Unilateral phthiriasis palpebrarum infestation in a child during occlusion therapy for amblyopia: Case report.

An 8-year-old mentally retarded boy is brought to the hospital because of itching and burning at his right eye for 10 days. He was on full time right eye occlusion therapy for left amblyopia. Slit lamp examination revealed nits and adult lice anchored to the eyelashes in his occluded eye. Eyelashes and all detected lice and nits were mechanically trimmed, and sent for parasitological examination, which confirmed the diagnosis. Upon familial evaluation for additional infestation, the father was also found to have genital phthiriasis pubis and received appropriate treatment. While phthiriasis palpebrarum in children may signify sexual abuse, a detailed investigation by a child psychiatrist was performed and revealed no sign of abuse. Since the infestation was at only on occluded eye, the most possible explanation for the transmission was evaluated as the misusage of the adhesive patch in our case. In conclusion, sexual abuse should be excluded in children with phthiriasis palpebrarum and parents of amblyopic children on occlusion therapy should be warned about the importance of the hygiene of the patching in order to avoid any kind of infection and infestation.

[Demonstration of Cryptosporidium parvum in immune suppressed rats using nested PCR]. / Immun Sistemi Baskilanmis Siçanlarda Cryptosporidium parvum'un Nested PZR Yöntemi ile Gösterilmesi.

OBJECTIVE: In the present study, the aim is to demonstrate Cryptosporidium parvum 18S small-subunit rRNA gene, in lung and stool samples of immune suppressed rats. This gene region is specific for Cryptosporidium spp. and thus can be used in humans for routine diagnostic procedures. METHODS: Three groups (n=4) of Rattus norvegicus rats were used. The first and second groups were administered dexamethasone, subcutaneously and orally, respectively, for 12 weeks. Rats in the control group were not immune suppressed. Lung and stool specimens were obtained from rats at the end of 12 < sup > th < /sup > week and examined for the presence of C. parvum DNA using Nested PCR. RESULTS: C. parvum DNA was demonstrated in lung and stool samples of rats which were immune suppressed by oral dexamethasone. On the other hand, C. parvum DNA was demonstrated only in stool specimens of the rats which were immune suppressed by subcutaneous dexamethasone. No band pattern was observed in the specimens of the control group. CONCLUSION: The results of the study showed that oral dexamethasone administration was more efficient in generating disseminated cryptosporidiosis in rats compared to subcutaneous dexamethasone administration. In addition, Nested PCR targeting 18S small-subunit rRNA gene can be used to detect Cryptosporidium spp. in respiratory and stool specimens of animals and humans.

Isolation of Toxoplasma gondii strains similar to Africa 1 genotype in Turkey.

INTRODUCTION: Toxoplasma gondii is a protozoon parasite that has a worldwide dissemination. It can cause serious clinical problems such as congenital toxoplasmosis, retinochoroiditis, and encephalitis. Currently, T. gondii genotypes are being associated with these clinical presentations which may help clinicians design their treatment strategy. CASE REPORTS: Two T. gondii strains named Ankara and Ege-1 were isolated from newborns with congenital toxoplasmosis in Central and Western Anatolia, respectively. Ankara and Ege-1 strains were isolated from the cerebrospinal fluid of newborns. According to microsatellite analysis, Ankara and Ege-1 strains were sorted as Africa 1 genotype. CONCLUSION: T. gondii strains isolated in Turkey were first time genotyped in this study. Africa 1 genotype has previously been isolated in immunosuppressed patients originating from sub-Saharan Africa. The reason of detecting a strain mainly detected in Africa can be associated with Turkey's specific geographical location. Turkey is like a bridge between Asia, Europe and Africa. Historically, Anatolia was on the Silk Road and other trading routes that ended in Europe. Thus, detecting Africa 1 strain in Anatolia can be anticipated. Consequently, strains detected mainly in Europe and Asia may also be detected in Anatolia and vice versa. Therefore, further studies are required to isolate more strains from Turkey.

Detection of Pneumocystis in the nasal swabs of immune-suppressed rats by use of PCR and microscopy.

