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]. / 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.

[Investigation of the cases presenting to Dokuz Eylül University Hospital with tick bites]. / Dokuz Eylül Üniversitesi Hastanesi'ne Kene Tutmasi Yakinmasi ile Basvuran Olgularin Arastirilmasi.

OBJECTIVE: Determination of the properties of ticks, which are the vectors of many infectious diseases and the patients with tick bites are important for the prevention of these diseases. For tist reason, the purpose of this study is to determine the important properties of the cases presenting with tick bites to Dokuz Eylul University Hospital and of the removed ticks from the cases. METHODS: Two hundred seventy three of 294 patients, who presented with tick bites to Dokuz Eylül University Hospital, Izmir, were included in the study. Demographic parameters, symptoms related to tick borne diseases of the patients and the species and other characteristics of ticks removed from humans, also the tick population in the related habitat were investigated. RESULTS: Removed ticks were classified into five genera. The overwhelmingly dominant genera were Hyalomma and it comprised 52.4% of the collection. The majority of these ticks were nymphs. The majority (11.4%) of removed adult ticks were Rhipicephalus sanguineus (R. sanguineus). In most cases (60.7%) the ticks were removed from the patients by medical staff. CONCLUSION: There was no significant difference in removing ticks without damage between the health personnel and the patient or relatives (p=0.133).

[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.

Evaluation of two ELISA and two indirect hemagglutination tests for serodiagnosis of pulmonary hydatid disease.

To establish a definite diagnosis for pulmonary hydatid disease, combination of radiology and serology is useful. In this study, 19 preoperative sera from patients with surgically confirmed pulmonary hydatidosis, 40 sera from patients with other parasitosis and pulmonary diseases, and 20 sera from healthy donors were evaluated using 4 different serological tests, i.e., the commercial ELISA (ELISA-kit) test, the ELISA (ELISA-lab) test prepared in our laboratory, the commercial indirect hemagglutination assay kit (IHA-kit) test, and the IHA test using sensitized sheep red blood cells with tannic acid (IHA-TA). The ELISA-kit was the most sensitive (84.2%) and the most specific test (100.0%). The ELISA-kit also demonstrated the highest positive (100.0%) and negative (95.2%) predictive values. The sensitivity of the ELISA-lab test, that we prepared, was found to be 73.6%, whereas the IHA-kit test and the IHA-TA test were found to be 73.6% and 68.4%, respectively. The specificity of these tests was 96.6%, 98.3%, and 83.3%, respectively. When all 4 tests were assessed together, it was found that the sensitivity had risen to 94.7%. When the ELISA-kit was assessed with the IHA-kit and IHA-TA together, it was found that the sensitivity was 89.5% and 84.2%, respectively. Likewise, the combination of the ELISA-lab and IHA-kit or IHA-TA allowed us to achieve a sensitivity of 84.2% in cases of pulmonary echinococcosis. In conclusion, the diagnosis would be imminent if least 2 tests were applied together.

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.

Diagnosis and treatment of demodectic blepharitis.

The aim of this study was to investigate the prevalence of Demodex spp. in the eyelash follicles obtained from patients seen in our ophthalmology clinic, to define the symptoms of this infestation, and to examine the effectivity of the therapy. This study was conducted in Department of Ophthalmology and Parasitology, Dokuz Eylül University, School of Medicine. Our study included 82 cases that were seen in the Ophthalmology Department and Parasitology Department for various reasons. We have also observed that the presence of Demodex spp. provokes itching and redness in the eyes and that using baby shampoo for cleansing the face reduces the risk of infestation. After the treatment of 32 cases with 4% pilocarpin HCl gel, we achieved a total cure in 12 eyes (37.5%), partial improvement in 13 eyes (40.6%), (making a total of 25 eyes, 78.1%). The treatment was unsuccessful in 7 eyes (21.9%). In patients with Demodex spp. cleansing with baby shampoo and treating by pilocarpin gel may be used in treatment.

[The ability of 67Ga scintigraphy to detect Trichinella spiralis]. / Trichinella spiralis'in Tanisinda Gallium-67 Sintigrafisinin Yeri.

