The Importance of Antioxidant Enzymes and Oxidative Stress in Human Fascioliasis

Objective: The aim of this study was to determine the levels of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), and malondialdehyde (MDA) in the patients infected with Fasciola hepatica and establish whether these parameters differ among the patients with fascioliasis. Methods: The patient group consisted of 140 individuals with F. hepatica seropositive; the control group consisted of 140 healthy individuals who tested negative for this parasite and had no other diseases. The patient group consisted of individuals with no chronic diseases other than fascioliasis; in both the patient and the control groups, the subjects had no unhealthy habits such as smoking and alcohol consumption, etc. The blood samples taken to diagnose fascioliasis were evaluated by the ELISA method. The samples were studied according to the kit procedures for SOD, CAT, GPx and MDA markers. Results: In this study, 43.6% of 140 individuals in the patient group infected with F. hepatica had CAT (p=0.001), 35% had GPx (p=0.001), 12.9% had SOD (p=0.002), 90.7% had MDA (p=0.001). There was found a statistically significant difference between the patient and the control group in terms of the positivity of these four parameters. Conclusion: As a result, a statistically significant relationship was found between the increase in the SOD, GPx, CAT, and MDA levels and fascioliasis. The high rate of MDA revealed that oxidative stress occurred in patients with fascioliasis, resulting in an increased activity of SOD, GPx, and CAT. Amaç: Bu çalismanin amaci, Fasciola hepatica ile enfekte hastalarda süperoksit dismutaz (SOD), glutatyon peroksidaz (GPx), katalaz (CAT) ve malondialdehit (MDA) düzeylerini belirlemek ve fascioliasisli hastalarda bu parametrelerde farklilik meydana gelip gelmedigini ortaya koymaktir. Yöntemler: Hasta grubu, F. hepatica pozitif olan 140 hastadan; kontrol grubu ise bu parazit yönünden negatif bulunan ve baska herhangi bir hastaligi bulunmayan 140 saglikli kisiden olusturuldu. Hasta grubuna fascioliasis disinda herhangi bir kronik hastaligi olmayan ve hem hasta hem de kontrol grubuna sigara, alkol kullanimi olmayan kisiler dahil edildi. Hastalarda fascioliasis pozitifligini belirlemek için alinan kan örnekleri ELISA yöntemi ile çalisildi. Serum SOD, CAT, GPx ve MDA düzeyleri ELISA yöntemi ile degerlendirildi. Bulgular: Bu çalismada F. hepatica ile enfekte hasta grubundaki 140 kisinin %43,6'sinda CAT (p=0,001), %35'inde GPx (p=0,001), %12,9'unda SOD (p=0,002) ve %90,7 MDA pozitifligi saptandi. Bu dört parametrenin pozitifligi açisindan hasta ve kontrol grubu arasinda istatistiksel olarak anlamli fark bulundu (p=0,001). Sonuç: Sonuç olarak SOD, GPx, CAT ve MDA düzeyindeki artis ile fascioliasis arasinda istatistik olarak anlamli bir iliski saptanmistir. Çalismamizda MDA'nin yüksek oranda saptanmis olmasi fascioliasisli hastalarda oksidatif stres olustugunu ve SOD, GPx ve CAT aktivitelerinde artis oldugunu ortaya koymustur.

Detection of Blastocystis sp. in ulcerative colitis, Crohn's and chronic diarrheal patients by microscopy, culture and real-time polymerase chain reaction.

