Detection of enteric parasites and molecular characterization of Giardia duodenalis and Blastocystis sp. in patients admitted to hospital in Ankara, Turkey.

This epidemiological study assesses the occurrence of enteric parasites in 4303 patients attended at two public hospitals in Ankara (Turkey) during 2018-2019. Microscopy was used as a screening test. Giardia duodenalis was also identified using a commercial ELISA for the detection of parasite-specific coproantigens. Giardia-positive samples by microscopy/ELISA were confirmed by real-time PCR and characterized using a multilocus genotyping scheme. Blastocystis sp. was genotyped in a sample subset. Blastocystis sp. (11.1%, 95% CI 11.4‒14.8%) and G. duodenalis (1.56%, 95% CI 1.22‒1.96) were the most prevalent pathogens found. Cryptosporidium spp., Entamoeba histolytica and intestinal helminths were only sporadically (<0.5%) found. For G. duodenalis, sequence (n = 30) analyses revealed the presence of sub-assemblages AII (23.3%), discordant AII/AIII (23.3%) and mixed AII + AIII (6.7%) within assemblage A, and BIII (10.0%), BIV (3.3%) and discordant BIII/BIV (23.3%) within assemblage B. Two additional sequences (6.7%) were assigned to the latter assemblage but sub-assemblage information was unknown. No associations between G. duodenalis assemblages/sub-assemblages and sociodemographic and clinical variables could be demonstrated. For Blastocystis sp., sequence (n = 6) analyses identified subtypes ST1, ST2 and ST3 at equal proportions. This is the first molecular characterization of G. duodenalis based on MLG conducted in Turkey to date.This research was partially funded by the Scientific Research Unit of Gazi University (Ankara, Turkey) under project number 01/2017-15, and by the Health Institute Carlos III (ISCIII), Ministry of Science, Innovation and Universities (Spain) under grant number PI16CIII/00024N

Blastocystis: A mysterious member of the gut microbiome

Blastocystis is the most common gastrointestinal protist found in humans and animals. Although the clinical significance of Blastocystis remains unclear, the organism is increasingly being viewed as a commensal member of the gut microbiome. However, its impact on the microbiome is still being debated. It is unclear whether Blastocystis promotes a healthy gut and microbiome directly or whether it is more likely to colonize and persist in a healthy gut environment. In healthy people, Blastocystis is frequently associated with increased bacterial diversity and significant differences in the gut microbiome. Based on current knowledge, it is not possible to determine whether differences in the gut microbiome are the cause or result of Blastocystis colonization. Although it is possible that some aspects of this eukaryote’s role in the intestinal microbiome remain unknown and that its effects vary, possibly due to subtype and intra-subtype variations and immune modulation, more research is needed to characterize these mechanisms in greater detail. This review covers recent findings on the effects of Blastocystis in the gut microbiome and immune modulation, its impact on the microbiome in autoimmune diseases, whether Blastocystis has a role like bacteria in the gut–brain axis, and its relationship with probiotics

Comparison of Methods for Detection of Blastocystis Infection in Routinely Submitted Stool Samples, and also in IBS/IBD Patients in Ankara, Turkey

BACKGROUND: This study compared diagnostic methods for identifying Blastocystis in stool samples, and evaluated the frequency of detection of Blastocystis in patients with irritable bowel syndrome (IBS) and inflammatory bowel disease (IBD). RESULTS AND DISCUSSION: From a set of 105 stool specimens submitted for routine parasitological analysis, 30 were identified as positive for Blastocystis by the culture method. From that group of 30 positives, Lugol's stain, trichrome staining, and an immunofluorescence assay identified 11, 15, and 26 samples as positive respectively. Using culture as a standard, the sensitivity of Lugol's stain was 36.7%, trichrome staining was 50%, and the IFA stain was 86.7%. The specificity of Lugol's stain was 91%, trichrome staining was 100%, and the IFA stain was 97.3%. In the group of 27 IBS and IBD patients, using all methods combined, we detected Blastocystis in 67% (18/27) of the patients. Blastocystis was detected in 33% (2/6) of IBD patients and 76% (16/21) of IBS patients. For comparison, trichrome staining alone, the method most frequently used in many countries, would have only identified Blastocystis infection in 29% (6/21) of the IBS patients. No parasitic co-infections were identified in the IBS/IBD patients. Most Blastocystis-positive IBS/IBD patients were over 36 with an average length of illness of 4.9 years. CONCLUSIONS: Most IBS patients in this study were infected with Blastocystis. IFA staining may be a useful alternative to stool culture, especially if stool specimens have been chemically preserved

The effect of two artificial salivas on the adhesion of Candida albicans to heat-polymerized acrylic resin.

