immunocompetent patients]. / Iyatrojenik immünosüpresif ve immünokompetan hastalarda Pneumocystis jirovecii pnömonisi ve kolonizasyonunun arastirilmasi.

Pneumocystis pneumonia (PCP) is a potentially life-threatening infection for the immunocompromized patients. However, Pneumocystis jirovecii colonization can also be detected in healthy individuals and in patients with various underlying lung diseases. The aim of this study was to evaluate the immunocompetent and iatrogenically immunosuppressed patients in terms of PCP and P.jirovecii colonization. A total of 92 patients (66 male, 26 female; age range: 18-93 years, median: 58.5) who underwent bronchoscopy due to various pulmonary symptoms between January 2011-April 2014, were included in the study. Of these patients, 65 were under immunosuppressive therapy (38 were treated with anti-cancer drugs, 15 with anti-rejection/immunomodulatory drugs and 12 with corticosteroids), while 27 were immunocompetent. Bronchoalveolar lavage (BAL) fluids were evaluated for the presence of P.jirovecii mitochondrial gene coding ribosomal large subunit (mtLSUrRNA) with nested PCR (nPCR) method. All of the samples were also examined by Giemsa and Gomori's methenamine silver (GMG) staining methods. P.jirovecii DNA was detected in 31 (33.7%) out of 92 BAL samples by nPCR. Although six immunosuppressed patients were positive in the first round of amplification, 26 of 65 (40%) immunosuppressed and five of 27 (18.5%) immunocompetent patients were positive with nPCR. P.jirovecii cysts and trophozoites were detected in only five (16.1%) of the 31 nPCR positive samples. The probability of being immunosuppressive among nPCR positive cases was statistically higher than nPCR negative cases (χ²= 3.940; p= 0.047). This difference was more significant in organ transplant recipients and patients under anti-rejection/immunomodulatory treatment (χ²= 6.715, p= 0.01; χ²= 5.550, p= 0.018, respectively). When clinical, laboratory and radiological findings of nPCR positive patients were considered, five patients (2 kidney transplant, 1 bone marrow transplant, 1 interstitial lung disease and 1 lung cancer case) in immunosuppressed group were interpreted as "definite PCP" and eight patients (2 kidney transplant, 1 leukemia, 1 connective tissue disease, 1 Wegener's granulomatosus, 2 rheumatoid arthritis and 1 lung cancer case) were interpreted as "probable PCP". Other 18 (19.6%) nPCR positive patients, of them 13 were immunosuppressive and five were immunocompetent, were considered as "P.jirovecii colonization". The colonization rate was determined as 50% (13/26) in immunosuppressive patients, and was mostly detected in patients with hematological malignancies (4/13), followed by patients with solid tumors (3/13) and organ transplantations (3/13). On the other hand, all of the nPCR positive immunocompetent patients (5/5) were evaluated as colonization. In this study significant data was obtained about P.jirovecii epidemiology in our country. Our results also showed that iatrogenically immunosuppressed patients are under risk of PCP and nPCR method is more sensitive than conventional PCR and classical staining methods in the diagnosis of these patients.

[Pneumocystis jirovecii Pneumonia in a Patient with Interstitial Lung Disease]. / Interstisyel Akciger Hastaligi Zemininde Gelisen Pneumocystis jirovecii Pnömonisi.

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.

[Acute appendicitis and coinfection with enterobiasis and taeniasis: a case report]. / Akut apandisit ve enterobiyaz ile taeniyaz: bir olgu sunumu.

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 patient's appendectomy material revealed numerous parts of parasites resembling Enterobius vermicularis and slight mucosal erosion. On parasitological examination of the patient's 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.

[Investigation of Pneumocystis jirovecii colonization in patients with pulmonary diseases]. / Akciger Hastaligi Olan Hastalarda Pneumocystis Jirovecii Kolonizasyonunun Arastirilmasi.

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, Gomori's 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.

[Evaluation of Trichinella cross-reactions in the serological diagnosis of toxocariasis]. / Toksokariyazis Serolojik Tanisinda Trichinella Çapraz Reaksiyonlarinin Degerlendirilmesi.