Blastocystis sp. is a zoonotic parasite that is common in humans and animals and settles in the large intestine. The parasite can cause various gastrointestinal system complaints such as indigestion, diarrhea, abdominal pain, bloating, nausea and vomiting. The aim of this study is to determine the distribution of Blastocystis in patients diagnosed with ulcerative colitis, Crohn's and diarrhea from the gastroenterology outpatient clinic and to compare the diagnostic value of the most preferred methods. A total of 100 patients, 47 men and 53 women, were included in the study. Of the cases, 61 had diarrhea, 35 had ulcerative colitis (UC) and four had Crohn's disease. Stool samples of the patients were analyzed by as direct microscopic examination (DM), culture and real-time polymerase chain reaction (qPCR). A total of 42% positivity was detected, 29% positivity in DM and trichrome staining, 28% in culture and 41% in qPCR. 40.4% (20/47) of men and 37.7% (22/53) of women were found to be infected. Blastocystis sp. was found in 75% of Crohn's, 42.6% of diarrheal and 37.1% of UC patients. Diarrhea cases are more common with UC and there is a close relationship between Crohn's and Blastocystis positivity. While DM and trichrome staining showed a sensitivity of 69%, PCR test was found to be the most sensitive diagnostic method with approximately 98%. Diarrhea and UC are often seen together. A close relationship was detected between Crohn's disease and Blastocystis. High prevalence of Blastocystis in cases with clinical symptoms reveals the importance of the parasite. There is a need for studies on the pathogenicity of Blastocystis sp. in various gastrointestinal cases, and it is thought that molecular techniques should be used since PCR is seen to be a much more sensitive.

IL-4, IL-10, TNF-α and IFN-γ Cytokine Levels in Patients with Cystic Echinococcosis and Fascioliasis.

Background: Cytokines are protein substances involved in immune response and inflammation regulation, cell growth, tissue repair and natural and adaptive immunity events such as hematopoiesis. Changes in Th1 and Th2 immune responses provide information on the clinical presentation, pathology and diagnosis of diseases. Methods: This study was conducted on 142 patients, aged 18-95 yr and applied to Parasitology Laboratory of Van Yuzuncu Yil Hospital between Aug 2018 and May 2019. Fifty-one of the patients were serologically diagnosed with CE, 53 with a diagnosis of fascioliasis, and 38 as the control group. Serum samples were taken from patients and levels of IL-4, IL-10, TNF α and IFN γ parameters were investigated by ELISA method. Results were evaluated by spectrophotometer and observed which immunologic parameters increased in which infections. Cytokine results were also evaluated according to patient age groups and genders. Chi-square and/or Fisher's exact test were used to evaluate the results. IL-4 response was detected in 50.9%; IL-10 in 44.2%; TNF-α in 43.3% and IFN-γ in 43.3% of CE seropositive patients. Results: IL-4, IL-10, TNF-α and IFN-responses detected in 50.9%, 44.2%, 43.3% and 43.3% of CE patients, and in 43%, 39.2%, 34.4% and 40.6% of fascioliasis patients, respectively. The presence of Th1 and Th2 responses were detected in both infections. There was a significant relationship between fascioliasis and IL-4 response and between CE and IL-4, IL-10 and TNF-α responses (P≤.05). Conclusion: These cytokine levels may provide information about the immune response to infections, and may be useful for early detection of disease and recurrence and monitoring of treatment.

Prevalência de la criptosporidiosis en seres humanos y terneros, y detección molecular del Cryptosporidium parvum / Prevalence of cryptosporidiosis in humans and calves and molecular detection of Cryptosporidium parvum

RESUMEN Objetivo. El objetivo de este estudio fue investigar la prevalencia de especies de Cryptosporidium en humanos y terneros en la provincia de Van, Turquía. Materiales y métodos. Se incluyeron en el estudio un total de 150 pacientes, incluidos 50 pacientes en hemodiálisis, 40 pacientes inmunosuprimidos con diarrea, 30 pacientes con diarrea solamente y 30 pacientes inmunocompetentes. Se recolectaron muestras de heces rectales de un total de 50 terneros alojados en establos y granjas en 10 aldeas centrales de Van, Turquía. Resultados. Se detectó Cryptosporidium parvum en el 17.3% de las 150 muestras de heces tomadas de seres humanos. C. parvum se observó en el 20% de los 50 pacientes en hemodiálisis, el 32.5% de los 40 pacientes inmunosuprimidos con diarrea y el 10% de los 30 pacientes con diarrea solamente, mientras que no hubo Cryptosporidium spp. detectado en los pacientes inmunocompetentes. C. parvum se observó en sólo el 6% de los 30 terneros diarreicos. Conclusiones. Claramente se entendio que la Criptosporidiosis fue detectada en una alta tasa en las muestras de los pacientes inmunosuprimidos sin y con sintomas de diarrea, y que además la especie activa que causó la enfermedad fue el agente etiologico Criptosporidium parvum. Por lo tanto, estos dos grupos de pacientes deben ser evaluados en lo que a términos de Criptosporidiosis se refiere.