To assess the value of (67) Ga scintigraphy in the lesions of Trichinella spiralis, an animal model was developed. Three infected and five healthy animals were enrolled in this study. After the injection of 7.4 MBq (200 micro Ci) (67) Ga citrate intravenously, static images from whole body were obtained at 24(th), 48 (th) and 72 (nd) hours Visual and semiquantitative analyses were performed. In semiquantitative analysis, activity counts were calculated over the whole body, diaphragm, neck muscles and extremities muscles of all animals. Between the infected animals and control group, mean bodies activity ratios were compared as visually and statistically. (67) Ga scintigraphy can't successfully demonstrate the lesions of Trichinella spiralis.

An outbreak of gastroenteritis associated with intestinal parasites.

Intestinal protozoa are one of the leading causes of waterborne outbreaks. Stool samples of 196 residents from a village of Izmir, using the public water supply, were collected during an outbreak of gastroenteritis. Patients were asked to fill out a questionnaire reporting on gender, age, gastrointestinal symptoms, whether or not there was a toilet in the house, their hygiene practices, and similar symptoms in the household members. Of the patients who had gastrointestinal symptoms (74.5%), diarrhea was observed in 69.5% whereas bloody and mucoid stools were observed in 20.4 %. The stool samples were examined for intestinal parasites by wet mount and trichrome stain and were also cultured in the Robinson medium. Pathogenic parasites were detected in 11 samples (5.6%) as follows: Giardia intestinalis in 7, Hymenolepis nana in 1 and Blastocystis hominis in 4. Entamoeba histolytica/dispar was not detected by direct wet mount in any of 8 patients who had E. histolytica/E. dispar in culture whereas it was detected in the trichrome stained slides of 3 patients. Amoeba prevalence in the 15-44 age-group was significantly high when compared with the 0-14 age group. The prevalence of pathogenic parasites was high among the people who had a toilet outdoors. Drinking water was thought to be a principal source of this outbreak.

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).

Albendazole: single or combination therapy with permethrin against pediculosis capitis.

Pediculosis capitis is a worldwide problem and a growing concern because of resistance to pediculicides. In the present study, we investigated whether albendazole could be used in the treatment of pediculosis capitis in combination with 1% permethrin or alone. A total of 150 children were randomly divided to five groups of 30 each. Group 1 got albendazole in a single dose (400 mg), group 2 got albendazole at 400 mg for 3 days, group 3 was given 1% permethrin, group 4 took 1% permethrin and albendazole in a single dose (400 mg), and group 5 got 1% permethrin and albendazole in a dose of 400 mg for 3 days. Groups given albendazole were also given another 400 mg dose of albendazole after 1 week. The success rate of treatment at the 2-week follow-up for all groups was 61.5%, 66.6%, 80.0%, 84.6%, and 82.1%, respectively. No statistically significant difference was found between the groups. The results of this study suggest that albendazole is effective against pediculosis capitis and there is no synergistic effect between albendazole and 1% permethrin.

[Evaluation of patients presenting with a suspicion of cystic echinococcosis to the serology laboratory of the Parasitology Department of Dokuz Eylül University Medical Faculty]. / Dokuz Eylül Universitesi Tip Fakültesi Parazitoloji Anabilim Dali Seroloji Laboratuvari'na Kistik Ekinokokkozis süphesiyle basvuran hastalarin degerlendirilmesi.

Cystic echinococcosis (CE) is a zoonosis, caused by the metacestode form of Echinococcus granulosus. The diagnosis of CE is difficult using the clinical features of the disease and it depends on the combination of serological methods aimed at determining the specific antibody response and on imaging techniques that support the serology. In this study, 465 patients who presented with a suspicion of CE to the serology laboratory of the Parasitology Department of Dokuz Eylul University between January 2003 and June 2004 were evaluated. The specific anti-E. granulosus antibodies in the serum samples of patients were examined by an in-house enzymelinked immunosorbent assay (ELISA) test and a commercial indirect hemagglutination (IHA) test. Seventy eight (17%) of the patients were positive with ELISA and 65 (14%) of them were positive with IHA with varying titrations. Fifty six (12%) of the sera were positive with both methods. Fifty-six of the eighty patients who underwent CE surgery at different times before this study were found to be seropositive, while the remaining 24 were found to be seronegative. We concluded that it is very important to combine at least two methods in the serological diagnosis of CE.

[In vivo development of Trichinella spiralis in rats]. / Trichinella spiralis'in in vivo siçanlarda olusturulmasi.

