Subtype distribution and molecular characterization of Blastocystis from hemodialysis patients in Turkey.

INTRODUCTION: The aim of this study was to determine the Blastocystis prevalence and subtypes in hemodialysis patients in Turkey. METHODOLOGY: Eighty-four patients diagnosed with end-stage renal failure who were undergoing hemodialysis and 20 healthy volunteers were enrolled. Blastocystis presence was investigated by native-Lugol, trichrome staining, PCR using STS primers, and DNA sequencing analysis. RESULTS: Among the stool samples from the hemodialysis patients, 9.52% (8/84) were found to be Blastocystis-positive with the native-Lugol and trichrome staining. Seven of the eight Blastocystis isolates were subtyped using STS primers. Blastocystis subtype distribution was as follows: one had ST1, two had ST2, two had ST3, two had ST3+ST6, and one was not subtyped. Blastocystis positivity was detected in two healthy control (2/20, %10), one subject had ST1, and the other was not subtyped. All subtypes identified by PCR were confirmed by the sequencing analysis. In the two samples that had mixed subtypes (ST3+ST6) when using the STS primers, only ST3 was detected in the sequencing analysis. Although some patients have multiple symptoms, the most common symptoms in Blastocystis positive patients were bloating (5/8), diarrhea (4/8), nausea and vomiting (2/8), and gas and weight loss (1/8). Also, only one patient had Giardia intestinalis. CONCLUSIONS: This was the first study to determine the Blastocystis subtypes in hemodialysis patients. A rare subtype, ST6, was identified in two of the patients. Thus, the ST6 infections were attributable to transmission from poultry infections. The presence of this unusual subtype suggests the need for further extensive studies of hemodialysis patients.

Blepharitis Caused by Demodex

Objective: Demodicosis is a Skin disease in humans caused by Demodex folliculorum (D. folliculorum) and Demodex brevis (D.brevis) mites. Demodex infestation is mostly located in sebaceous and meibomian glands in the hair follicles. The aim of this study was to determine the age and sex distribution of Demodex infestation in patients with blepharitis. Methods: Between 2011-2018, eyelashes from 335 patients with blepharitis, visual impairment or conjunctivitis attached to the cellophane tape were sent to the laboratory of Department of Medical Parasitology in Çukurova University Medical Faculty and were examined within 1 hour. Results: D. folliculorum was detected in 143 (42.6%) of the 335 patients. Of the patients in whom D. folliculorum was detected, 50 (35%) were female and 93 (65%) were male. The mean age of patients with D. folliculorum was 64.1 years and the mean age of patients without D. folliculorum was 52.7 years. Conclusion: In patients with blepharitis, the incidence of Demodex increases with age.

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.

The epidemic typhus and trench fever are risk for public health due to increased migration in southeast of Turkey.

Pediculus humanus capitis is a small ectoparasitic insect that has lived and feds on human beings for thousands of years. Molecular techniques have been used for Pediculus species identification and evolutionary, phylogenic, and ecological studies. A total of 23 adults of P. h. capitis were collected in Gaziantep, located in southeast Turkey, and DNA was isolated from all P. h. capitis using DNA extraction kit. All DNA samples were screened for investigate of Ricettsia prowazekii, Bartonella quintana and Borrelia recurrentis with real-time polymerase chain reaction. In addition, we investigated genetic variation in DNA samples of Pediculus humanus capitis using the cytochrome oxidase I genetic DNA sequence. We found 4 (17.4%) Ricettsia prowazekii and 3 (13.1%) Bartonella quintana in DNA samples of Pediculus humanus capitis, while we did not find any Bartonella recurrentis in any of the DNA samples. We demonstrated 1.8% genetic variations in DNA samples of Pediculus humanus capitis with Bartonella quintana. The phylogenetic tree based on the cytochrome oxidase I gene revealed that P. h. capitis in southeast Turkey are classified into two clades (clade A, clade B) and Bartonella quintana was found in only clade B. However, we did not find any genetic variations in other DNA samples in this region. The genetic variations may be related to P. h.capitis vector of Bartonella quintana has found in this study. In addition, this study was shown that P. h. capitis do transmit Rickettsia prowazekii and Bartonella quintana to people, epidemic typhus and trench fever may emergence in Gaziantep southeast of Turkey in the future.