ABSTRACT Objective. To investigate of the prevalence of Cryptosporidium species in humans and calves in the province of Van, Turkey. Materials and methods. Included in this research were 150 patients, comprising 50 hemodialysis patients, 40 immunosuppressed patients with diarrhea, 30 patients with diarrhea only, and 30 immunocompetent patients. Collected were stool rectal samples from 50 calves that were housed in stables and farms in 10 central villages of Van, Turkey. Results. Cryptosporidium parvum was detected in 17.3% of the 150 human stool samples. C. parvum was observed in 20% of the 50 samples from the hemodialysis patients, 32.5% of the 40 samples from the immunosuppressed patients with diarrhea, and 10% of the 30 samples from patients with diarrhea only, whereas no Cryptosporidium spp. was detected in the samples from the immunocompetent patients. C. parvum was observed in only 6% of the samples from the diarrheic 30 calves. Conclusions. It was clearly understood that cryptosporidiosis was detected at a high rate in the samples from the immunosuppressed patients and those who were immunosuppressed with diarrhea, and that the active and effective species that causes cryptosporidiosis in the Van region is C. parvum. Hence, these patient groups should be evaluated in terms of cryptosporidiosis.

Evaluation of Formalin-fixed Paraffin-embedded Tissue Samples Diagnosed by Histopathology as Echinococcus in Çorum

Objective: This study aimed to detect the presence of Echinococcus spp. in formalin-fixed paraffin-embedded (FFPG) samples of hydatid cyst cases and to discuss the DNA isolation problems in FFPG samples. Methods: FFPG samples of 47 cases diagnosed with hydatid cyst were included in this study. Demographic characteristics of the cases were investigated. Microtome sections were taken from the samples and deparaffinization, DNA extraction, polymerase chain reaction (PCR), and gel agarose electrophoresis procedures were performed. Results: Of the cases, 55.3% were female, whereas 45.7% were male. Average age was 45.47 and 68.1% of the cases were located in the liver, 17.0% in the lung, 12.8% in the abdomen, and 2.1% in the brain. DNA was obtained in only 11 (23.4%) of the FFPG cyst samples and no proliferation was detected in the PCR products of any of the sample. Conclusion: The scolex/germinal membrane' absence in the FFPG sections, intense inflammatory cell reaction, presence of fibrosis and stromal/parenchymal tissue, DNA damage due to formaldehyde action, long-term archiving, and insufficient amount of DNA obtained were considered as factors preventing DNA replication in PCR.

Molecular characterization of human lung and liver cystic echinococcosis isolates in Van Province, Turkey.

Cystic echinococcosis (CE) is a zoonotic infection and could lead to significant public health problems. The genetic diversity of CE includes five species: E. granulosus sensu stricto (s.s.) (G1-G3), Echinococcus equinus (G4), Echinococcus ortleppi (G5), Echinococcus canadensis genotypic cluster (G6, G7, G8 and G10, with the doubtful G9) and the Echinococcus felidis (lion strain). The species are important in epidemiology, pathology, control, prevention measures and vaccine/drug designs. The aim of the present study was to determine the E. granulosus genotypes in humans in the Van province in east of Turkey. In total, 102 echinococcal cysts were collected from operated patients. Genomic analyses were conducted with PCR-RFLP of the rDNA internal transcribed spacer 1 (ITS1) fragment and partial PCR sequencing of the cytochrome c oxidase subunit 1 (cox1) mitochondrial DNA gene region. In total, DNAs of 96 isolates could be extracted, unfortunately six extractions failed. The PCR-RFLP analysis findings were identical in all isolates. Two bands were observed at approximately 300 bp and 600 bp. All profiles corresponded to the G1-G3 strain. Also, 446 bp amplified gene regions were observed for cox1. Out of 20 samples, alignment of 16 sequences exhibited a total identification (100%) of granulosus sensu stricto (G1/G3). Of 16 samples, 8 were obtained in the lung and 12 were obtained in the liver; 8 belonged to male and 12 belonged to female patients. Other four samples exhibited one nucleotide substitution at different positions. Four samples had one nucleotide substitution at different positions. We detected single nucleotide variations in TRH1, TRH67, TRH85 and TRH89 isolates at the positions C240T; G330T; G211A and T157C, respectively. In conclusion, the present study was the first comprehensive molecular investigation on genetic characterization of human CE isolates in Van region. The findings demonstrated that E. granulosus s.s. was the dominant species, which indicated that the sheep-dog cycle was the source in human infections. And, probably, it would be possible to describe these mutations as "Turkey" or "lung" variants. In addition to contributing molecular epidemiological data, the present results should be considered when designing and implementing E. granulosus control programs.