Xerostomia can diminish the quality of life, leads to changes in normal chemical composition of saliva and oral microbiata, and increases the risk for opportunistic infections, such as Candida albicans. Various artificial salivas have been considered for patients with xerostomia. However, the knowledge on the antifungal and antiadhesive activity of artificial saliva substitutes is limited. The aim of the present study was to evaluate influence of two artificial salivas on the adhesion of Candida albicans to the polymethylmethacrylate disc specimens

The effect of two artificial salivas on the adhesion of Candida albicans to heat-polymerized acrylic resin

PURPOSE. Xerostomia, can diminish the quality of life, leads to changes in normal chemical composition of saliva and oral microbiata, and increases the risk for opportunistic infections, such as Candida albicans. Various artificial salivas have been considered for patients With xerostomia. However, the knowledge on the antifungal and antiadhesive activity of artificial saliva substitutes is limited. The aim of the present study was to evaluate influence of two artificial salivas on the adhesion of Candida albicans to the polymethylmethacrylate disc specimens. MATERIALS AND METHODS. Two commercial artificial salivas (Saliva Orthana and Biotene Oral Balance Gel) Were selected. 45 polymethylmethacrylate disc specimens were prepared and randomly allocated into 3 groups; saliva Orthana, Biotene-Oral Balance gel and distilled water. Specimens were stored in the artificial saliva or in the sterile distilled water for 60 minutes at 37 degrees C. Then they were exposed to yeast suspensions including Candida albicans. Yeast cells were counted using x40 magnification under-alight Microscope and data were analysed. RESULTS. Analysis of data indicated statistically significant difference in,adhesion of Candida albicans among all experimental groups (P=.000). Findings indicated that Saliva Orthana had higher adhesion scores than the Biotene Oral Balance gel and distilled water (P<.05). CONCLUSION. In comparison of Saliva Orthana, the use of Biotene Oral Balance Gel including lysozyme, lactoferrin and peroxidase may be an appropriate treatment method to prevent of adhesion of Candida albicans and related infections in patients with xerostomia

Investigation of the Presence of Blastocystis spp. in Stool Samples with Microscopic, Culture and Molecular Methods

Blastocystis species are enteric protozoa frequently detected in human and animals. Seventeen subtypes (STs) have now been identified, nine of them isolating from humans. The pleomorphic structure and genetic diversity of Blastocystis spp. and the absence of standardized diagnostic methods complicate the evaluation of current data. Microscopic methods such as native-lugol and trichrome staining are most frequently used methods in routine diagnosis, while culture and molecular methods are preferred for research purposes. The aims of this study were to investigate the presence of Blastocystis spp. in the stool samples of patients with gastrointestinal complaints by microscopic and culture methods, and to detect the subtypes of isolates by polymerase chain reaction (PCR). A total of 350 stool samples collected from patients with diarrhea (n= 157) and without diarrhea (n= 193) were included in the study. Presence of Blastocystis spp. in the samples were investigated by native-lugol examination, trichrome staining and direct fluorescent antibody (DFA) methods. Ringer's solution containing 10\% horse serum and 0.05\% asparagine was used for cultivation. The cultures were evaluated after 3-4 days of incubation at 37 degrees C by microscopic examination. The subtypes of Blastocystis spp. strains isolated from the cultures have been identified by PCR using sequence-tagged site primers. A total of 66 (19\%) stool samples, of them 26 (16.6\%) were from diarrheal and 40 (21\%) from non-diarrheal cases, yielded Blastocystis sp. growth in culture. Among the evaluated samples, 12\% (42/350) were found positive with native-lugol examination, 17\% (58/350) with trichrome staining, and 19\% (66/350) with DFA method. The agreement of culture and native-lugol method was estimated as strong (kappa= 0.752), while it was very strong between culture with trichrome staining and DFA methods (kappa= 0.922 and kappa= 1.00, respectively). When the culture was accepted as reference method, the sensitivity and specificity rates of native-lugol method were 65\% and 100\%, trichrome staining method were 88\% and 100\%, and DFA method were 100\% and 100\%, respectively. Forty-three (65\%) of Blastocystis spp. positive samples were subtyped by PCR, while 23 isolates could not be subtyped. The most frequent detected subtype was ST3 (12/43; 28\%), followed by ST1 (6/43; 13.9\%), ST4 (5/43; 11.6\%) and ST7 (5/43; 11.6\%), ST2 (3/43; 7\%) and ST6 (1/43; 2.3\%). ST5 was not detected in this study and 11(25.6\%) samples have been identified to have mixed subtypes. The differences of Blastocystis spp. positivity rates and the distribution of the subtypes between the patients with or without diarrhea were not found statistically significant (p> 0.05). In our study, ST3 was the most frequently identified Blastocystis spp. subtype, similar to the previous national studies, however ST6 and ST7 have been identified for the first time. In conclusion, as the sensitivity of native-lugol examination is low, culture is time-consuming and laborious and PCR methods are costly and non-standardized, rapid, practical and high sensitive DFA is considered as the favourable method in the diagnosis of Blastocystis spp. in routine laboratories

