[Detection of Echinococcus granulosus and Echinococcus multilocularis in cyst samples using a novel single tube multiplex real-time polymerase chain reaction]. / Kist örneklerinde yeni bir tek tüp multipleks gerçek zamanli polimeraz zincir reaksiyonu ile Echinococcus granulosus ve Echinococcus multilocularis'in saptanmasi.

Cystic echinococcosis (CE) and alveolar echinococcosis (AE) caused by Echinococcus granulosus and Echinococcus multilocularis, respectively, are important helminthic diseases worldwide as well as in our country. Epidemiological studies conducted in Turkey showed that the prevalence of CE is 291-585/100.000. It has also been showed that the seroprevalence of AE is 3.5%. For the diagnosis of CE and AE, radiological (ultrasonography, computed tomography, magnetic resonance) and serological methods, in addition to clinical findings, are being used. The definitive diagnosis relies on pathological examination When the hydatid cysts are sterile or does not contain protoscolex, problems may occur during pathological discrimination of E.granulosus and E.multilocularis species. In this study, we aimed to develop a novel multiplex real-time polymerase chain reaction (M-RT-PCR) targeting mitochondrial 12S rRNA gene of E.granulosus and E.multilocularis using Echi S (5'-TTTATGAATATTGTGACCCTGAGAT-3') and Echi A (5'-GGTCTTAACTCAACTCATGGAG-3') primers and three different probes; Anchor Ech (5'-GTTTGCCACCTCGATGTTGACTTAG-fluoroscein-3'), Granulosus (5'-LC640-CTAAGGTTTTGGTGTAGTAATTGATATTTT-phosphate-3') and Multilocularis (5'-LC705-CTGTGATCTTGGTGTAGTAGTTGAGATT-phosphate-3') that will enable the diagnosis of CE and AE in same assay. During M-RTR-PCR, plasmids containing E.granulosus (GenBank: AF297617.1) and E.multilocularis (GenBank: NC_000928.2) mitochondrial 12S rRNA regions were used as positive controls. Cysts samples of patients which were pathologically confirmed to be CE (n: 10) and AE (n: 15) and healthy human DNA samples (n: 25) as negative control as well as DNA samples of 12 different parasites (Taenia saginata, Hymenolepis nana, Trichuris trichiura, Fasciola hepatica, Enterobius vermicularis, Toxoplasma gondii, Pneumocystis jirovecii, Trichomonas vaginalis, Cryptosporidium hominis, Strongyloides stercoralis, Plasmodium falciparum, Plasmodium vivax) were used to develop M-RT-PCR. E.granulosus and E.multilocularis control plasmids were constructed to detect analytic sensitivity of the test using TOPO cloning. Positive control plasmids were diluted to determine analytical sensitivity and specificity by distilled water at 10(6)-10(5)-10(4)-10(3)-10(2)-10(1)-1 plasmid copy of dilution in each reaction. According to the results, analytical sensitivity of the assay for E.granulosus and E.multilocularis was 1 copy plasmid/µl reaction. The non-existence of cross reactivity with 12 different parasites' DNA samples showed the analytical specificity of the assay. Displaying Echinococcus DNA in cyst samples among 25 patients and species discrimination as well as non-existence of cross reactivity with human DNA samples showed that the clinical sensitivity and specificity of the assay were 100%. As a result, the M-RT-PCR developed in the present study provided a sensitive, specific, rapid, and reliable method in the diagnosis of echinococcosis and the discrimination of E.granulosus and E.multilocularis from cyst samples.

Toxoplasmosis and Neuropsychological Effects

Toxoplasma gondii is an intracellular protozoan parasite. Approximately 30% of the global population is infected by T. gondii. In chronically infected individuals, the parasite resides in tissue cysts, especially in the brain. There is a growing interest in the role of parasitologic agents in the causation of neuropsychological disorders. In this review, we have explained the interactions between Toxoplasma and its host, mechanisms, and consequences on neural and psychological diseases.

Acute Toxoplasmosis During Pregnancy: A Hard Call

Toxoplasma gondii is an obligate intracellular parasite that infects all animals, including humans, and causes toxoplasmosis. If toxoplasmosis occurs during pregnancy, it may affect the foetus owing to transplacental transmission. Such transmission may lead to foetal complications, some of which can be very serious, e.g. hydrocephaly and chorioretinitis; however, not all cases of acute toxoplasmosis during pregnancy result in foetal complications. The decision whether to continue or terminate the pregnancy is a difficult problem for families as well as healthcare professionals, thus making it important. Here we present a case of acute toxoplasmosis at 6 weeks of pregnancy. The patient was directly advised to terminate the pregnancy. However, with detailed laboratory analyses, close follow-up and treatment to prevent transplacental transmission, she successfully completed the pregnancy and eventually delivered a healthy baby. By presenting this case, we aimed to review acute toxoplasmosis during pregnancy.