Subtype analysis of Blastocystis isolates using SSU rRNA-DNA sequencing in rural and urban population in southern Turkey.

Blastocystis is a common and emerging parasite often seen in many studies conducted in urban population, with scanty reports on rural communities. However, little is known about the public health significance of Blastocystis infection. A total of 28 Blastocystis isolates from 17 (17/28, 60.71%) patients living in rural area and 11 (11/28, 39.29%) patients living in urban area were screened with seven kinds of sequenced-tagged site primers for identification of subtype. PCR products were sequenced with same combination of primers using the BigDye Terminator V 3.1 cycle sequencing kit, as per the manufacturers' protocol on the 3730 DNA analyzer (Applied Biosystems, Carlsbad CA, USA). The cross-comparison of the Blastocystis sequences of samples were determined by the neighbor-joining method based on a distance matrix between sequence pairs to generate dendograms. The following subtypes were identified; subtype 1 (10/28, 35.7%), subtype 3 (7/28, 25.0%), subtype 2 (5/28, 17.8%), subtype 4 (3/28, 10.7%), subtype 5 (1/28, 3.6%), subtype 6 (1/28, 3.6%), and subtype 7 (1/28, 3.6%) in all DNA samples. The comparison of Blastocystis subtypes distribution among the patients from rural and urban area revealed subtype 5 (1/17, 5.9%), subtype 6 (1/17, 5.9%) and subtype 7 (1/17, 5.9%) from patients of rural area but not any of these subtypes in patients living urban area. This study is the first large-scale study to examine the occurrence of Blastocystis in Turkey to shed lights on the cosmopolitan distribution of Blastocystis subtypes in southern part of Turkey. Subtype 5, subtype 6 and subtype 7 were determined in only rural area. The findings of this study suggest that Blastocystis is transmitted from animal to human and possess a zoonotic potential.

Pediculosis capitis is a growing neglected infestation due to migration in southeast Turkey.

Demographic, socio-economical, and environmental changes affecting prevalence of Pediculosis capitis. The aim of this study was to investigate the prevalence of P. capitis and external factors affecting the distribution of P. capitis. A total of 6004 primary-school students between 5 and 11 years were screened for P. capitis at 28 different primary-schools in Gaziantep, located in southeastern of Turkey, during different two education terms (First education term is in September 2013 to May 2014, second education term is in September 2014 and May 2015). The prevalence of P. capitis was found to be positive 1.5 % (90/6004) and 6.9 % (415/6004) in first education term and in second education term, respectively. In this study shown that the rate of P. capitis's prevalence was increased 5.4 % in Gaziantep. P. capitis is a neglected infestation and it has re-emerged in Gaziantep, located in the southeastern of Turkey. Health staff member must improve health education programs in primary-school students especially girl students.

The role of domestic tap water on Acanthamoeba keratitis in non-contact lens wearers and validation of laboratory methods.

Acanthamoeba is increasingly recognized as an important cause of keratitis in non-contact lens wearers while contact lens wear is the leading risk factor for Acanthamoeba keratitis (AK). It is unlikely that the Acanthamoeba colonization is a feature which is effective only in patient's homes with infectious keratitis since the organism has been isolated from domestic tap water. Two hundred and thirty-one (231) corneal scrapings were taken from infectious keratitis cases, and four contact lens solutions and domestic tap waters were taken from 22 out of 44 AK-diagnosed patient's homes. Microscopic examination, culture, PCR, real-time PCR and DNA sequencing analyses were used for AK-diagnosed samples. The real-time PCR was the most sensitive (100 %) one among the methods used in diagnosis of AK. The 44 (19.0 %) out of 231 corneal scrapings, 4/4 (100 %) contact lens solution and 11/22 (50 %) of domestic tap water samples were found to be positive by real-time PCR for Acanthamoeba. A. griffini (T3), A. castellanii (T4) and A. jacobsi (T15) genotypes were obtained from corneal scrapings, contact lens solutions and domestic tap water samples taken from the patient's homes diagnosed with AK. The isolation of Acanthamoeba containing 6/22 (27.3 %) A. griffini (T3), 14/22 (63.6 %) A. castellanii (T4) and 2/22 (9.1 %) A. jacobsi (T15) from the domestic tap water outlets of 22 of 44 (50 %) of patient's homes revealed that is a significant source of these organisms. A. griffini (T3) and A. jacobsi (T15) genotypes have not been determined from AK cases in Turkey previously. Thus, we conclude that Acanthamoeba keratitis is associated with exposition of patients who has ocular trauma or ocular surface disease to domestic tap water in endemic or potentially endemic countries.

