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

The Prevention Of The Development Of Peritonitis With Different Disinfectants To Compare The Efficacy Of Gastric Lavage At The Notes

In the implementation of natural orifice transluminal endoscopic surgery ( NOTES) there is a risk of infection arising from the orifices of the body and the endoscope used. This study investigates the need for and efficiency of gastric lavage in reducing the risk of peritoneal infection in transgastric intraperitoneal surgery. In this study, Wistar-Albino rats were divided into 4 groups of 8. Gastric lavage was performed with three, but it was not performed on group 4. Blood samples were taken on the first and 14th days postoperatively, and a leukocyte count was made and C-reactive protein was measured. On the 14th postoperative day, the rats were sacrificed; aerobic and anaerobic culture specimens were taken from the peritoneal cavity, and biopsy samples were taken. Pathological examination of biopsy lavage in those groups, all for less inflammatory reaction and foreign body giant cell formation mikroabses were observed. In the all groups Leukocyte count and CRP measurements according to measurement values are rising. Culture examination of samples of seven different rats, while producing, anaerobic culture was not isolated in. As a result of gastric lavage should be done to ensure that surgery more safely performed of transgastric peritoneoscopy. There is no difference between the disinfectant solutions used, and the use of normal saline for gastric lavage is sufficientWo

PROTECTIVE EFFECTS OF GINKGO BILOBA EXTRACT AGAINST CISPLATIN OTOTOXICITY

Cisplatin is one of the most common agents employed in standard treatments of a variety of malignant tumors. However, its clinical application is limited because of serious and sometimes irreversible side effects, that include ototoxicity. Upon cisplatin treatment, several areas of the cochlea are damaged, including outer hair cells in the organ of Corti, spiral ganglion and the stria vascularis. Notwithstanding extensive research, the presently available treatments to prevent ototoxicity are scarcely effective. Cisplatin is thought to interfere with production of endogenous antioxidants that protect inner ear against reactive oxygen species (ROS). Outer hair cells are the most sensitive to ROS damage which can lead to apoptosis: strategies of chemoprevention include the administration of antioxidants to protect hair cells at an early stage in the ototoxic pathways. In this study we evaluated the protective effect of a nutraceutical product compounds with antioxidant activity (ACUVAL®, Scharper Healthcare, Italy). Ginkgo biloba is known for antioxidant properties and for this reason we tested by cytofluorimetry the otoprotective effects of a Ginkgo biloba extract (Ginkgoselect®) against cisplatin-induced toxicity in a mouse inner ear cell line (OCk3). The results support the hypothesis that the pre-treatment with Ginkgo biloba extract (50-100 µg/ml) is able to protect OCk3 against cisplatin-induced toxicity. We are presently investigating the effects of the same extract in rat inner ear organ cultures