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.

Use of the ELISA (Em2-Em18) and Western Blotting Methods on Diagnosis of Alveolar Echinococcosis

Objective: Alveolar echinococcosis (AE) is one of the most lethal parasitic zoonoses in the Northern Hemisphere, and early serological detection is important to start treatment and to improve survival. A total of 50 sera samples of patients diagnosed as having various diseases were examined for by two different serological diagnostic methods. Methods: Em2-Em18 ELISA (Bordier Affinity Products, Crissier, Switzerland) and Echinococcus Western Blot immünoglobulin G (IgG) (LDBIO Diagnostics, Lyon, France) were used for analyisis. Results: A high titer of antibodies was found in 9 of 10 patients diagnosed as having AE with Em2-Em18 ELISA, in 2 of 21 patients with cystic echinococcosis, in 1 of 2 patients with fascioliasis and in 1 patient with chronic hepatitis. The Echinococcus Western Blot IgG test, used as a confirmatory test, showed IgG antibody in 85.7% (18/21) of patients with CE, while all serum samples of 10 patients with AE were evaluated as positive. This method yielded an incorrect diagnosis in the patient with chronic hepatitis and in the patient with granulomatous inflammation with caseification. Samples taken from patients with liver-related diseases and other parasitic-related diseases were found to be negative. Conclusion: The serological methods used in the study were found to be important in the early diagnosis of alveolar echinococcosis in the endemic areas, since it could be used in sero-epidemiological studies.

Does vaginal douching affect the risk of vaginal infections in pregnant women?

OBJECTIVE: To evaluate the relationship between vaginal douching and vaginal infections among women in early pregnancy. METHODS: We conducted this study in the Department of Gynecology and Obstetrics, Family Planning Center, Dr. E. Hayri Ustundag Hospital, Izmir, Turkey, between March 2003 and December 2004. We examined the vaginal swabs of 129 women, asking for termination of pregnancy in a family-planning center as both wet-preparations and cultures for vaginal microorganisms, and recorded the informations on women's vaginal douching habit. RESULTS: Among 129 women examined, 80 had at least one type of vaginal microorganisms. Of 67 vaginal douche users, 48 (71.6%) had at least one type of vaginal organisms, whereas of 62 nonusers, only 32 (51.6%) had microorganism, although age, educational status, coital frequency, age at the first intercourse were not statistically different between the vaginal douche-users and non-users. Especially, Group B Streptococcus (GBS), Enterococcus spp. and Candida spp. were found more frequent in vaginal douche-user women. CONCLUSION: We found that vaginal douching tends pregnant women to genital tract the incidence of vaginal infections, especially those caused by Enterococcus spp. and GBS. As such infections may render such women to high risk in terms of perinatal mortality and morbidity, it would be appropriate to discourage vaginal douching in pregnant women.

[Primary Pelvic Cystic Echinococcosis]. / Primer Pelvik Kistik Ekinokokkoz .

Cystic echinococcosis caused by Echinococcus granulosus is still an important health problem in endemic areas. Cystic echinococcosis may involve different organs or areas with the most common sites being the liver and the lungs. Pelvic involvement has previously been reported and was mainly accepted as secondary to cystic echinococcosis in other organs, isolated pelvic involvement is very rare. In this case report, we aimed to present the case with pelvic cystic mass that was finally diagnosed with isolated pelvic cystic echinococcosis in and after the operation, and we would like to draw attention to include "cystic echinococcosis" in the differential diagnosis of pelvic masses.

Evaluation of (131)I-pentamidine for scintigraphy of experimentally Leishmania tropica-infected hamsters.

We aimed to assess the ability of (131)I-Pentamidine scintigraphy to detect the lesions of Leishmania tropica infection. An experimental model of cutaneous leishmaniasis was developed. The presence of cutaneous leishmaniasis was confirmed. Pentamidine was radioiodinated with (131)I. The radiolabeled pentamidine was validated by the requisite quality control tests to check its radiolabeling efficiency, in vitro stability. (131)I-Pentamidine (activity: 18.5 MBq/100 µl) was injected intracardiacally into infected hamsters. Static whole body images of the hamsters were acquired under the gamma camera at 5 and 30 min, 2, 6 and 24 h following the administration. On the scintigrams, anatomically adjusted regions of interest (ROIs) were drawn over the right feet (target) and left feet (not-target) and various organs. Accumulation of (131)I-Pentamidine at sites of infection is expressed as the target to non-target (T/NT) ratio. The results T/NT ratio decreased with time. In concluding the (131)I-Pentamidine has poor sensitivity in detection of L. tropica infection.

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.

