Determination of the anti-protozoal activity of medicinal agents using the phenomenon of plaque formation by Acanthamoeba spp.

Representatives of the genus Acanthamoeba are among the most widespread protists in the environment. They have a ubiquitous distribution and can sometimes cause quite serious pathologies in humans. The treatment ofp rotozoal infections caused by free-living amoebae is currently limited and often unsuccessful. In the presented investigation, amebicidal activity was determined against both the trophozoites and cysts of Acanthamoeba spp., which were isolated during the microbiological examination of environmental objects. The inhibitory activity of drugs in vitro was determined using the authors' proposed method, which is based on the plaque formation phenomenon: this is initiated by free-living amoebae when cultured in agar containing the bacteria Cellulosimicrobium sp. strain bent-1. Based on a series of experimental studies, the paper proposes a reliable and inexpensive method for determining the anti-protozoal activity of medicinal agents, which will significantly complement the current screening method system when studying existing drugs, or new drugs during their development stage.

Research of biological properties of enterovirus strains associated with ischemic stroke.

OBJECTIVE: Introduction: The research of biological properties of enteroviruses associated with ischemic stroke (IS) allows us to identify their intratypic differences. The aim: to identify genetic markers of strains of enteroviruses associated with IS. PATIENTS AND METHODS: Materials and methods: 11 strains of enteroviruses isolated from the serum of patients with IS were identified in the virus neutralization test. Genetic markers of isolated strains (Abent, marker S, marker rct40) were determined. RESULTS: Results: Eleven strains of enteroviruses were isolated from the serum of patients with IS. Eight viruses: Coxsackie B viruses (serotypes 2, 3, 4) and ECHO viruses (serotypes 6, 9, 27 (two strains), 29) were identified in these strains. Other three strains of enteroviruses were unidentified. Different combinations of genetic markers were found. Seven strains of enteroviruses (Coxsackie B2, B3, ECHO 6, ECHO 9, ECHO 27 (two strains) and one unidentified virus) had virulence markers: Abent-, rct40+ and S-. Three strains (Coxsackie B4, ECHO 29, one unidentified virus) had markers: Abent-, rct40+, S+. Another one unidentified virus had markers: Abent+, rct40+, S -. CONCLUSION: Conclusions: All 11 isolates of enteroviruses associated with IS had rct40+ marker, 10 of the 11 isolates had marker Abent- and 8 of 11 isolates had marker S-. The research of genetic markers allows to perform typic and intratypic differentiation of strains of enteroviruses associated with the IS.

Applying of bacteria Cellulosimicrobium sp. for cultivation protozoa of genus Acanthamoeba

The aim of this study was identification features of cultivation representatives of genus Acanthamoeba isolated from bentonite using Cellulosimicrobium sp. as a bacteria-feeders. Identification of isolated bacteria was conducted by morphological, cultural and molecular-genetic methods. The cultivation of free-living "bentonite" amoeba on the lawn of Cellulosimicrobium sp. have gained significant advantages than using Escherichia coli as a bacteriafeeders was shown. "Bentonite" amoeba form crateroid plaques, which fit to the quantitative characteristic materials which contains amoeba, during deep co-cultivation Acanthamoeba sp. and Cellulosimicrobium sp. on 1% glucose meet-peptone agar.

Morphological, physiological and genetic characteristics of protozoa of genus Acanthamoeba, isolated from different deposit of bentonite in Ukraine

The representatives of genus Acanthamoeba are widespread in the environment. The presence of freeliving Acanthamoeba sp. in such mineral deposits as bentonite was shown for the first time. Identification of isolated amoeba was conducted according to morphological features of trophozoites and cysts, as well as using sequencing of gene 18S RNA (amplifier GTSA.B1). The obtained data showed that isolated amoebae belong to the genotype T4 and II morphological group (cyst size <18 µm). For its growth, "bentonite" amoebae are intensively used bacteria of the genus Cellulosimicrobium sp. as a food substrate.

The role of the persistent enterovirus infection in development of acute stroke.

INTRODUCTION: The role of enteroviruses in development of dilated cardiomyopathy, myocardial infarction, myocarditis, pericarditis is known. THE AIM: To examine the role of chronic enterovirus infections in development of acute stroke. MATERIAL AND METHODS: Blood samples from 72 patients with acute stroke (study group) and 35 patients without vascular disease (control group) were investigated by reverse transcription polymerase chain reaction for the presence of enterovirus RNA, by using virological method to detect enteroviruses, by ELISA for the levels of IgM and IgG antibodies to enteroviruses. RESULTS: The enteroviruses genomes were detected significantly often in the serum of patients with stroke (23,6 ± 5,9%) than in control group (2,9 ± 2,8%). The viruses were isolated and were identified as Coxsackie B (serotypes 2, 3, 4) and ECHO (serotypes 6, 9, 27 (two strains), 29), three strains have not been identified in study group. IgM to enteroviruses were not found in the sera of both groups of patients. IgG to enteroviruses were detected in 17 patients in study group (23,6 ± 5,9%) and 2 patients in control group (5,7 ± 3,9%). The presence of enteroviruses genomes and IgG in sera of patients in control group (11,1 ± 3,7%) indicate the persistence of enteroviruses. The proportion of patients with IgG to enteroviruses in sera is higher in study group (12,5 ± 3,9%) than in control group (5,7 ± 3,9%). CONCLUSION: The enterovirus infections play trigger role in development of acute stroke.