BACKGROUND: Detection of Pneumocystis jiroveci colonization in lungs or oral samples due to high sensitivity of PCR methods results in undue treatment of patients without any symptoms of Pneumocystis pneumonia. The aim of the present study is to demonstrate Pneumocystis carinii in rats, immune suppressed by oral and subcutaneous administration of dexamethasone. MATERIAL/METHODS: Blood, oral, nasal and eye swabs were collected prior to immune suppression and 2, 6, 12 weeks after administration of dexamethasone. Also, samples were collected from lung, heart, liver, kidney, diaphragm, brain, spleen, tongue, muscle, eye, intestine, and feces. Cysts and trophozoites were investigated in stained slides and MSG gene was detected by PCR. RESULTS: The results showed that weight loss is significantly higher in rats administered oral dexamethasone (P<0.05). Microscopy was positive only in lungs of rats orally administered dexamethasone. PCR was positive in lungs and oral swabs of rats prior to the administration of dexamethasone. After the administration of dexamethasone, the MSG gene was detected in oral swabs, lungs, spleen, kidney and (for the first time) in nasal swabs. PCR was positive in nasal swabs during the second and sixth weeks of oral and subcutaneous administration of dexamethasone, respectively. CONCLUSIONS: Presence of P. jiroveci in nasopharyngeal aspirate, oropharyngeal wash, oral swab, induced sputum or BAL, and absence in nasal swab in a patient without symptoms of PCP may support clinician's decision regarding colonization. Overall, detection of P. carinii in nasal swabs of rats by PCR demonstrated that nasal sampling can be used for the diagnosis of Pneumocystis pneumonia.

Preparation of (131)I-Pyrimethamine and evaluation for scintigraphy of experimentally Toxoplasma gondii-infected rats.

We aimed to assess the ability of (131)I-Pyrimethamine scintigraphy to detect the lesions of Toxoplasma gondii infection. An experimental model of toxoplasmosis was developed. The presence of toxoplasmosis was confirmed 60 days after implantation. Pyrimethamine was radioiodinated with I-131. The radioligand was validated by the requisite quality control tests to check its radiolabeling efficiency, in vitro stability and radiochemical purity etc. (131)I-Pyrimethamine (specific activity: 7.08 MBq/µmol) was injected intravenously into the tail vein of the control and infected rats. Static whole body images of the rats were acquired under the gamma camera at 5 min, 45 min, 2 h, 6 h, and 24 h following the intravenous administration of the radioactivity (3.7 MBq/rat). Then the scintigraphic data were analyzed both visually and semiquantitatively. Regions of interest (ROIs) were drawn over the organs (thyroid, stomach, liver, bladder, and soft tissues) to calculate the ratios of the radiotracer in infected vs. control rats. The mean ratio of radiotracer in infected/control rats in the liver and diaphragm was over 1 at 45 min which persisted till 24 h. In conclusion, (131)I-Pyrimethamine may be useful agent for diagnosis toxoplasmosis especially involving liver and diaphragm, needs further preclinical validation before being extended for use in clinical applications.

Action plan to regain unnecessary deferred blood donors due to malaria risk in Turkey.

Malaria was expected to be a major problem during blood donation in Turkey due to existence of malaria cases in southeastern region of Turkey. The present study aimed for the first time, to investigate malaria in "donors deferred for malaria risk" and to determine the regional rates of malaria deferral in Turkey. Blood samples were collected from several Blood Banks of southeastern provinces where local malaria cases still exist and from Blood Bank of Ege University Medical School (EUMS) located in western Turkey where malaria is eradicated decades ago. Plasmodium spp. and specific antibodies were investigated by stained smears, antigen detection, PCR and ELISA. Among the donors deferred for malaria risk, Plasmodium spp. were not detected by microscopy, PCR or antigen detection. Seroprevalances were 2% and 3.92% in western and southeastern regions, respectively. Rate of donor deferral for malaria risk was 0.9% in EUMS and deferrals were exclusively because of travel to southeastern Turkey. In southeastern provinces, deferrals were mainly due to malaria like fever history. The present study first time assessed regional rates of donor deferral due to malaria risk in Turkey. Previously, malaria was expected to be a major problem during blood donation in Turkey due to existence of malaria cases in southeastern region of Turkey. The results of the study showed that 97% of the deferrals were unnecessary. In conclusion, to reduce unnecessary donor deferrals in Turkey, in addition to comprehensive questioning for malaria history, the usage of a malaria antibody screening method should be initiated prior to deferral decision.

