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Introduction. Modern parasitology is an area of interdisciplinary research that uses the methods and approaches of various biological disciplines, such as genomics, proteomics, bioinformatics, ecology, biology, medicine, veterinary medicine, climatology, using methods of landscape-geographical mapping, mathematical modeling to assess the parasitological situation and predict its dynamics. New problems associated with the parasitic aspects of global climate change, the introduction of new host species and parasites (pathogens), and anthropogenic transformation of natural ecosystems become topical. In this regard, the primary tasks for optimizing the socio-hygienic monitoring of the parasitic safety of the environment is the introduction of new methodological approaches for ranking territories - the risk according to the species composition and intensity of circulation of pathogens of parasitic diseases, the trend in changes in their range of pathogens and epidemiological properties in anthropogenically determined conditions. The purpose of the study is to substantiate the methodology for choosing a territory for optimizing social and hygienic monitoring and substantiating proposals for the unification of hygienic standards for parasitological indicators of environmental safety. Materials and methods. Experimental parasitological monitoring was carried out at the pilot site. 712 samples were studied using unified methods of parasitological research of biomaterial (MUK 4.2.3145-13 “Laboratory diagnostics of helminthiases and protozooses”) and environmental objects (MUK 4.2.2661-10 “Methods of sanitary and parasitological research”). Results. Data were obtained on the high intensity of the circulation of pathogens of parasitic diseases in the environment - 25%. More than 15 types of pathogens of parasitic diseases have been systematized. Limitations. The limitations of the study were associated with the lack of development of diagnostic tools for the genetic identification of parasitic pathogens within the same systematic group by PCR analysis. Conclusion. Methodological disagreements in various normative documents to the composition of the “target indicators” of research are determined. The practical part of the work included the collection of material from people and animals, from environmental objects. A new approach to the hygienic standardization of parasitological indicators based on group systematization of the propagative stages of development of parasites with subsequent methodological determination of their taxonomic affiliation is substantiated.

Introduction. MALDI-TOF identification method is widely used in microbiology due to its accuracy and rapid results achievement. However, applying this method to mold fungi faces some difficulties and is not always effective. Purpose of the study. The aim of the study was to evaluate the profits of using the long cultivation and protein extraction protocol in routine identification of mold fungi isolates from environment. Materials and methods. The analysis of molds museum collection from Centre for Strategic Planning of FMBA of Russia was performed by MALDI-TOF mass-spectrometry Biotyper (Bruker Daltonics) with cultivation in liquid media and long optimized protein extraction protocol with acetonitrile and formic acid. Results. One hundred thirty seven isolates were analyzed. Quality spectra were achieved for 71.5% of samples. Identification with MBT Filamentous Fungi Library database with the high confidence score (> 1.7) was achieved for 55% of isolates (26% with score >2). Samples analyzed included members of nineteen families and 27 genera. 16% of samples were not identified despite producing high-quality spectra. Limitations. When studying the possibility of using the time-of-flight mass spectrometry method to identify mold fungi, a sample of 137 isolates of mold fungi from the environment was analyzed, which is a sufficient reference sample. The analyzed samples included representatives of 19 families and 27 genera, which makes it possible to apply the findings to at least these representatives of micellar fungi. In this study 22 samples with good quality spectra, were not identified with MBT Filamentous Fungi Library database. In the future studies, these samples, along with other samples like that, will be identified by genetic molecular methods and added to the new home-made database for filamentous fungi MALDI-TOF identification. Conclusion. Effective identification of filamentous fungi by mass-spectrometry methods requires pure culture achieved from liquid media, long optimized protocol of protein extraction and building an in-house database of spectra not presented in Bruker database.

Introduction. Even in the modern urban environment humans are in constant direct and indirect contact with soil. This leads to the spread of a wide range of soil-transmitted human and animal pathogens. Therefore, the development of fast and inexpensive methods of analysis and monitoring of these pathogenic objects is of great importance. PCR method is widely applied in laboratory practice and is able to detect even the uncultivated types of pathogens. The aim of the study was to optimize the method of DNA isolation from soil, making it suitable for PCR-assay. Materials and methods. DNA was isolated from the samples of surface layer of forest soil rich in humus, using lab-shelf chemicals and/or commercial kit. RT-PCR-test was performed using universal bacterial primers. Results. We have analyzed various combinations of four extraction methods and three pre- and post-treatment methods. DNA was efficiently extracted by all methods, however, without additional purification stages it was unsuitable for PCR. The calcium salts treatment ws demonstrated to be necessary for removal of PCR inhibitors, presumably humic acids. Two DNA isolation methods were developed. Both methods use incubation with CaCO3 suspension followed by cetrimonium bromide lysis. More sensitive and unexpensive method uses CaCl2 as an additional purification stage. The less sensitive but more reproducible method included DNA isolation on Qiagen DNA (Qiagen) columns. Limitations. When working out the technique of DNA isolation for PCR analysis, samples of the only sod-podzolic soil were studied. Therefore, the technique can be applied only for this type of soil. Conclusion. Both methods optimized in this study can be used for evaluation of soil samples for the presence of pathogens by PCR.

