Determination of Parasitic Contamination in Vegetables Collected from Local Markets in Izmir Province, Türkiye

OBJECTIVE: Fresh vegetables are an important part of a healthy and nutrient-rich diet but the consumption of raw vegetables without proper washing is the main way for transmission of parasites. This study was aimed at determining the rate of parasitic contamination in prewashed fresh vegetables sold at randomly selected 10 retail markets which is the last step to reach the consumer in Izmir, Türkiye. METHODS: A total of 80 samples selected from eight types of vegetables including tomato, spinach, lettuce, rocket, mint, parsley, dill, and cucumber were examined for parasitic agents microscopically by sedimentation method after washing samples with normal saline. Statistical analysis was performed using SPSS software version 20.0. RESULTS: Protozoan cysts, helminth eggs, and larvae were detected in 21 (26.2%) of 80 samples from eight different vegetable species. Rhabditidiform larvae 18.7%, Blastocystis spp. 5%, Toxocara spp. 2.5%; Ascaris spp., Fasciola spp., Entamoeba histolytica/ Entamoeba dispar and hooked worms were found in 1.2%. Spinach and mint samples were contaminated with parasites significantly more than other fresh vegetable samples (p<0.008, odds ratio =80.0; p<0.017, odds ratio =46.6 respectively). Cruznema spp., a plant nematode, was found at the highest rate according to the results of culture, polymerase chain reaction and sequencing, respectively. CONCLUSION: In this study, the parasitic contamination was found in approximately one of the four vegetables sold in randomly selected markets in Izmir. These findings show that vegetables sold in local markets can cause parasitic infections if they are consumed without adequate washing and awareness should be raised on this issue. In addition, it was concluded that morphological examinations should be confirmed by molecular studies and sequencing as much as possible in order to avoid misdiagnosis of rhabditidiform larvae.

Distribution and Phylogenetic Analysis of Subtypes and Alleles of Blastocystis sp. in the Stool Samples Collected from Patients with Gastrointestinal Complaints in Izmir, Turkey.

PURPOSE: Blastocystis sp. is one of the most prevalent intestinal protozoa found in humans and many other animals. The present study aimed to examine the distribution and genetic diversity of Blastocystis sp. in stool samples from patients with gastrointestinal complaints in Izmir, Turkey. METHODS: All stool samples of 439 patients with gastrointestinal complaints were examined by native-Lugol and trichrome staining. To investigate the presence of Blastocystis sp. in stool samples, DNA was isolated, and PCR was performed with the barcode region in the SSU rRNA gene. PCR positive samples were sequenced to identify subtypes and alleles of Blastocystis sp. RESULTS: The prevalence of Blastocystis sp. was found to be 16.6% (73/439) in patients with gastrointestinal complaints in Izmir, Turkey. Three different Blastocystis sp. subtypes were identified. ST3 (28/55; 51.0%) was the most common subtype followed by ST2 (19/55; 34.5%) and ST1 (8/55; 14.5%). Itching and diarrhea were the most prominent clinical symptoms in Blastocystis sp. positive patients. When clinical symptoms and subtypes were compared, diarrhea was found in 62.5%, 47.4%, and 46.4% of patients with ST1, ST2, and ST3 subtypes, respectively. In addition, itching was found in 37.5%, 32.1%, and 21.1% of patients with ST1, ST3, and ST2, respectively. Six distinct alleles were identified by allele analysis of Blastocystis 18S rRNA gene: allele 4 for ST1, alleles 9, 11, and 12 for ST2, and alleles 34 and 36 for ST3. In this study, Blastocystis sp. was detected in 16 of 21 districts, including the central and rural districts of Izmir. Although ST1 was detected in central districts, it was not found in rural districts. CONCLUSION: This study provides comprehensive data on the prevalence and molecular epidemiology of the genetic diversity at the level of subtypes and alleles of Blastocystis sp. in different districts of Izmir province in Turkey. To the best of our knowledge, this is the first study which evaluates the distribution of subtypes and alleles of Blastocystis sp. according to PCR and SSU rRNA gene sequencing in patients with gastrointestinal complaints in different districts of Izmir province in Turkey.

The Risk Factors and Clinical Features of Acanthamoeba Keratitis: First Time Detection of Acanthamoeba T5 Genotype from Keratitis Patients in Turkey.

