Vermamoeba vermiformis (V. vermiformis) is one of the most common free-living amoeba (FLA) and is frequently found in environments such as natural freshwater areas, surface waters, soil, and biofilms. V. vermiformis has been reported as a pathogen with pathogenic potential for humans and animals. The aim is to report a case of non-Acanthamoeba keratitis in which V. vermiformis was the etiological agent, identified by culture and molecular techniques. Our case was a 48-year-old male patient with a history of trauma to his eye 10 days ago. The patient complained of eye redness and purulent discharge. A slit-lamp examination of the eye revealed a central corneal ulcer with peripheral infiltration extending into the deep stroma. The corneal scraping sample taken from the patient was cultured on a non-nutritious agar plate (NNA). Amoebae were evaluated according to morphological evaluation criteria. It was investigated by PCR method and confirmed by DNA sequence analysis. Although no bacterial or fungal growth was detected in the routine microbiological evaluation of the corneal scraping sample that was cultured, amoeba growth was detected positively in the NNA culture. Meanwhile, Acanthamoeba was detected negative by real-time PCR. However, V. vermiformis was detected positive with the specific PCR assay. It was confirmed by DNA sequence analysis to be considered an etiological pathogenic agent. Thus, topical administration of chlorhexidine gluconate

OBJECTIVEAcanthamoeba, one of the free-living amoeba, has been detected in many environmental samples, mainly in water, soil and air. Acanthamoeba keratitis and granulomatous amoebic encephalitis are among the most important clinical manifestations caused by Acanthamoeba. In this study, it was aimed to determine the sensitivity of the rapid loop mediated isothermal amplification (LAMP) test designed with primers specific to Acanthamoeba 18S rRNA gene to detect more rapidly the presence of Acanthamoeba in clinical and environmental samples.METHODSAcanthamoeba strain grown in culture was diluted in 200 μL as 1x106 trophozoites and DNA was isolated, and the amount of DNA was determined by Nano-Drop Spectrophotometer. The purified DNAs were diluted from 1000 pg to 0.001 pg and used in colorimetric and fluorescence-based LAMP reactions. The LAMP reaction mixture was incubated for 60 minutes at 63 °C in a total volume of 25 μL.RESULTSTo determine the sensitivity of the test, positivity of Acanthamoeba genomic DNA was observed at 1, 10, 100 and 1000 pg/reaction in both colorimetric and fluorescence-based LAMP tests. The lowest analytical sensitivity of both calorimetric and fluorescent LAMP assay was determined as 1 pg/reaction. In addition, as a result of LAMP reaction applied with other parasite DNAs to evaluate the specificity of the test, no LAMP product was detected in calorimetric and 1% agarose gel electrophoresis, except for positive control, and the specificity of the test was determined as 100%.CONCLUSIONIt has been demonstrated that the LAMP assay designed by targeting 18S rRNA gene of Acanthamoeba has a detection limit of 1 pg of genomic DNA. It is promising that LAMP test is more sensitive and faster than culture method, as well as simple, inexpensive and highly sensitive. For this reason, it is thought that developed test can be applied in the diagnosis of Acanthamoeba in environmental and clinical samples.