Comparison of clinical samples and methods in chronic cutaneous leishmaniasis.

This study aimed at finding out the most effective clinical samples and methods in chronic cutaneous leishmaniasis (CCL). Smear, aspiration fluid, and filter paper samples were taken from 104 skin lesions of suspected cases with CCL, and they were compared using microscopic examination, culture, and molecular methods. We characterized four different forms of CCL and identified the causative agents in CCL forms using high-resolution melting curve real-time polymerase chain reaction assay. We observed that smear was detected to be the most sensitive (63.5%) among clinical samples, and real-time polymerase chain reaction method was the most sensitive (96.8%) among the methods used in diagnosis of CCL. We identified 68.8% Leishmania tropica and 31.2% L. infantum in papular lesions, 69.2% L. infantum and 30.8% L. tropica in nodular lesions, 57.9% L. tropica and 42.1% L. major in ulcerating plaque lesions, and 55.5% L. tropica and 44.5% L. major in noduloulcerative lesions in CCL patients.

Feconomics®; a new and more convenient method, the routine diagnosis of intestinal parasitic infections.

Direct wet mount examination and concentration are the most commonly used methods for detecting intestinal parasites from fecal samples. Concentration methods are used when there are fewer protozoan cyst, coccidian oocyst, microsporidial spore, helminth egg, and larvae in the fecal samples. Early detection of the causative intestinal parasites plays a significant role in implementing timely and correct treatment, which relieves the patients' symptoms and also prevents recurrences. Formalin-ethyl acetate concentration (FEAC) is believed to be a gold standard method to detect most intestinal parasites. Thus, in this study, we evaluated the diagnostic value of Feconomics® [manufactured by Salubris Inc, Boston, USA. Patent application number (TR): 2010/07549] which is a simple, new, and rapid fecal concentration method for the detection of the intestinal parasites in human beings. We also compared the FEAC with Feconomics® and direct wet mount examination. A total of 918 fecal samples were collected from the patients suspected to have intestinal parasitic infection. Samples were examined with the direct wet mount, FEAC, and Feconomics® methods. Different parasite species 15.9% (146/918) with Feconomics®, 13.3% (122/918) with FEAC, and 9.8% (90/918) with direct wet mount examination, Feconomics® > FEAC > direct wet mount examinations were detected. They were statistically compared considering FEAC as the gold standard for parasitological diagnosis; the sensitivity and specificity of Feconomics® were calculated as 96 and 97%, respectively. Blastocystis hominis was found to be the most common parasite, followed by Giardia lamblia with direct wet mount examination, FEAC, and Feconomics® methods. Feconomics® proved to be better than not only FEAC in concentrating parasite egg and cyst forms as well as in maintaining characteristic morphology but it is also better in direct wet mount examination. Feconomics® eliminates the need for centrifugation by using absorbent beads that help the homogenization and concentration of the sample. Feconomics® in this study was considerably better than FEAC in detecting the trophozoites of Giardia lamblia. We suggest that Feconomics® be used for the routine diagnosis of intestinal parasitic infection in rural areas of developing countries due to the fact that a centrifuge is not required and it eliminates large stool particles.

Clinical characterization of unusual cystic echinococcosis in southern part of Turkey.