Toxoplasma gondii RH Ankara: production of evolving tachyzoites using a novel cell culture method.

Toxoplasma gondii is one of the most researched parasite due to its easy growth both in vitro and in vivo. Tachyzoites, derived from mouse or rat peritoneum encounters ethical and economical problems when used for research or diagnostic purposes. Currently, research has focused on determining the most suitable cell culture environment to reach highest amount of viable tachyzoites with least host cell contamination. However, gene expression changes that take place throughout the adaptation of evolving T. gondii strains to continuous cell cultures appear as a problem. The present study aimed to determine a novel cell culture strategy for T. gondii RH Ankara strain tachyzoites to harvest abundant tachyzoites with least host cell contamination and minimal antigenic variation at predetermined dates to use as an antigen source in serological assays that will facilitate reduction in animal use. To achieve this purpose, T. gondii RH Ankara strain tachyzoites were incubated with HeLa cell at different ratios for two or three days. In all flasks incubated for two days, viability rate reached to 100% and HeLa cell contamination decreased to levels between 0.12-0.5×10(6)/ml. In the flasks with HeLa-tachyzoite ratio 1/8, the tachyzoite yield and viability ratio were 3×10(6)/ml and 100%, respectively, with accompanying 10 fold decrease (0.12×10(6)/ml) in HeLa contamination. During continuous production, highest tachyzoite yield was obtained from the first passage (3.55×10(6)/ml) and until the end of third subculture viability rates and HeLa cell contaminations were between 98.2-99.4% and 0.31-0.37×10(6)/ml, respectively. ELISA, IFA and Western blot analyses showed that the quality, specificity and sensitivity of the antigen harvested from the first passage of cell culture performed at two days intervals were comparable to the antigen harvested from mice and decreased in the following subcultures. Overall, these results demonstrated that T. gondii RH Ankara strain is still evolving to adapt to cell culture environment and therefore such strains continuously produced in cell cultures should be avoided for serological assays. However, the two day short interval cell culture method described herein offers a chance to reduce the animal use intended for the preparation of serological assays' antigen from local evolving strains.

Mesenteric Doppler ultrasonography findings of Echinococcus multilocularis infection: an experimental study.

The diagnosis of alveolar echinococcosis, caused by the larval stage of Echinococcus multilocularis, is often difficult and almost always possible only in the later stages of the disease. In this study, we aimed to evaluate the findings of Doppler ultrasonography images for the diagnosis of intraabdominal non-visceral alveolar echinococcosis in Meriones unguiculatus. Six infected animals were studied. Abdominal Doppler ultrasonography examinations were performed 20-25 days after the implantation. Then, animals were sacrificed and infected specimens were histopathologically examined. The sonographic examinations of the infected animals revealed lobulated, heterogeneous cystic intraabdominal masses. There were echogenic solid areas with hypoechoic and anechoic cystic areas within the lesions. Doppler ultrasound examination revealed vascular islands within those heterogeneous cystic lesions. Color mode showed multiple vascular coding areas within the solid part of the lesions, most of them were venous structures but there were also arterial vessels showing dominant flow with low resistive indices. Vascular structures were supposed to be the invaded native mesenteric vessels within the infiltrating inflammatory mass lesions. We consider that the Doppler ultrasound finding of irregular mesenteric vascular structures within intraabdominal heterogenous mass lesions may be an important sign for the diagnosis of experimental non-visceral alveolar echinococcosis in Meriones unguiculatus.

Comparison of the effects of Artemisia vulgaris and Artemisia absinthium growing in western Anatolia against trichinellosis (Trichinella spiralis) in rats.