Acanthamoeba spp., Balamuthia mandrillaris, and Naegleria fowleri are pathogenic free-living amoeba (FLA) and are commonly found in the environment, particularly soil. This pathogenic FLA causes central nervous system -affecting granulomatous amebic encephalitis (GAE) or primary amebic meningoencephalitis (PAM) and can also cause keratitis and skin infections. In the present study, we aimed to determine the quantitative concentration of Acanthamoeba spp., B. mandrillaris, and N. fowleri in soil samples collected from places where human contact is high by using a qPCR assay in Izmir, Turkey. A total of 45.71% (n = 16) of Acanthamoeba spp., 20% (n = 7) of B. mandrillaris, and 17.4% (n = 6) of N. fowleri were detected in five different soil sources by the qPCR assay. The quantitative concentration of Acanthamoeba spp., B. mandrillaris, and N. fowleri in various soil sources was calculated at 10 x 105 -6 x 102, 47 x 104 to 39 x 103, and 9 x 103 - 8 x 102 plasmid copies/gr, respectively. While the highest quantitative concentration of Acanthamoeba spp. and B. mandrillaris was determined in garden soil samples, N. fowleri was detected in potting soil samples. Three different genotypes T2 (18.75%), T4 (56.25%), and T5 (25%) were identified from Acanthamoeba-positive soil samples. Acanthamoeba T4 genotype was the most frequently detected genotype from soil samples and is also the most common genotype to cause infection in humans and animals. To the best of our knowledge, the present study is the first study to identify genotype T5 in soil samples from Turkey. In conclusion, people and especially children should be aware of the hidden danger in the garden and potting soil samples that come into contact most frequently. Public health awareness should be raised about human infections that may be encountered due to contact with the soil. Public health specialists should raise awareness about this hidden danger in soil.

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 İzmir, 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 İzmir, 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 İzmir. 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 İzmir 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 İzmir province 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.

Naegleria fowleri, the causative agent of primary amoebic meningoencephalitis (PAM), is a free-living amoeba. It is a water-borne infection usually detected in children and young people with healthy immune system who swim, dive and perform activities in fresh and hot springs. In this study, it was aimed to raise awareness in the differential diagnosis of meningitis etiopathogenesis by showing that N. fowleri may also be the causative agent, albeit very rarely, in meningitis cases in Turkey. Our case was an 18-year-old male patient whose relatives stated that he has gone to the hot spring; his headache complaint started after 2 to 3 days after return from the hot spring. Cerebrospinal fluid (CSF) sample taken from the patient was investigated by direct microscopic examination, real-time PCR method and sequence analysis. The CSF sample collected was taken into distilled water considering the possibility of transformation of trophozoites to intermediate form and incubated at 37 °C for 1 to 2 h, and pear-shaped non-permanent flagellated forms were observed in the direct microscopic examination, and molecular typing was performed to confirm the diagnosis. This study was a comprehensive case of N. fowleri whose etiological agent was isolated and confirmed by real-time PCR in Turkey. Clinician awareness would be the key factor in correctly diagnosing PAM. It is also recommended to investigate all likely environmental water sources in Turkey for more detailed information on the distribution and molecular identification of Naegleria species, ultimately to evaluate the potential pathogenic threat to human health and to develop strategies to combat such threats.

Ehrlichia spp. is tick-borne zoonotic pathogen that can infect humans and animals. Nowadays, among the tests used in the diagnosis of ehrlichiosis, the importance of molecular methods is increasing steadily due to their high sensitivity and specificity. The aim of this study was to determine the analytical sensitivity of a conventional polymerase chain reaction (PCR) targeting Ehrlichia spp. disulfide oxidoreductase (DSB) gene. Ehrlichia spp. DSB gene was cloned into the TOPO vector. After TOPO plasmid containing DSB gene were serially diluted, PCR targeting the Ehrlichia spp. DSB gene was performed. While working on this research, blood and skin scraping samples of a stray dog clinically suspected with leishmaniasis as well as treated for leishmaniasis arrived to our laboratory. Thereafter, PCRs targeting Ehrlichia spp. DSB and 16S rRNA and Leishmania kinetoplast DNA (kDNA) genes were performed to identify the pathogen in blood and skin scraping samples of the stray dog. The analytical sensitivity of the PCR assay targeting Ehrlichia spp. DSB gene was 1 ≥ copy plasmid/reaction using serially diluted TOPO plasmid containing DSB gene. PCR targeting the Ehrlichia spp. DSB gene was positive and PCR targeting Leishmania spp. kDNA was negative in blood and skin samples of the stray dog clinically suspected with leishmaniasis. Using nested PCR targeting Ehrlichia spp. 16S rRNA, E. canis was identified in blood and skin scraping samples of the stray dog. In this study, PCR targeting Ehrlichia spp. DSB gene has been shown to have high sensitivity. Also it was shown molecular methods can help clinicians in differential diagnosis of ehrlichiosis and leishmaniasis to prevent inappropriate treatment.