BACKGROUND AND OBJECTIVES: The incidence of primary extrahepatic cystic echinococcosis (CE) is rare. Generally, radiological and serological findings can help establish the diagnosis of hepatic and pulmonary CE, but a CE in an unusual location with atypical radiological findings may complicate the differential diagnosis. The objective of this study is to present the characteristics of cases with extrahepatic CE in respect of sites of involvement, clinical presentations, radiological findings, serological diagnostic evaluations, and outcomes of infected patients. DESIGN AND SETTINGS: A retrospective analysis of surgically treated CE was conducted between January 1993 and January 2014 in the General Surgery, Pediatric Surgery, Urology, Cardiovascular Surgery, Neurosurgery, and Orthopedics departments of University of Cukurova, Faculty of Medicine, Balcal Hospital. PATIENTS AND METHODS: Among the 661 patients managed for CE, 134 had unusual sites of involvement. Radiological and serological examinations were used to differentiate CE from alveolar echinococcosis. RESULTS: Of 134 cases with unusual sites of involvement, 32 cases had liver CE (23.9%), 7 cases had lung CE (5.2%), and 2 cases had concomitant liver and lung CE (1.5%). In 93 (69.4%) cases, unusual organ involvement was isolated without any liver or lung involvement. The mean age was 45 years. Abdominal pain was the main symptom and was found in 104 patients. Thirty-one (23.1%) of 134 extrahepatic CE cases were evaluated as negative with indirect hemagglutination (IHA). However, positive results were obtained in 54 cases evaluated with Echinococcus granulosus IgG Western blot (WB), including 10 IHA-negative cases. CONCLUSION: CE with unusual localizations may cause serious problems of diagnostic confusion. The combination of clinical history, radiological findings, and serological test results (especially the WB) are valuable in diagnosing extrahepatic CE.

Investigation of Cryptosporidium spp. antigen by ELISA method in stool specimens obtained from patients with diarrhea.

Cryptosporidium spp. is an important parasitic protozoan causing diarrhea in developing and developed countries. The agent causes severe life-threatening diarrhea especially in immunocompromised hosts. Diagnosis of the Cryptosporidium oocyst in stool samples by conventional microscopy is labor-intensive and time-consuming. Thus, we aimed to evaluate the usefulness of a copro-antigen enzyme-linked immunosorbent assay (ELISA) test in detecting Cryptosporidium spp. from fecal specimens. For this aim, microscopy and specific antigen detection methods were compared to determine Cryptosporidium spp. In addition, specific antigen by ELISA method in stool was investigated in order to find out whether or not it contributes to the diagnosis of Cryptosporidium spp. One hundred and fifty-four stool specimens taken from patients whose ages ranged from 0 to 86 with diarrhea applied to Department of Parasitology, Balcali Hospital of Cukurova University in Adana, Turkey were used. All samples were examined for Cryptosporidium spp. antigen by ELISA and oocysts via gold standard modified acid-fast staining, between October 2008 and July 2009. Eight (5.19%) specimens were found to be positive by modified acid-fast staining method and 37 (24.03%) specimens by copro-antigen ELISA method were found to be positive. The sensitivity and specificity for copro-antigen ELISA were 100% and 80.1%, respectively. The results of copro-antigen ELISA indicate that the simple, rapid, reliable, and standardized immunoassay test is sensitive and specific for routine diagnosis and may be useful for large-scale epidemiological studies of cryptosporidiosis.

Evaluation of the transmission mode of B. hominis by using PCR method.

Blastocystis hominis is a common intestinal parasite observed in fecal examination. On the other hand, the transmission of this parasite is certainly unknown. The transmission of B. hominis can be realized by animal contact and the contamination by water and food with excreted cysts from the reservoir hosts. B. hominis isolated from 25 humans, their pets, and tap water was identified by polymerase chain reaction using sequenced tag site primers in this study. B. hominis isolates obtained from humans and pets were identified as subtype1, subtype2, and subtype3 while B. hominis isolates obtained from tap water were also identified as subtype1. The B. hominis isolates obtained from humans in this study were defined as the same as the subtypes of the B. hominis isolates obtained from the pets, of which these people keep at their homes, and the tap water. These findings reveal that the source of B. hominis infection could be pets and tap water.

Detection of Plasmodium vivax by nested PCR and real-time PCR.