Trichinellosis often causing diarrhea and more rarely fever, periorbital edema and myositis in human, is commonly treated with benzimidazole derivatives. The Artemisia genus has been found to be effective against a variety of parasites. In the present study, the efficacy against trichinellosis (Trichinella spiralis) of Artemisia vulgaris and Artemisia absinthium was examined for the first time in rats. The results of trichinoscopy and artificial digestion, during the enteral (adult) phase of the illness show that 300 mg/kg doses of methanol extracts of the aerial parts of A. vulgaris and A. absinthium reduced the larval rate by 75.6% and 63.5% in tongue, 53.4% and 37.7% in diaphragm, 67.8% and 46.2% in quadriceps, and 66.7% and 60.5% in biceps-triceps muscles of rats, respectively. Furthermore, during the parenteral (encapsulated larvae) phase, 600 mg/kg doses of A. vulgaris and A. absinthium extracts decreased the larval rate by 66.4% and 59.9% in tongue, 57.4% and 50.0% in diaphragm, 47.6% and 43.7% in quadriceps, 60.2% and 46.4% in biceps-triceps muscles of rats, respectively. Analysis of antibody also showed that A. vulgaris significantly reduced the antibody response (P<0.05) during the enteral and parenteral phases. Thus, the results of the present study revealed that A. vulgaris could be an alternative drug against trichinellosis.

A case of myiasis in a patient with psoriasis from Turkey.

Myiasis infestations caused by the larvae of flies mostly belonging to Cyclorapha suborders are frequently encountered in Turkey, which is located in the subtropical zone. The skin is a common site for myiasis, and the infestations are likely to develop in infected tissues and poorly attended wounds of the skin. The case, a 30-year-old male patient, was diagnosed with psoriasis 18 years ago. He had psoriatic scales on his right big toe and was receiving corticosteroid and immunosuppressive drugs. A total of 11 fly larvae were removed from the infected right first toe of the patient. Structures of the stigmas seen in the cross-sections taken from the final segments of these larvae were examined and determined as Sarcophaga spp. larvae. Patients with infected tissues should be extremely cautious about their wound hygiene and take required fly control measures especially during summer as flies can find a suitable environment for sustenance.

The first report in Turkey of in vivo cultivation in Rattus norvegicus of Echinococcus multilocularis human strain.

The adult form of the small cestode Echinococcus multilocularis is found in carnivorous animals, especially in the fox. This cestode, which is found in the northern hemisphere in the world, is the cause of a generally fatal disease in humans, known as alveolar echinococcosis (AE). The metacestodes of Echinococcus multilocularis can be experimentally developed in the Meriones unguiculatus species of rodents, and it is possible to use these metacestodes for numerous purposes, primarily for supplying the antigens required for serological diagnosis of the disease. It is with this aim that in this study for the first time in Turkey, Echinococcus multilocularis metacestodes were developed using different kind of rodent, Rattus norvegicus, and an indigenous strain was isolated using the surgical material of a patient diagnosed with alveolar echinococcosis.

The prevalence of cryptosporidiosis in Turkish children, and genotyping of isolates by nested polymerase chain reaction-restriction fragment length polymorphism.

OBJECTIVE: To verify the incidence of cryptosporidiosis among Turkish elementary school students. METHODS: This study was conducted in the Department of Parasitology, Faculty of Medicine, Ege University, Turkey during a 3-month period in 2006. We assessed the fecal samples of 707 children using modified acid-fast and phenol-auramine staining followed by modified Ritchie concentration method. All Cryptosporidium species isolates were analyzed by nested polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) to differentiate the genotypes of the isolates. After the coprological examination, 4 samples were found to be positive for Cryptosporidium species oocysts. RESULTS: In the present study, all 4 oocysts were of zoonotic origin and belonged to Cryptosporidium parvum genotype 2 indicating that in Turkey the potential sources of human cryptosporidiosis is from animals. CONCLUSION: The application of genotyping to clinical isolates of Cryptosporidium has significantly increased our knowledge and understanding of the distribution and epidemiology of this parasite. The PCR and RFLP techniques represent a more rapid and simple method of genotyping to support epidemiological and clinical investigations than conventional analytical DNA techniques.

Incidence of cyclosporiasis in patients with gastrointestinal symptoms in western Turkey.