PURPOSE: The primary aim of this study is to investigate Acanthamoeba in clinical samples of keratitis cases (n = 60), in contact lens (CL) and lens care solutions of asymptomatic CL users (n = 41), and to identify the genotypes in positive samples. The secondary aim is to assess the risk factors and clinical features of Acanthamoeba keratitis (AK) patients. METHODS: All samples from patients and asymptomatic CL users were examined by microscopy and inoculated in non-nutrient agar plates. PCR was performed using the DNA isolated from corneal scrapings, CL and lens care solution samples. Positive DNA samples were sequenced to determine the genotype of Acanthamoeba. RESULTS: In none of the samples, Acanthamoeba was identified by microscopy, while Acanthamoeba was detected in a patient with keratitis by culture method. However, Acanthamoeba was detected in 11.66% (7/60) of the keratitis patients by PCR. The genotypes of these isolates detected by sequencing were T4 (4), and T5 (3). Acanthamoeba was detected in none of the samples of asymptomatic CL users by any of the three methods. CONCLUSION: To best of our knowledge, this is the first study to detect T5 genotype in AK patients from Turkey. In addition, the CL use was found to be an important risk factor for AK.

Genotyping and Molecular Identification of Acanthamoeba Genotype T4 and Naegleria fowleri from Cerebrospinal Fluid Samples of Patients in Turkey: Is it the Pathogens of Unknown Causes of Death?

PURPOSE: This study was aimed to investigate the presence of pathogenic free-living amoebae (FLA) in suspected cases of meningoencephalitis with unknown causes of death in Turkey. METHOD: A total of 92 patients, who were diagnosed as meningoencephalitis, were enrolled. All cerebrospinal fluid (CSF) samples were directly microscopically examined and cultured. Acanthamoeba, N. fowleri and B. mandrillaris were further investigated using molecular diagnostic tools including real-time PCR, sequencing, and phylogenetic analyses. RESULTS: The examined CSF samples were not found positive for the presence of FLA by microscopic examination and culture method. However, two CSF samples were detected positive by real-time PCR assay. Of the positive CSF samples, one was identified as Acanthamoeba genotype T4 and the second positive sample was identified as N. fowleri belonging to genotype II. Furthermore, the pathogens diagnoses was verified through Sanger sequencing. CONCLUSION: This study was significant to report the presence of Acanthamoeba genotype T4 and N. fowleri genotype II in CSF samples by real-time PCR assay. The present study shows the significance of primary amoebic meningoencephalitis (PAM) and granulomatous amoebic encephalitis (GAE) as one of the differential diagnoses to be considered by clinicians during the evaluation of suspected meningoencephalitis or cases of unknown cause in Turkey. Using real-time PCR, this has made the rapid detection, in a short time-frame, of Acanthamoeba and N. fowleri in CSF samples from patients. The problems with qPCR is that it is not available in every laboratory, reagents are expensive, and it requires skilled and expert personnel to set up these assays.

External Ophthalmomiasis Cases in Sirnak

Dermatological cases caused by myiasis make up approximately 20% of travel diseases. Ocular involvement occurred in approximately 5% of these myiasis cases. The conjunctival involvement of Oestrus ovis (O. ovis) larvae, which is the active agent in most ocular infestation cases in humans, causes external ophthalmomyiasis. External ophthalmomyiasis was diagnosed because of O. ovis first-stage larvae that were removed from the conjunctiva of three patients who applied to the Sirnak State Hospital Eye Clinic with complaints of foreign body sensation, burning, stinging, watering, and redness in the eyes and swelling of the eyelids in July and October. All symptoms of the patients disappeared within a few days after the removal of the larvae. Ophthalmomyiasis due to O. ovis must be kept in mind in the differential diagnosis of patients presenting with similar eye complaints in areas where sheep and goat husbandry is performed intensely.

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.

Investigation of Dientamoeba fragilis Prevalence and Evaluation of Sociodemographic and Clinical Features in Patients with Gastrointestinal Symptoms.

BACKGROUND: Dientamoeba fragilis is a protozoan parasite of the human gastrointestinal tract and still controversial in association with gastrointestinal symptoms. PURPOSE: We present cross-sectional study of the prevalence of D. fragilis, and sociodemographic and clinical features in the patients with gastrointestinal symptoms. METHODS: A total of 490 fecal specimens were collected from outpatients with gastrointestinal symptoms in the Department of Parasitology, Faculty of Medicine, Ege University and Celal Bayar University, Turkey. Fecal specimens were examined with microscopy and inoculated in Robinson medium. D. fragilis-positive samples were examined for the presence of other intestinal parasites using enzyme immunoassay. Real-time PCR analysis was performed on all samples. RESULTS: Of the 490 stool specimens examined by real-time PCR, 59 patients were positive for D. fragilis infection with prevalence rate of 12.04%. Forty-four of positive patients (74.5%) were found to be infected with only D. fragilis, while 23.7% were co-infected with Blastocystis and 1.7% were co-infected with Rotavirus. No statistically significant difference was found in all the examined patients in terms of D. fragilis positivity for all sociodemographic parameters. Loose stool consistency was associated with the presence of D. fragilis, with 18.3% (P = 0.001). When the clinical symptoms of all the patients participating in this study were examined, diarrhea was statistically more significant in patients with the presence of D. fragilis (16.3%; P = 0.001). The rate of diarrhea in D. fragilis-positive patients (84.09%; P = 0.0005) was higher than that of D. fragilis-negative patients and it was statistically significant. CONCLUSION: This study is important for assessing the prevalence of D. fragilis and its association with other factors in symptomatic patients in a large sample group in Turkey, as well as investigating the relationship of identified symptoms with the D. fragilis pathogenicity. It is suggested that D. fragilis in this case is not a commensal parasite but a pathogenic parasite and that the most common clinical symptom is diarrhea.

