Determination of Diversity of Actinomycete in Forest Soil Subject To Different Blood Groups; Classical Approaches Before Metagenomic

Objective: Forensic microbiology is the developing field of forensic science. Forensic microbiologist can use trace evidence to match people to crime scenes, to investigate bioterrorism incidents, and to determine cause and time of death. In recent years, important studies on the environmental microbiota have been carried out. These studies can be associated with many legally valuable data as well as being related to the microbiota of environmental events. Actinobacteria are gram-positive micelle bacteria with enormous diversity, common in soil, and have high economic value. So far 47 families and 711 species of Actinobacteria were isolated and identified in variety of habitats. Increasing forensic studies in recent years show that microbial profiles can be used as evidence. Methods: In this study, actinobacteria were isolated from forest soil samples exposed to eight different blood groups for two weeks and control group. Genomic DNA isolation from colonies selected by considering comparative colony morphology, PCR amplification targetted 16S rRNA and sequence analysis were carried. Results: A total of five Actinobacteria type bacteria (three Micromonospora sp., one is Streptomyces sp., and one Actinomadura sp.) were obtained from the soil samples mixed with the B Rh (+) blood group in the 7th day. No Actinobacteria growth was observed neither in the control group nor in the soils to which other blood groups were added. According to the p-distance values of all isolates, they were determined as subspecies rather than a new species. Conclusion: This is a preliminary study to identify bacterial communities that may be present and differ in soil exposed to blood. The inability to isolate Actinobacteria from most of the soil samples may be due to the chemical or enzymatical properties of the bloods that can degrade the bacterial spores. In the further studies, different media and genomic techniques should be tried in different types of soils. © (2023), (Refik Saydam National Public Health Agency (RSNPHA)). All Rights Reserved.FBE19004.20.011The authors wish to thank Başkent University Head of Department Prof. Dr. Özlem Darcansoy İşeri and all BU Molecular Biology and Genetics Department faculty members and assistants. We would also like to thank Hitit University Scientific Research Projects Coordinator for their support within the scope of the project numbered FBE19004.20.011

Detection of Blastocystis Subtypes in Children With Functional Abdominal Pain and Celiac Disease in Çorum, Turkey

Background: Blastocystis has been associated with various symptoms of the gastrointestinal tract. We aimed to investigate the prevalence of Blastocystis in children with celiac disease (CeD) or functional abdominal pain (FAP) and to evaluate its subtypes (STs) with respect to demographic, socioeconomic and epidemiological factors. Methods: Overall, 161 fecal samples were collected from healthy children and patients with FAP or CeD in Hitit University Erol Olçok Research and Training Hospital, Corum, Turkey between 2016-2018. Samples were examined using both native-Lugol (NL) and trichrome-stained (TS) smears, and further analyses by PCR and Sanger sequencing were performed. A standard questionnaire was applied to obtain demographic, socioeconomic, epidemiological data. Results: Blastocystis was found in 10.6% of the total study population. Neither bacteria nor any other parasites were found, except for one Giardia (0.6%) in the CeD group. The presence/absence of the parasite was not found to be associated with demographic, socioeconomic and epidemiological factors. Blastocysis was detected in 11.5% (6/52) of the CeD, 7.7% (4/52) of the FAP, and 12.3% (7/57) of the healthy group. Diagnostic methods were similar in terms of Blastocystis detection (P= 0.671), and there was fair agreement between the NL, TS and PCR (Fleiss' Kappa=0.847, P=0.001). ST2 (42.8%) and ST3 (35.7%) were the predominant STs followed by ST1 (21.4%). Conclusion: We observed no difference between study groups in terms of Blastocystis prevalence. ST1, ST2 and ST3 subtypes were detected. Blastocystis prevalence and STs were not related to any of the demographic, socioeconomic and epidemiological factors. © 2022 Güreser et al. Published by Tehran University of Medical Sciences.Funded by Hitit University Scientific Research Projects Unit, Grant/Award Number: TIP19002.15.007TIP19002.15.00

Prevalence of Blastocystis and Dientamoeba Fragilis in Diarrheal Patients in Corum, Türkiye

Article; Early AccessTo investigate the prevalence of Blastocystis and Dientamoeba fragilis in diarrhea patients and healthy individuals in Corum, Türkiye, fecal samples from 92 diarrhea patients and 50 healthy individuals were collected and evaluated using direct microscopy and molecular methods to screen for bacteria, protozoa, and viruses. The prevalence of Blastocystis was 24.6% in total and more frequent in the healthy group (30.0%). The commonly detected STs (subtypes) were ST3 (40.0%) and ST2 (34.2%). The distribution of Blastocystis STs in the healthy and diarrheal groups did not show any difference in sex and age, but ST3 was detected more frequently in patients aged from 40 to 59 years (p 0.05). Alleles 4 (8/12) and 2 (4/12) were present in ST1; 9 (3/5) and 12 (2/5) in ST2; 34 (9/14), 36 (3/14), and 38 (2/14) in ST3; and only allele 42 (2/2) in ST4. D. fragilis was present in 8.4% of the population. However, there was no statistically significant difference between the healthy and diarrheic groups (12.0% and 6.5%, respectively), neither with respect to age nor sex. Co-infection was 58.3% and was more frequent in healthy individuals (33.3%) than in diarrhea patients (25.0%). Blastocystis ST3 was the most common subtype detected, with D. fragilis at 33.3%. Salmonella, Shigella, or helminth eggs were not observed in all groups, but Entamoeba histolytica, Giardia intestinalis, Cryptosporidium, Rotavirus, Adenovirus, and Clostridium difficile toxin were found only in diarrhea patients. These findings support the hypothesis that Blastocystis and D. fragilis may be part of the healthy human gut microbiome. © 2023, The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.We thank Hitit University Scientific Research Projects Commission (Grant No: TIP19001.16.005), Gazi University Scientific Research Unit (Grant No: 01/2017-15), and TOBB University of Economics and Technology for their financial support.TIP19001.16.005; Gazi Üniversitesi: 01/2017-1

Reduced Phosphorylated Foxp3 Levels in Crimean Congo Haemorrhagic Fever

Crimean-Congo haemorrhagic fever (CCHF) is a severe human infection which can lead to fatal consequences. Acute CCHF patients were previously shown to exhibit frequencies of regulatory T-cell (Treg) but lower Treg-mediated suppressive activities than the healthy counterparts. This study aims is to investigate the phosphorylation levels of Foxp3 protein (master regulator of Treg cells) in CCHF patients. Blood samples collected from 18 CCHF patients and nine healthy volunteers were used to isolate peripheral blood mononuclear cells (PBMCs). Total and phosphorylated Foxp3 expression levels in the isolated PBMC samples were monitored by western blot and quantified using ImageJ software. Total Foxp3 expression levels in CCHF patients displayed decreasing trend, but not significantly. In contrast, significantly lower expression levels of phosphorylated Foxp3 were reported in CCHF patients. Our results suggest a possible association between Foxp3 dephosphorylation and CCHF pathogenesis. Nevertheless, more studies are required to evaluate the effect of Foxp3 dephosphorylation on Treg function, which would not only help to enlighten the CCHF pathogenesis but also contribute to the development of effective treatment strategies.We thank to Atlas Biotechnologies (Ankara/Turkey) for the technical support and TOBB Technology and Economics University/Ankara/Turkey for the financial funding (08.11.2021-DAF-2021-6).TOBB Technology and Economics University/Ankara/Turkey [08.11.2021-DAF-2021-6