Demodex spp. as a possible aetiopathogenic factor of acne and relation with acne severity and type

Introduction: Acne is a very common skin disease in adolescents and young adults, but it also affects adults. However, its aetiology is not yet fully understood. Demodex appears to be associated with multiple skin disorders, but controversy persists. Some reports indicate a connection between acne vulgaris and demodicosis. Aim: To confirm the association between Demodex infestation and acne vulgaris. Material and methods: A total of 108 patients were enrolled in the acne group. Acne severity was calculated as GASS and acne type (adolescent and post adolescent) was recorded. An age-sex matched healthy control group comprising 65 individuals were included in the study. Dermatological examinations were performed and an SSSB was used to determine the presence of Demodex. Results: In our study, Demodex positivity was seen in 46 (42.6\%) patients in the acne group and 8 (12.3\%) in the control group; this difference was statistically significant (p < 0.001). A multivariate Backward Step-By-Step Logistic Regression analysis identified the most effective factors for acne development such as Demodex positivity (OR = 5.565, 95\% CI: 2.384-12.99 and p < 0.001) and age under 25 years (OR = 2.3 and 95\% CI: 1.183-4.473 and p = 0.014). Alcohol consumption was related to Demodex positivity (p = 0.019) in post adolescent acne. Conclusions: Our study is the first one to evaluate acne severity, acne type and the relationship to Demodex prevalence. We suggest that Demodex infestation should be considered when the classical therapies are ineffective especially in cases of post adolescent acne

Second International Blastocystis Conference

En esta tercera parte de la conferencia 6 invitados exponen sus investigaciones comenzando con Christen Rune Stensvold, con su ponencia titulada Upgates on detection subtypes, host specificity and associations to microbiome signatures. Luego interviene Raul Tito Tadeo de Perú habla acerca de su experiencia con asociaciones con la microbiota intestinal en la gente de Bélgica; su presentación se titula Blastocystis: Beyond genus and its association with the human gut microbiota. En seguida participa John Anthony Yason National University of Sngapore Proyecto: A Potentially Pathogenic Blstocystis Subtype Disrupst Gut Microbiota. Luego interviene Simona Gabrielli Assistant Professor of Parasitology – Sapienza University of Rome, Italy; el título de su proyecto es “On the ocurrence of Blastocystis subtypes and their correlation with faecal microbiota in HIV patiens referres to the Umberto I University Hospital in Rome Italy. Continua Justinn Hamilton University of Copenhagen, Denmark, su ponencia se titula Exploring interactions between Blstocystis, other intestinal parasites end the gut microbiomes of wild chimpanzees in Senegal y finalmente, expone su proyecto Funda Dogruman-Al Associate Professor-Medical Faculty of Gazi University-Turkey su ponencia es Blastocystis in Clinical Microbiology: advantages and disadvantages of methods used in dignosis. What can we do for standardization

Late-stage systemic immune effectors in Plasmodium berghei ANKA infection: biopterin and oxidative stress

To investigate the involvement of systemic oxidative stress in the pathogenesis of murine cerebral malaria, mice were infected with the Plasmodium berghei (P. berghei) ANKA 6653 strain. Serum tryptophan (Trp), kynurenine and urinary biopterin, liver, brain, spleen and serum superoxide dismutase (SOD), glutathione peroxidase (GPx), malondialdehyde (MDA) and nitrite and nitrate (NOx) levels were measured on day 7 post-inoculation. Our data showed a significant decrease in SOD and an increase in GPx activity and MDA level in all the examined biological materials (p < 0.05), except spleen. Conversely, GPx activities in spleen were depleted, while SOD and MDA levels remained unchanged. Increased MDA levels might indicate increased peroxynitrite production, lipid peroxidation and oxidative stress. Also, elevated urinary biopterin, which was accompanied by increased NOx (p < 0.05), may support the inhibition of Trp degradation (p > 0.05). The excessive NO synthesis in P. berghei infection may be related to the up-regulation of inducible NO synthase, which was in accordance with the increased biopterin excretion. Thus, the large quantities of released toxic redox active radicals attack cell membranes and induce lipid peroxidation. Although P. berghei infection did not demonstrate systemic Trp degradation and related indoleamine-2,3-dioxygenase activity, it may cause multi-organ failure and death, owing to host-derived severe oxidative stress