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.

Blastocystis is a single-celled parasite commonly found in humans and its pathogenic role is still controversial. In recent years, some studies have suggested that Blastocystis may be a possible agent of gastrointestinal and dermatological symptoms such as acute or chronic urticaria, angioedema, rash, itch, palmoplantar, and diffuse pruritus. We aimed to investigate whether there is a relationship between Blastocystis subtypes and alleles in patients with chronic spontaneous urticaria (CSU) as a case-control study. In this study, stool samples were collected from patients with CSU (n=135) and healthy individuals (n=54). The presence of Blastocystis was investigated using the direct saline smear, Lugol's iodine staining, trichrome staining, Jones' medium culture and PCR assays in stool samples and subtypes (STs) were determined by sequencing according to DNA barcoding. The presence of Blastocystis was identified in 30.4% (64/210) the stool samples, including 31.9% (43/135) of the patients with CSU and 14.8% (8/54) of the control group. Moreover, it was found statistically significant the presence of Blastocystis in terms of both groups (p<0.018). ST3 was detected in 45.9% and 62.5 % as the most prevalent subtype the patients with CSU and the control group, respectively. ST1 (18.9%), ST2 (27%) and ST7 (8.1%) was identified in the patients with CSU group. There was no statistically significant correlation between Blastocystis subtypes and both the groups (p<0.240, p<0.323). Allele 4 for ST1; alleles 9, 10, 11 and 12 for ST2; alleles 34, 36 and 38 for ST3; alleles 41 and 101 for ST7 were detected. Allele 34 (ST3) was found significant in the patients with CSU as compared with control group (p<0.020). Moreover, statistically significant association was found between total IgE value and the certain subtypes (ST2 and ST3) (p<0.0001). As a result of this study, the presence of Blastocystis ST3 allele 34 significantly associated with chronic spontaneous urticaria was revealed.

Free-living amoeba (FLA) is widely distributed in the natural environment. Since these amoebae are widely found in various waters, they pose an important public health problem. The aim of this study was to detect the presence of Acanthamoeba, B. mandrillaris, and N. fowleri in various water resources by qPCR in Izmir, Turkey. A total of (n = 27) 18.24% Acanthamoeba and (n = 4) 2.7% N. fowleri positives were detected in six different water sources using qPCR with ITS regions (ITS1) specific primers. The resulting concentrations varied in various water samples for Acanthamoeba in the range of 3.2x105-1.4x102 plasmid copies/l and for N. fowleri in the range of 8x103-11x102 plasmid copies/l. The highest concentration of Acanthamoeba and N. fowleri was found in seawater and damp samples respectively. All 27 Acanthamoeba isolates were identified in genotype level based on the 18S rRNA gene as T4 (51.85%), T5 (22.22%), T2 (14.81%) and T15 (11.11%). The four positive N. fowleri isolate was confirmed by sequencing the ITS1, ITS2 and 5.8S rRNA regions using specific primers. Four N. fowleri isolates were genotyped (three isolate as type 2 and one isolate as type 5) and detected for the first time from water sources in Turkey. Acanthamoeba and N. fowleri genotypes found in many natural environments are straightly related to human populations to have pathogenic potentials that may pose a risk to human health. Public health professionals should raise awareness on this issue, and public awareness education should be provided by the assistance of civil authorities. To the best of our knowledge, this is the first study on the quantitative detection and distribution of Acanthamoeba and N. fowleri genotypes in various water sources in Turkey.