The causative agent is Trichinella spiralis which lives in the intestines in adult form or in skeletal muscles in larval form. After ingestion of raw or undercooked infected meat that contains larvae of trichinella, these larvae enter the blood and the lymphatic circulation. They usually settle in the skeletal muscles and develop further. This study was conducted in the Department of Parasitology of the Dokuz Eylul University, School of Medicine. The aim of this experimental study was to develop Trichinella spiralis in vivo experimentally using Wistar albino rats. It was also found that Trichinella spiralis can be experimentally developed in the Rattus norvegicus species of rodents, and it is possible to use the obtained Trichinella spiralis larva's for numerous purposes, in particular for supplying the antigens required for serological diagnosis of the disease. In addition, we investigated the range of the the skeletal muscles in which the causative agent, Trichinella spiralis, lives in larval form.

[Evaluation of three different ELISA kits for the diagnosis of trichinellosis]. / Trisinellozis tanisinda uç farkli ELISA kitinin degerlendirilmesi.

Human trichinellosis caused by Trichinella spp. is a widely distributed parasitic disease, acquired by ingestion of undercooked or raw meat containing larvae of the parasite. In January 2004, a trichinellosis outbreak following the consumption of raw minced meat ball, occurred in Izmir, Turkey. In the present study, an in-house excretory/secretory (ES) IgG ELISA and two commercial IgG ELISA kits (Kit # I; IBL, Hamburg, kit # II; Cypress, Belgium) have been used to evaluate their diagnostic efficiencies in this outbreak. Serum samples were collected from 100 patients with trichinellosis, from 16 patients with other parasitosis and autoimmune diseases (2 toxocariasis, 8 hydatic cyst, 1 fascioliasis, 5 rheumatoid arthritis) and from 20 healthy subjects. While the sensitivity of in-house ELISA has been found as 100%, the sensitivities of commercial kits number I and II have been found as 75% and 48%, respectively. The specificities of these three tests have been detected as 93.7%, 87.5% and 100%, respectively. The results obtained in this study suggest that in-house ELISA is of significant diagnostic value for the diagnosis of trichinellosis.

[Evaluation of IHA, ELISA and Western Blot tests in diagnosis of pulmonary cystic hidatidosis]. / Akciger hidatidozunun tanisinda IHA, ELISA ve Western Blot testlerinin degerlendirilmesi.

Pulmonary cystic hidatidosis caused by the larval stages of Echinococcus granulosus is a common parasitic disease in Turkey and throughout the world. In this study IHA, ELISA and Western Blot (WB) tests were performed with a panel of 59 sera from 31 surgically confirmed pulmonary cystic hidatidosis patients, 18 patients with pulmonary disease other than cystic hidatidosis and 10 healthy individual. The overall sensitivity of the IHA, ELISA and WB tests used for the serodiagnosis of pulmonary cystic hidatidosis were found as 96.7%, 87.1%, 100% and the specificities were 82.2%, 89.2% and %85.7, respectively. Using the WB test 8-12 kDa, 24 kDa and 124 kDa bands were detected as valuable for surgically confirmed patients' sera. One or more of these bands were also detected in sera of four patients with other pulmonary diseases false-positively. In conclusion conventional serologic test like IHA and ELISA is valuable in diagnosis of pulmonary cystic hidatidosis, also evaluation of some specific bands in WB would contribute to the diagnosis.

Prevalence of Demodex in health personnel working in the autopsy room.

The health personnel working in the autopsy rooms and laboratories are considered among the professional groups carrying a high risk of contamination with infection. In this study, we have investigated the prevalence of Demodex, which provides a convenient environment for bacteria and acts as a vector for certain microorganisms, in the health personnel working in autopsy rooms. The study, which was cross-sectional, was conducted on 58 health personnel working in autopsy rooms. A questionnaire was administered to the individuals. Specimens were obtained from 4 different regions of the face using the standard skin-surface biopsy technique. The data obtained were then evaluated statistically. This study group consisted of 76% males and 24% females. The prevalence of Demodex was 26%. Demodex was found to occur more frequently in women. The most frequent occurrence of Demodex was seen among people with darker complexion and in cheek localizations. We believe that Demodex, which has a role in the pathogenesis of dermatologic diseases, provides a convenient environment for bacteria, and acts as a vector for some pathogenic microorganisms, poses a risk as far as individuals working in the autopsy room are concerned.