Isospora belli in a patient with liver transplantation.

Isospora belli is an opportunistic protozoon which should be monitored in patients with gastrointestinal complaints such as abdominal pain, nausea and diarrhoea, in both immune-compromised and immune-competent patients. Our case was a 35 year-old male patient who had received a liver transplant because of cirrhosis and hepatic fibrosis. A diarrhoeic stool sample of the patient was sent to the laboratory for microbiological and parasitological analyses. Faecal occult blood was positive and bacteriological analysis was negative. Isospora belli infection was diagnosed by detection of the oocysts in stool samples. Per oral trimethoprim-sulphamethoxazole treatment was given in 500 mg bid dose for 10 days. At the end of the treatment, no oocyst of Isospora belli was seen but non-pathogenic cysts of Entamoeba coli and vacuolar forms of Blastocystis hominis were observed. Two months later the patient had abdominal pain, fatigue and diarrhoea again and parasitological re-evaluation showed oocysts of Isospora belli.

[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 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.

[Contribution to one world, one health: a dog with demodicosis.]. / Tek Tip, Tek Saglik Konseptine Katki: Demodicosisli Bir Köpek.

Dogs are the most preferred pet animal in the world. Canine demodicosis is a skin disease of dogs in which there is proliferation of Demodex canis, an acarine parasite of canine hair follicles, and is typically manifested by alopecia as well as inflammation of hair follicles and sebaceous glands. Secondary bacterial infection often induces pustule and a crusting dermatitis. Two years ago, a police dog eight years old, without any previous health problem, was brought to a private veterinary clinic for edematous and inflammatory lesions on the soles of its feet. In the clinic, antibacterial and antimicotics were applied for treatment of the lesions, but ten months after completion of the therapy the lesions relapsed and the treatment was repeated. But again six months after the last treatment, the lesions spread widely and the general health status of the dog began to worsen. Finally the dog was brought for treatment to the Izmir Bornova Veterinary Research Institution. Microscopic examination of all the skin scrapings revealed the presence of 10-15 adult Demodex mites per cm(2) and the diagnosis was pododemodicosis. Treatment was performed with ivermectin, antibacterial drugs and beta-glucan. The density of Demodex was reduced after two months of therapy and there was clinical and microscopical improvement. Six months after completion of the therapy the lesions disappeared completely.

[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.

Comparison of efficacy of ivermectin and doramectin against mange mite (Sarcoptes scabiei) in naturally infested rabbits in Turkey.

The authors used 14 New Zealand rabbits (5 naturally infested rabbits and 9 in-contact rabbits) for Sarcoptes scabiei treatment in this study. Signs, such as itchy ears, eyes, tail and abdominal skin, alopecia and pyoderma, were considered to be the cause of these disorders. Infested rabbits were grouped according to the intensity of S. scabiei infestation (low, medium and high). Each group was then divided into two subgroups; in one subgroup the rabbits received ivermectin (1%) and, in the other, doramectin (1%). All subgroups received a subcutaneous injection at a dosage of 400 microg/kg body weight every 80 h on three occasions. On day 28 after commencing the treatment, all the rabbits in the first two groups had recovered completely. Although both drugs were applied at the same time and at the same dose, the third group (high degree of infestation), revealed, both microscopically and macroscopically, that ivermectin has more rapid effect than doramectin. Treatment was effective in all groups.

[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.

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.

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.

[The investigation of frequency of cystic echinococcosis in the autopsies committed in the Speciality Department of Istanbul Forensic Medicine Institute]. / Istanbul Adli Tip Kurumu Morg Ihtisas Dairesi'nde Yapilan Otopsilerde Kistik Ekinokokkozis Görülme Sikliginin Arastirilmasi.

Cystic echinococcosis, which is caused by a parasite called Echinococcus granulosus, is a disease frequently find in the liver and in the lungs. In many countries, it is an important public-health problem. This study was done to find out the prevalence of Cystic echinococcosis in forensic autopsies performed for sudden death cases. This investigation was realized between July 2003-January 2004 at Istanbul Forensic Medicine Institution. During this six month period, total of 1687 autopsies were performed. Of those, 56 autopsy cases had macroscopic unknown cysts. The sera obtained from these 56 autopsy cases were examined by IgG Enzyme-linked Immunosorbent assay (ELISA) and IgG Indirect Hemaglutination (IHA) methods. In the end of the serologic evaluation, of 56 cases with macroscopic unknown etiology cysts, 34 cases (60.7%) were serologically positive for cystic echinococcosis, of those 8 were females (14.2%) and 26 were males (46.4%). This study shows that cyst echinococcosis may be find frequently in postmortem cases. The fact that the people who perform autopsies must work carefully and in hygienic conditions is another important lesson obtained from the study.

[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.