It is not clear that Blastocystis remains without damage to the digestive tract or has a pathogenic effect in relation to subtypes in immunocompromised people, such as cancer patients. The present study aimed to investigate the frequency and subtype distribution of Blastocystis in cancer patients who were followed-up and treated in the Oncology clinic of Firat University Hospital and to determine the clinical signs of infected sufferers. 201 patients aged ≥ 18 with a diagnosis of cancer were enrolled in this cross-sectional study. Patients’ stool samples were examined between September 2017 and August 2019 by native-Lugol, trichrome staining. Microscopy-positive stool samples were subjected to DNA isolation and subtyped by Sequence Tagged Site (STS)-PCR analysis. The symptoms and demographic characteristics of the patients were also evaluated. Totally, 29 (14.4%) samples were positive for Blastocystis after all methods. 15 (51.7%) out of 29 samples were successfully subtyped by the sequenced-tagged site(STS)-PCR, while 14 (48.3%) could not be typed. Three subtypes of Blastocystis were detected: ST3 (40%), ST2 (33%), ST1 (20%), and one mixed infections with ST1/ST2 (6%). There was no statistically significant difference in terms of clinical findings and demographic characteristics. The outcomes of our study promote the idea that Blastocystis could be an asymptomatic and harmless commensal organism. However, more comprehensive molecular and clinical studies are needed to fully determine the pathogenicity and epidemiology of Blastocystis in cancer patients.

Dirofilariasis is a vector-borne disease that is present worldwide. This report describes a giant subconjunctival granuloma which mimics scleritis, caused by D. immitis. A 60-year-old man was referred with the complaints of irritation, redness, and swelling at the medial part of the right eye. He was living in Izmir province located in western Turkey. Slit-lamp examination showed a firm, immobile mass measuring 13.0 × 5.0 × 5.0 mm with yellowish creamy color. The mass was completely removed surgically under local anesthesia mainly for diagnosis. Histopathology revealed typical morphological features of a filarioid nematode in favor of Dirofilaria as characterized by the external smooth cuticular surface, cuticular layer, muscle layer, and intestinal tubule. Molecular study was performed using DNA isolated from paraffin-embedded tissue sections of the worm. PCR amplification and then DNA sequence analysis of cytochrome c oxidase subunit 1 (cox1) gene fragment confirmed that the worm was D. immitis. It is suggested that this may represent the first human case of D. immitis occurring in subconjunctival granuloma in Turkey. Although rare, D. immitis caused by ocular dirofilariasis in humans should be considered.

Objective: Blastocystis is a common protozoan parasite detected worldwide. This study aims to investigate the in vitro inhibitory effect of ethyl alcohol extracts of Tunceli garlic (Allium tuncelianum) on growth of Blastocystis ST3 subtype. Methods: Blastocystis ST3 were grown in 3 ml Jones's medium containing A. tuncelianum prepared at five different concentrations for 72 hours at 37 degrees C. Distilled water was used as negative control whereas metronidazole (MTZ) and trimethoprim-sulfamethoxazole (TMP-SMX) prepared in 0.1, 0.5 and 1 mg/ml of concentrations was used as the positive control. Results: According to the results, the MIC90 of the A. tuncelianum was 115 mg/ml at 72 hours. MIC90 of MTZ was 1 mg/ml at 72 hours (94.96%). TMP-SMX had the highest growth MIC90 effect which was reached at 1 mg/ml concentration during 24 hours (88.18%). The growth inhibitory effect of A. tuncelianum extract increased gradually in all doses and timepoints. Conclusion: In this study, obtained findings show the potential inhibitory effect of A. tuncelianum plant extract against Blastocystis ST3 which causes symptomatic diarrhea in humans. It may be recommended to consume the whole A. tuncelianum plant as food supplement. A. tuncelianum extract could be considered as an alternative for the potential treatment of Blastocystis infection.

BackgroundDientamoeba fragilis is a protozoan parasite of the human gastrointestinal tract and still controversial in association with gastrointestinal symptoms.PurposeWe present cross-sectional study of the prevalence of D. fragilis, and sociodemographic and clinical features in the patients with gastrointestinal symptoms.MethodsA 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.ResultsOf 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.ConclusionThis 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.