A Rare Case of Secondary Hydatid Cyst: Uterus and Colon Locations in the Same Patient

The aim of the present study was to present a case with secondary hydatid cysts in both uterus and colon. The patient was a 71-year-old female living in Hakkari, Turkey. She was admitted to the Van Yuzuncu Yil University Faculty of Medicine Medical Center with complaints of chronic abdominal and pelvic pain, and swelling in the abdomen. First, the sagittal T2 weighted magnetic rezonance imaging (MR) showed a type-3 cyst hydatid with daughter vesicles located at the posterior of uterus. Later, MR revealed a type-2 cystic lesion with detached membrane adhered to the anterior wall of colon and it was reported to be associated with abdomen. When the previous liver surgery history of the patient was kept in mind, the new finding was suggestive of a secondary cystic hydatid . In conclusion, it is possible to diagnose secondary cystic echinococcosis in patients with a history of primary cyst surgery in liver or any other organ by combining the symptoms and imaging findings.

A Comprehensive Retrospective Study: Intestinal Parasites in Human in Van Province

Objective: The aim of present study was to retrospectively determine the prevalence of intestinal parasites in patients, who were referred to Parasitology Laboratory in Van Yüzüncü Yil University, Faculty of Medicine during an 11-year period. Methods: Stool samples of 69633 individuals admitted to the outpatient clinics in the hospital were initially evaluated with native-Lugol, and then by flotation, sedimentation, trichrome staining, modified acid-fast staining and ELISA methods. Results: Twenty-four parasite species were identified in parasite-positive patients. At least one or more parasite species were found in 34.1% of all patients. The most commonly observed parasite was Blastocystis hominis (26.5%). Among pathogen parasites, Giardia intestinalis (G. intestinalis) was detected in 9.3%, Ascaris lumbricoides (A. lumbricoides) was detected in 2.5%, Entamoeba histolytica/Entamoeba dispar in 0.8%, Cystoisospora belli in 0.004%, Fasciola hepatica in 0.04%, Dicrocoelium dendriticum in 0.001%, Strongyloides stercoralis in 0.001% and hookworm in 0.001% of the patients. Conclusion: It was determined that, pathogen parasites such as G. intestinalis and A. lumbricoides were still observed at high rates in Van province, especially in children, and the problem of parasitosis is still continuing, although the prevalence of parasites has declined when compared to the previous years.

Cystoisospora sp. Infection Determined in Immunosuppressed and Immunocompetent Children: Three Cases Report.

The aim of this study was to present three cystoisosporiasis cases diagnosed in pediatric patients of the Yuzuncu Yil University Medical Faculty. In the study, stool samples of the patients were evaluated by native-Lugol and modified acid-fast staining methods in the Parasitology Laboratory. The first case was a 4-year-old male child diagnosed with acute lymphoblastic leukemia (ALL). It was reported that the patient had abdominal pain, and permanent bloody and mucous diarrhea (8-10 times a day) was present for almost 1 week after the beginning of ALL treatment. The second case was a 10-year-old boy diagnosed with depression. The patient was brought to our hospital by his parents with complaints of abdominal pain, diarrhea, lack of appetite, weight loss, and fatigue persisting since 1 month in addition to headache, fear, sleeplessness, and waking up with cry. The third case was a 13-year-old boy who complained of abdominal pain, diarrhea (rare occasions), lack of appetite, and headache for 2 months. These patients had not traveled abroad. The cases were treated successfully with co-trimoxazole. Our results suggest that all patient groups with diarrhea and abdominal pain should also be considered in cystoisosporiasis.