[Transverse myelitis associated with toxocariasis and the importance of locally produced antibodies for diagnosis]. / Toksokariyaza bagli bir transvers miyelit olgusu ve tanida yerel antikor üretiminin önemi.

Toxocariasis caused by Toxocara canis or less frequently by T.catis is a common parasitic infection worldwide. Clinical spectrum in humans can vary from asymptomatic infection to serious organ disfunction depending on the load of parasite, migration target of the larva and the inflammatory response of the host. Transverse myelitis (TM) due to toxocariasis is an uncommon illness identified mainly as case reports in literature. In this report, a case of TM who was diagnosed as neurotoxocariasis by serological findings has been presented. A 44-year-old male patient complained with backache was diagnosed as TM in a medical center in which he has admitted two years ago, and treated with pregabalin and nonsteroidal drugs for six months. Because of the progression of the lesions he readmitted to another center and treated with high dose steroid therapy for three months. After six months of follow up, improvement has been achieved, however, since his symptoms reccurred in the following year he was admitted to our hospital. Magnetic resonance imaging (MRI) examination revealed a TM in a lower segment of spinal cord. He was suffering with weakness and numbness in the left lower extremity. There was no history of rural life or contact with cats or dogs in his anamnesis. Physical examination revealed normal cranial nerve functions, sensory and motor functions. There has been no pathological reflexes, and deep tendon reflexes were also normal. Laboratory findings yielded normal hemogram and biochemical tests, negative PPD and parasitological examination of stool were negative for cysts and ova. Viral hepatitis markers, anti-HIV, toxoplasma-IgM, CMV-IgM, rubella-IgM, EBV-VCA-IgM, VDRL, Brucella tube agglutination, echinococcus antibody, autoantibody tests and neuromyelitis optica test were negative. Examination of CSF showed 20 cells/mm3 (mononuclear cells), 45 mg/dl protein and normal levels of glucose and chlorine. In both serum and CSF samples of the patient Toxocara-IgG antibodies were detected by Western blot (WB) assay. Low molecular weight bands (30-40 kDa) were detected in both of the samples by repeated WB testing. CSF revealed more intense bands suggesting local antibody production. Therefore the patient was diagnosed as neurotoxocariasis, and treated with steroid and mebendazole for six weeks. Clinical improvement was detected in the case and thoracic MRI revealed significant improvement in myelitis signs two months after treatment. In conclusion, toxocariasis should be considered in the differential diagnosis of TM although the involvement of central nervous system is rare and serological testing should be performed properly in the serum and CSF samples for the diagnosis.

[The molecular diagnosis of cryptosporidiosis in fresh and formalin preserved fecal samples]. / Taze ve Formaldehitle Saklanmis Diski Orneklerinde Cryptosporidiosisin Moleküler Tanisi.

The acid-fast staining method is widely used in the diagnosis of cryptosporidiosis, a disease causing diarrhea in humans. However in this technique, some of Cryptosporidium spp oocysts can not be stained and seen as formed "ghost-like bodies" and which can only be evaluated by experienced microscopists. In the recent years, PCR technique which is proven to be also highly sensitive in diagnosis and genotyping, is used as an alternative method. In our study we aimed to evaluate the efficacy of PCR in diagnosis of cryptosporodiosis. Thirty-three stool samples, belonging to 22 patients who applied to Ege University Hospital, Parasitology Clinic between August 2001 - August 2003 and microscopically diagnosed as cryptosporodiosis has been included in the study. Twenty-three of these 33 samples were processed immediately, while ten samples were stored in 10% formalin. As the control group, 11 stool samples including 8 specimens with different parasites and 3 negative samples were selected. Nested-PCR is applied to all samples. The sensitivity and specifity of the method for fresh and formalin preserved samples were found to be 100% and 50%, respectively. As a conclusion, PCR technique is found to be useful for diagnosis in cryptosporodiosis patients with especially those including few oocysts in their fresh feces.