Calprotectin is a protein that is mostly released from neutrophils, monocytes, macrophages and submucosal epithelial cells. Fecal calprotectin (f-CP) is a marker of intestinal inflammation. There are some discussions about the pathogenicity of D. fragilis in the gastrointestinal tract. In this study, we investigated whether f-CP level is a factor supporting the pathogenicity of D. fragilis. The f-CP levels were evaluated in patients with only D. fragilis positive in comparison with healthy controls. Moreover, the levels of f-CP were investigated in fecal samples of D. fragilis negative patients with gastrointestinal complaints. The fecal samples were collected from three groups. Three groups of fecal samples were examined directly microscopy, trichrome staining, cultivation, enzyme immunoassay (EIA) and real-time PCR assay. In the first group (Group 1, n = 34), patient stool samples with gastrointestinal symptoms (without other pathogens) found only with D. fragilis were included. In the second group (Group 2, n = 31), there were patients' stool samples with gastrointestinal symptoms that D. fragilis was negative (but there may be other pathogenic agents). In the control group (Group 3, n = 23), we used fecal samples collected from healthy volunteers without any infection or gastrointestinal complaints. The collected fecal samples were stored at -20 °C until analysis. Levels of f-CP were determined by using human calprotectin ELISA kits. Total of 88 patients were enrolled in three different groups. We obtained f-CP levels as follows: 33.40 ng/mg protein in the group 1, 15.99 ng/mg protein in the group 2 and 1.54 ng/mg protein in the group 3. Statistically significant difference in f-CP levels of the group 1 and the group 2 were obtained when compared with healthy controls (p < 0.0001). However, the f-CP levels of the group 1 were not significantly different from the group 2 (p > 0.99). In conclusion, increased levels of f-CP are shown as a marker of an inflammatory disease of the lower gastrointestinal tract in infected humans. There is continues controversy about the pathogenicity of D. fragilis in symptomatic and asymptomatic patients. The findings of this study contribute to the ongoing debate about the pathogenicity of D. fragilis. In our study, the potential pathogenicity of D. fragilis is associated with increased f-CP concentrations with parasite detection in the fecal samples and therefore we assume that the parasite is not only a harmless commensal. In summary, higher levels of f-CP found in D. fragilis positive patients suggest the importance of researches that support the pathogenicity of indicated parasite.

Over the past decade, the number of international travels has increased. Hence, the risk of transmission of parasitic diseases has also increased. One of the risk infections is malaria; Plasmodium vivax and P. falciparum species can be transmitted. The distribution of leishmaniasis cases has been reported from the south of USA to the north of Argentina. Approximately 57,000 cases of cutaneous and mucocutaneous leishmaniasis occur annually, and approximately 4000 visceral leishmaniasis cases are observed. It is reported that Chagas disease is endemic in 21 countries, and approximately 6 million people are affected every year. In this continent, 25 million people are at a risk of schistosomiasis, and most (90%) are living in Brazil. According to the World Health Organization, individuals travelling to Ecuador, Colombia, Brazil, Guatemala, Mexico, and Venezuela are at a risk of onchocerciasis as well as infecting approximately 12.6 million individuals with lymphatic filariasis (80% in Haiti). Significant mortality and morbidity can be observed in cases where necessary precautions are not taken in individuals travelling to these regions and where appropriate prophylactic drugs are not administered.

Acanthamoeba is a free-living amoeba which can be isolated from environment and among others well known as an opportunist protozoan parasite causing infections in humans and animals. Eyes are extremely important for the wild birds and losing sight ability due to Acanthamoeba can be dangerous. The studies on Acanthamoeba infection in wild birds is very few in world and Turkey therefore we aimed to screen deceased wild birds found in İzmir and Manisa provinces located in western Turkey using PCR and non-nutrition agar (NNA) plate method. Cornea samples were obtained from 18 deceased wild birds. During the external examination, signs of keratitis were observed in two Eurasian sparrowhawks (Accipiter nisus). All of the corneal samples were analyzed by two PCR methods and NNA plate. According to results, the Acanthamoeba positivity in corneal samples was 16.6% and 5.5% by PCR and plate method, respectively. According to sequencing data, two of isolates belonged to genotype T5 and one was genotype T4. In conclusion, Acanthamoeba infection was detected in wild bird cornea samples with/without keratitis for the first time in the world. The result of this study also show that Acanthamoeba can be a cause of keratitis in wild birds of Turkey and thus these predator birds can be a target of other wild animals due to loss of sight ability. In terms of public health, these results show the importance of wild birds as a source of Acanthamoeba infection in nature.