In Vitro Efficacy of the Ankaferd Galenic Hemostatic Extract as a Germicidal Agent.

BACKGROUND: Hydatid cysts are encountered frequently in regions endemic with livestock. The basic treatment for a hydatid cyst is total surgical removal of the cyst and its inner contents. Hypertonic NaCl or diluted betadine solution are used as germicidal agents for most hydatid surgeries. However, the germicidal efficacy of the Ankaferd Blood Stopper® (ABS) has not been investigated. Thus, we compared the efficacy of ABS for hydatid cysts with that of other germicidal agents. METHODS: Lung and liver tissues containing hydatid cyst liquid were collected from slaughterhouses. Six samples of each cyst were randomly allocated into different groups as follows: 20% hypertonic NaCl, betadine solution, ABS, 20% liquefied Andazole solution, 0.1% eosin, and distilled water. All groups were examined microscopically at 5, 10, and 15 min after treatment began to determine protoscolece viability rates. RESULTS: The most efficacious germicidal agent at 5 min was ABS, and betadine and hypertonic NaCl had similar efficacies. Betadine, ABS, and hypertonic NaCl showed similar efficacies at 15 min. CONCLUSION: ABS was an effective germicidal agent to treat hydatid cysts.

[Cystic Echinococcosis Seropositivity in the Blood Samples Sent to Parasitology Laboratory of Yüzüncü Yil University Medical Faculty between 2005 and 2013: Retrospective Assessment]. / Yüzüncü Yil Üniversitesi Tip Fakültesi Parazitoloji Laboratuvarina 2005-2013 Yillari Arasinda Gönderilen Kan Örneklerinde Kistik Ekinokokkozis Seropozitifligi: Retrospektif Degerlendirme.

OBJECTIVE: This study was performed in order to contribute the data on the prevalence of cystic echinococcosis (CE) in Turkey and to reveal this parasitosis problem in Van province. METHODS: Blood samples of 2642 patients (1214 men, 1428 women; 506 of them 0-13 age, 2136 of them 14 and over age group), which were sent with suspected CE to Parasitology Laboratory from various policlinics of Yüzüncü Yil University Medical Faculty, between the dates of 01.09.2005 and 01.09.2013, were evaluated for CE by using Enzyme-Linked Immunosorbent Assay (ELISA) (R-Biopharm, Germany; IgG) method. RESULTS: 801 (30.3%) of examined blood samples of these patients were found positive for this parasitosis. Seropositivity was found 31.9% of men, 29% of women; 33.4% of 0-13 age group, 29.6% of 14 and over age group in the study. Cysts, which were obtained from operated seropositive patients, were confirmed that unilocular cysts as pathological. CONCLUSION: As a result, it was seen that CE is continued to be a major public health problem in Van province and it was appear that comprehensive protection and control programs should be carry out to struggle this disease.

[Evaluation of Cystic Echinococcosis Suspected Patients Applied to National Parasitology Reference Laboratory of Public Health Institution of Turkey Between 2009-2013]. / Gaziantep Üniversitesi Tip Fakültesi Leishmaniasis Tani ve Tedavi Merkezine Basvuran Kutanöz Leishmaniasis Olgularinin Degerlendirilmesi.

OBJECTIVE: Cystic echinococcosis (CE) caused by the metacestode form of Echinococcus granulosus is an important public health problem common in our country. In this study, anti-E. granulosus antibodies were aimed to investigate in the serum samples of CE suspected patients who applied to the National Parasitology Reference Laboratory of Public Health Institution of Turkey. METHODS: In the study, serum samples of 2921 patients which were sent to our laboratories from different hospitals between 1 January 2009 and 31 December 2013 were evaluated with at least one of the following tests: Enzyme-Linked Immunosorbent Assay (ELISA), Indirect Hemaglutination Assay (IHA) and Western Blot (WB) techniques. RESULTS: Four hundred thirty nine (15.03%) of inspected 2921 samples were determined seropositive with at least one of the methods. When the results analyzed by gender, 13% of males and 16.40% of females were found positive. Examined the distribution of the results by years, with a maximum of 25% positivity was observed in 2009. Compatibility was determined at the rate of 91.4% among ELISA and IHA results; also 89.7% among WB and the other tests results. CONCLUSION: Despite the gradual decreases the CE in Ankara and its surroundings, it is still continues to be a major public health problem. Essential prevention and control measures should be taken to reduce the prevalence of the disease. Also in the diagnosis of the disease, more reliable results can obtained with applying two tests (ELISA/IHA) together and confirm the positivity with WB.