Toxocariasis caused by the nematode larvae of the Toxocara genus is a worldwide parasitic zoonosis. Diagnosis of human toxocariasis commonly relies on serological tests since the symptoms and signs of Toxocara infection are not pathognomonic. However Toxocara larval excretory-secretory (TES) antigen used in serological tests may exhibit low specificity due to the cross-reactions between related helminth infections such as ascariasis, anisakiasis, strongyloidosis and filariasis. In this study, we aimed to evaluate the possible effect of Trichinella cross-reactions in the serological diagnosis of toxocariasis by using ELISA and Western blot (WB) assay. For this purpose, sera samples of 209 trichinellosis patients who were definitely diagnosed during the Trichinella britovi outbreak occurred in Izmir in January 2004, were used. All the samples were screened initially by commercial Toxocara IgG-ELISA kit (Cypress Diagnostics, Belgium), then commercial Toxocara IgG-WB (Test-Line Diagnostics, Czech Republic) was applied to positive/ borderline-positive sera for confirmation. In our study, 94.3% (197/209) of the sera were found seronegative, while nine were positive and three were borderline. Thus a total of 12 (5.7%) sera were considered as seropositive by Toxocara IgG-ELISA. According to the results of WB, only one sera with the antigenic bands of 120 kDa, 32 kDa and 26 kDa in molecular weights was evaluated as positive. Four sera samples were found to be borderline. In three of border sera, the antigenic bands of 120 and 70 kDa in molecular weights were observed together and one sera had three (120, 70 and 32 kDa) different antigenic bands. Seven sera that had been found to be positive by ELISA was considered as negative by WB. While no bands was observed in four of these, three samples had an antigenic band of 120 kDa which had no diagnostic value when it was found alone. The results of our study showed that the crossreactivities between anti-Trichinella antibodies and TES antigens may be observed during Toxocara IgG ELISA assay. For that reason the positive Toxocara IgG-ELISA result should be confirmed by different tests such as WB for the definitive diagnosis of toxocariasis.

[The epidemiology research of Blastocystis hominis in the Dokuz Eylül University Medical Faculty Hospital between 2005 and 2009]. / Dokuz Eylül Üniversitesi Hastanesi'ne 2005-2009 Yillari Arasinda Basvuran Olgularda Blastocystis hominis Epidemiyolojisinin Arastirilmasi

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 distribution of intestinal parasites detected in The Dokuz Eylul University Medical Faculty Hospital between 2005 and 2008.]. / Dokuz Eylül Universitesi Tip Fakültesi Arastirma ve Uygulama Hastanesi'nde 2005-2008 Yillari Arasinda Saptanan Bagirsak Parazitlerinin Dagilimi.

A retrospective evaluation of the data from 14,246 patients with gastrointestinal complaints who presented at the parasitology laboratory of the Dokuz Eylul University Medical Faculty Hospital between January 2005 and December 2008 was carried out. Fecal samples of all patients were examined using native-Lugol and the trichrome and Kinyoun acid-fast staining method after sedimentation in fecal concentration tubes. One or more parasites were detected in 1320 (9.3%) of the patients. The distribution of the intestinal parasites was as follows: Blastocystis hominis, 689 (4.83%); nonpathogenic amoebas, 108 (21.82%); Giardia intestinalis, 320 (2.24%); Enterobius vermicularis, 23 (0.16%); Entamoeba histolytica/Entamoeba dispar, 34 (0.24%); and other rare parasites, 78 (0.54%). The results of this study emphasize the fact that intestinal parasitic infections are still an important public health problem.

[Pneumocystis pneumonia in a renal transplant recipient]. / Böbrek transplantasyonlu bir hastada Pneumocystis pnömonisi.

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.

In vitro effects of resveratrol on Trichinella spiralis.

Resveratrol, a natural phytoalexin found mainly in grapes, possesses antibacterial, antifungal, and antiviral properties. However, there is no information about its effects on helminths such as Trichinella sp. In the present study, we investigated the effects of resveratrol on the viability of Trichinella spiralis life stages in vitro. Adult forms, newborn larvae (NBL), and muscle larvae (ML) were incubated with resveratrol at concentrations varying from 12.5 to 200 microg/ml. Resveratrol showed significant anthelmintic activity against NBL and adult forms of Trichinella, but not against ML. Our results suggest that resveratrol may be useful as a therapeutic agent to treat trichinellosis in early stages and warrant its further assessment in animal models of disease.

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.

Demographic status and prevalence of intestinal parasitic infections in schoolchildren in Izmir, Turkey.

Thirteen primary schools from nine areas were randomly selected in the shantytown and apartment districts in Izmir. Fecal specimens were evaluated with native-lugol, formalin-ethyl-acetate sedimentation and with trichrome staining for protozoa and helminths and with cellophane tape for Enterobius vermicularis. Of the study group, 33.4% had one or more parasites. The most common parasite was Blastocystis hominis (14.6%) followed by Enterobius vermicularis (10.1%) and Giardia intestinalis (7.8%). When parasitic distribution was evaluated in association with demographic features, a significant relation was found between the income level and parasitic infection prevalence. Multiple parasitic infections were more prevalent in crowded families (either extended or with many children). When parasitic infection prevalences in the 9-10 and 11-12 years of age groups were compared, the probability in the shantytown primary school group was significantly higher than in the apartment group (p<0.05).