BACKGROUND: This study was designed to investigate the distribution of cyclosporiasis between October 2003 and October 2004 and the relationship between Cyclospora infection and seasonal as well as patient factors in western Turkey. MATERIAL/METHODS: Stool samples from 4660 immunocompetent patients with gastrointestinal symptoms and 326 immunocompetent patients with allergic symptoms from western Turkey were examined between October 2003 and October 2004 using wet preparation, formalin-ethyl acetate concentration, Trichrome stain, and modified Kinyoun's acid-fast staining methods. RESULTS: Twenty-three patients were found to be infected with Cyclospora oocysts. Parasites such as Cryptosporidium, Giardia intestinalis, Entamoeba histolytica/dispar, Blastocystis hominis and others were also observed. The incidence of cyclosporiasis was higher in summer and early autumn and most of the Cyclospora-infected patients were without diarrhea. CONCLUSIONS: Clinicians with patients from Turkey and abroad who have intestinal symptoms after visiting the country should be aware that Cyclospora infections could be considered as a possible cause of gastrointestinal symptoms in the absence of diarrhea in immunocompetent patients during the summer period in Turkey.

[A human case of cyclosporiasis after traveling in the subtropics]. / Yurtdisi seyahat hikayesi olan bir cyclosporiasis olgusu.

In this study, Cyclospora oocysts were detected in a 64-year old man who complained of persistent diarrhea, abdominal pain, nausea and vomiting after visiting the Greek Islands in a sailing boat. Round oocysts about 8-9 microm in size with wrinkled walls that varied in color from red to pale pink after staining were found in Kinyoun's modified acid fast stained stool samples. Public health offices and laboratories, general practitioners, and medical microbiology labs should be informed that more attention should be paid to cyclosporiasis causing diarrheal illness and which requires specific screening methods with experienced microscopists in laboratories.

A case of nasomyiasis whose agent was Sarcophaga sp.

Sixteen larvae fell from the nose of a 16-year-old girl, who had been hospitalized in the anesthesia intensive care unit for 4 days because of a traffic accident and had been evaluated as E1M2V1 according to Glaskow Coma Scale. These larvae were examined macroscopically and microscopically and it was determined that they were second stage Sarcophage spp. larvae. There was no lesion in the nose of the patient.

A new technique to avoid losing the strains of Echinococcus multilocularis during passaging.

OBJECTIVE: To examine the survival of Echinococcus multilocularis (E. multilocularis) and the formation of protoscoleces, under refrigerated conditions over a 43-day observation period. METHODS: We conducted this study in Ege University, School of Medicine, from May-December, 2004. We included 4 healthy females and 2 males, 8-12 week old M. unguiculatus species (approximately 60-70 gr) for the study, as they are known to be susceptible to E. multilocularis. In this experimental study, we aimed to define a technique to keep the strains during these passages. RESULTS: We found preserved viability and virulence of E. multilocularis stored in RPMI 1640 plus 10% fetal calf serum at +4 degrees C. We accomplished the infection of the Meriones unguiculatus after in-vivo passaging from that flask, which we kept at +4 degrees C, and this proves the viability of protoscoleces and membranous structures perfectly at the 43rd day. CONCLUSION: We also found that E. multilocularis metacestodes and the formation of protoscoleces keep their viability up to 60 days and virulence up to 43 days under this condition. We recommend storing an extra flask during each passage as a precaution against losing strains.

The first report of Echinococcus multilocularis strain isolation from human in Turkey.

The adult form of the small cestode Echinococcus multilocularis (E. multilocularis) is found in carnivorous animals, especially in the fox. This cestode, which is observed in the northern hemisphere of the world, is the cause of a generally fatal, progressing disease in humans, known as "alveolar echinococcosis" (AE). The metacestodes of E. multilocularis can be experimentally developed in the Meriones unguiculatus species of rodents, and it is possible to use these metacestodes for numerous purposes, primarily for antigens required for diagnosis. It is with this aim that in this study for the first time in Turkey, E. multilocularis metacestodes were developed in Meriones unguiculatus and an indigenous strain was isolated using the surgical material from a patient diagnosed with alveolar echinococcosis.

Use of two sensitive and specific immunoblot markers, em70 and em90, for diagnosis of alveolar echinococcosis.

Antibodies against Echinococcus multilocularis metacestodes were screened by immunoblotting sera from patients with alveolar echinococcosis (n = 39), cystic echinococcosis (n = 109), or other parasitic infections (n = 66) and healthy individuals (n = 32). Two antigens, approximately 70 and 90 kDa, are found to be valuable for confirmatory diagnosis, with a sensitivity and specificity of 100 and 99.51%, respectively.