[Giardia muris infection in laboratory rats (Rattus norvegicus) and treatment with metronidazole]. / Laboratuvar Siçanlarinda (Rattus norvegicus) Giardia muris Enfeksiyonu ve Metronidazol ile Sagaltimi.

OBJECTIVE: This study was conducted to determine the effectiveness of metronidazole for treatment of Giardia muris infection in laboratory rats. METHODS: The feces of rats was yellow watery diarrhea and brought to the surgery research center of University of Ondokuz Mayis in order to be a study. Stool samples were examined by native examination, evaluation of infection rates was done with an X40 lens, and results were recorded as positive from 1 to 4. Metronidazole was administered to infected animals orally for 5 days with a 20 mg/kg dose. RESULTS: As a result of fecal examination of 64 rats held in groups of four in cages, 15 of the cages (60 rats) were found to be infected with G. muris. While agents were not observed in collected stool samples following 5, 7, and 14 days of drug administration of 14 groups, trophozoite density in one cage was decreased (75%), and adverse effects were not seen in rats. CONCLUSION: Metronidazole was found to be an effective drug for the treatment of giardiasis.

Helminths of roe deer (Capreolus capreolus) in the Middle Black Sea Region of Turkey.

Fifteen roe deer were examined at necropsy from Northern Turkey in the period 2006-2010 for the helminth infections. Totally 6470 helminth specimens were collected and identified by morphological criteria. Twenty-five helminth species were identified (1 of the Class Trematoda, 1 of Cestoda and 23 of Nematoda). Dicrocoelium dendriticum (Prevalence 20%) was found in liver. Cysticercus tenuicollis (6.6%) was found in mesentery. Haemonchus contortus (53.3%), Ostertagia leptospicularis (73.3%), O. leptospicularis (minor morph: kolchida) (53.3%), Ostertagia ostertagi (26.6%), Spiculopteragia spiculoptera (66.6%), S. spiculoptera (minor morph: mathevossiani) (6.6%), Teladorsagia circumcincta (40.0%), T. circumcincta (minor morph: davtiani) (6.6%), T. circumcincta (minor morph: trifurcata) (6.6%), Trichostrongylus axei (66.6%) were found in abomasum. Trichostrongylus andreevi (6.6%), T. colubriformis (6.6%), T. longispicularis (26.6%), T. vitrinus (40.0%), T. capricola (6.6%), Cooperia oncophora (26.6%), C. punctata (6.6%), Nematodirus filicollis (66.6%), and Capillaria bovis (26.6%) were found in small intestine. Oesophagostomum venulosum (46.6%), Chabertia ovina (26.6%), and Trichuris ovis (13.3%) were found in large intestine. Dictyocaulus capreolus (6.6%) was found in lungs.

Molecular characterization and prevalence of cystic echinococcosis in slaughtered water buffaloes in Turkey.

The present study was carried out between March 2006 and June 2010. During the study nine abattoirs were visited and 166 water buffalo internal organs were examined in Black Sea Region of Turkey. It was found that 10.24% buffaloes were infected with cystic echinococcosis (CE). The rate of CE found as 3.77% in males and 21.66% in females, 37.93% in older and 4.38% in young animals. The degree of prevalence according to age and sex was statistically significant (p<0.05). CE was observed 29.41% only in liver, 47.06% only in lungs and 23.53% in both liver and lungs. Therefore, the lungs were the predominant sites of the CE in buffaloes. Molecular identification on nine isolates, based on mitochondrial cox1 sequencing analyses, revealed that six cysts belonged to G1 genotype (domestic sheep strain) while 3 samples showed variant genotypes of Echinococcus granulosus complex G1-G2-G3. Two of them showed a thymine in position 52, like G2 strain, but the rest of sequences were completely identical to strain G1; also one specimen showed a single nucleotide change compared to strain G1 (C122T).