Acanthamoeba is one of the most common free-living amoebas (FLA) that present in environment. In humans, Acanthamoeba can cause an infection of the eye termed Acanthamoeba keratitis, which mostly occurs in contact lens wearers. In the present study, we aimed to screen the presence of Acanthamoeba DNA in stray dogs using previously collected conjunctival swab samples in a hyper-endemic area for canine leishmaniasis. Totally, 184 dogs were included in the study and 27 of them (14.6%) were found positive for Acanthamoeba according to the 18s rRNA gene sequencing. Two different genotypes (T4 and T5) were identified and T5 was firstly reported in Turkey in the present study. Statistical analysis was performed and no correlation was found between Leishmania and Acanthamoeba positivity (P<0.05). To best of our knowledge, this is the first study conducted to screen Acanthamoeba among stray dogs. Further studies are necessary to reveal the infection status and genotypes among dogs and its possible correlation with leishmaniasis.

Background: The aims of this study were to identify Blastocystis subtypes (STs) in a cohort of Turkish patients with various gastrointestinal symptoms using a novel Real Time PCR method developed recently for Blastocystis detection and assess the relationship between Blastocystis STs and patient symptoms. Methods: Totally, 617 stool samples of patients with gastrointestinal symptoms were examined with microscopy and inoculated in Jones medium. Blastocystispositive samples were further assessed to identify coinfections with other possible pathogens, including bacteria and viruses. Diagnostic efficacies of microscopy,culture and Real-Time PCR were compared. PCR products were sequenced to identify the subtypes of Blastocystis isolates. Results: Totally 94 (15.24%) samples were positive for Blastocystis after all methods.Among these, 83 of 94 (88.3%) samples were identified with all methods,while 11 were positive only with Real Time PCR. Diarrhea and abdominal pain were the leading symptoms in the patients. The only pathogenic agent identified in 76 of 94 (80.9%) patients was Blastocystis. Subtype 3 was the leading Blastocystis subtype (44.6%), while subtypes 6 and 7 were firstly isolated from symptomatic patients in our region. Conclusion: Comparison of three diagnostic methods indicated Real Time PCR as the most sensitive and specific method. Blastocystis was the only pathogenic agent among symptomatic patients, with subtype 3 being predominant. Patients with subtypes 6 and 7 need further assessments concerning the zoonotic potential of Blastocystis.

Cyclospora cayetanensis and Cryptosporidium spp are the two emerging coccidian parasites causing human infections by inhabiting the intestinal tract and can either remain asymptomatically, or can cause endemic and epidemic diarrhea in both children and adults. In the present study, the prevalence of C. cayetanensis and Cryptosporidium spp were assessed in an epidemiological study in Izmir province, along with risk factors related to infection. A total of 873 individuals from both the city centre and counties were included in the study, and stool samples were examined with formalin ethyl acetate and Kinyoun acid-fast stained preparations. The prevalence of C. cayetanensis and Cryptosporidium spp were found to be 3.0% and 0.8%, respectively. Lower socio-economic conditions, lack of health insurance, consumption of tap water, eating in common places were all found to be associated with higher positivity for C. cayetanensis and Cryptosporidium spp. Abdominal pain was the prominent complaint of infected individuals. Despite relatively low prevalence rates, the presence of C. cayetanensis and Cryptosporidium spp in Izmir province requires further assessments of intestinal parasites. Public health measures that may prevent the local risk factors of infection should be improved and implemented.