Malaria is endemic in the Cukurova region while it is sporadic in other regions of Turkey. Therefore, the laboratory and clinical diagnosis of malaria is important for the treatment of malaria. In this study, 92 blood samples that were taken from the suspected malaria patients for routine diagnosis in a period of 10 years between 1999 and 2009 were analyzed. All of these blood samples were examined by microscopic examinations using Giemsa-stained thick blood films, nested PCR, and real-time PCR. The sensitivity-specificity and positive-negative predictive values for these diagnostic tests were then calculated. It was found that the positive predictive values of microscopic examination of thick blood films, nested PCR, and real-time PCR were 47.8%, 56.5%, and 60.9% for malaria, respectively. The real-time PCR was found to have a specificity of 75% and sensitivity of 100%, while specificity and sensitivity of nested PCR was found 81.2% and 97.7% according to the microscopic examination of thick blood films, respectively.

Comparison of microscopic examination, rK39, and PCR for visceral leishmaniasis diagnosis in Turkey.

The laboratory diagnosis of visceral leishmaniasis is based on microscopic examination, culture, serological tests, and molecular methods. In this study, we examined 50 blood specimens from suspected visceral leishmaniasis patients by microscopic examination, recombinant antigen dipstick test (rK39), and polymerase chain reaction (PCR) in the University of Cukurova, Faculty of Medicine, Parasitology Department in Turkey. We calculated the sensitivity-specificity and positive-negative predictive values for these diagnostic tests. We found that positive predictive value of microscopy examination, rK39 dipstick test, and PCR were 20%, 24%, and 58% for visceral leishmaniasis, respectively. When we compared polymerase chain reaction, recombinant antigen dipstick test, and microscopic examination for visceral leishmaniasis diagnosis, the polymerase chain reaction is more sensitive (100%) than recombinant antigen dipstick test and microscopy examination.

Identification of Blastocystis hominis isolates from asymptomatic and symptomatic patients by PCR.

Despite years of study, the pathogenic role of Blastocytis hominis is still controversial. Genotypic differences between the asymptomatic and symptomatic isolates should assist in determining the pathogenicity of Blastocystis. In this study, we genotyped 32 Blastocystis isolates obtained from 12 asymptomatic healthy individuals and 20 symptomatic patients pain by polymerase chain reaction using known seven kinds of sequence tagged site primers in this study. When we compared genotype of Blastocystis isolates between the symptomatic and asymptomatic patient group, we found that subtype3 is the most dominant genotype in asymptomatic individual (9/12) and subtype1 determined all of symptomatic patients (20/20).

[Cystic echinococcosis in Turkey from 2001-2005]. / Türkiye'de 2001-2005 Yillari Arasinda Kistik Ekinokokkozis.

Cystic echinococcosis (CE) caused by the metacestode form of Echinococcus granulosus is a major public health problem especially in animal-raising regions of the world. In the present study, CE cases were determined during 2001-2005 by investigating different hospital and health directorship documents and Health Ministry documents, retrospectively. Our results show that there were 2534 (13.13%) cases in the Marmara region; 2114 (16.94%), in the Aegean region; 2578 (16.09%), Mediterranean region; 5404 (38.57%), in the Middle Anatolian region; 428 (5.70%), in the Black Sea region; 844 (6.80%), in the eastern Anatolian region; and 887 (2.75%), in the southeastern Anatolian region making a total of 14,789 CE cases. Finally, it has been determined that the patients were hospitalized for a total of 149,464 days.

Computed tomographic findings of trichuriasis.

In this report, we present computed tomographic findings of colonic trichuriasis. The patient was a 75-year-old man who complained of abdominal pain, and weight loss. Diagnosis was achieved by colonoscopic biopsy. Abdominal computed tomography showed irregular and nodular thickening of the wall of the cecum and ascending colon. Although these findings are nonspecific, they may be one of the findings of trichuriasis. These findings, confirmed by pathologic analysis of the biopsied tissue and Kato-Katz parasitological stool flotation technique, revealed adult Trichuris. To our knowledge, this is the first report of colonic trichuriasis indicated by computed tomography.