Introduction. The general issues of methodology, approaches and technologies in laboratory diagnostics of parasitic infections include MALDI-ToF MS-analysis (Matrix-Assisted Lazer Desorption/Ionization Time-of-Flight Mass Spectrometry; matrix-activated laser desorption-ionization time-of-Flight mass spectrometry), as well as a number of specific issues related to the relevance and timely application of this highly specific technologies in identification and typing of pathogens of parasitic diseases. An important and practically unsolved problem in parasitology is high-quality, highly effective, highly sensitive laboratory diagnostics of parasitoses. The object of this study is to analyze and evaluate the current state and perspectives of MALDI-TOF as an instrument of identification of diverse parasites in vples. Materials and methods. Federal statistics data on infectious and parasitic morbidity, own experimental studies, retrospectively analyzed literary data of foreign and Russian authors. Results. A steady decline in the detection of cases of parasitic invasions is associated with the lack of highly effective and sensitive diagnostic methods. Limitations. In the process of studying the effectiveness of the MALDI-TOF MS method in parasitological laboratories (departments), we analyzed our own experimental data (samples with artificial introduction of parasitic pathogens) and analyzed data on the main parasitic nosologies circulating in the territory of the Russian Federation for three years from 2017-2019, which are reflected in the state reports of of the Federal Service for Supervision in Protection of the Rights of Consumer and Man Wellbeing, which is a fairly reference sample. Conclusions. Studies comparing the results of mass spectrometric typing and traditional diagnostic methods are promising, which are necessary to clarify the capabilities of the method and determine its place in the laboratory diagnosis of infections caused by parasitic pathogens.

Introduction. Pathogenic Escherichia coli present a real threat to human health. One of the ways of transmission of these isolates is via environmental water sources. Therefore, evaluation of pathogenic potential of E. coli population in water is of great interest. Purpose of the study. The aim of this study was to compare E. coli isolates from wells, sewers, water pools and surface waters were compared with two control groups - “non-pathogenic” isolates from feces of healthy people and “potentially pathogenic” from feces of people with inflammatory bowel diseases (IBD). Materials and methods. PCR-assay was used to detect potential virulence genes. 19 E. coli virulence genes were analyzed: 11 toxins, 5 adhesion and invasion proteins and 2 diarrhogenic serotypes. The PCR identification of carbapenemase genes and various E. coli pathotypes was performed with the commercial “Amplisense” kits according to the manufacturer’s instruction. The assay was performed on 47 E. coli isolates from water environmental sources (WES), 44 isolates from feces of “practically healthy” people, 43 isolates from feces from IBD patients. Results. Isolates from WES were found to be similar to the group of isolates from healthy people. Only 2 types of virulence E. coli were detected in these groups - toxins CNF1 and 2 and invasin einv. IBD group of isolates demonstrated striking difference from the others. Only IBD isolates demonstrated such genes as adhesion regulator aggR, invasive antigen ipaH, hemolysin hly and antibiotic resistance gene NDM. CNF1 gene was found in IBD group significantly more often, than in two other groups. The only pathotype detected in the samples analyzed, enteroaggregative, was limited to the IBD group, too. Limitations. To compare the pathogenetic potential of E. coli from human feces and environment, 134 isolates were tested for 19 pathogenic genetic determinants, which is a representative selection. Within the analysis, we were unable to compare bacterial pathogenic potential from various environmental sources (surface waters and sewage, treatment facilities etc.) due to the uneven representation of these objects in the selection. It will be the subject of our future studies. Conclusion. Pathogenic potential of E. coli isolates from environmental water sources was close to that from healthy human feces.

Introduction. The processes of socio-economic restructuring in recent decades, taking place in our country, have led to a reduction in the volume and deterioration in the quality of measures to protect various components of the human environment from contamination by parasitosis pathogens. The number of diagnostic examinations of the population to identify patients with parasitic diseases has decreased. Every year, the interest in this problem of the medical and scientific community increases in the scientific literature of both foreign and Russian authors. There are more and more new prerequisites for studying the interaction of the intestinal microflora, parasites, the immune response inflammatory processes of the human body. Today, the role of parasitic intestinal protozoa Blastocystis spp., and in some cases aggravated by the addition of opportunistic fungi of the genus Candida spp., is increasingly being discussed in human pathology. The aim of the study was to assess the possible relationship between the presence of blastocyst invasion in the examined group of “practically healthy people” with various abnormalities in the gastrointestinal tract (diarrhoea, constipation, single or repeated pain, etc.), immunopathological processes (skin rashes, intolerance to any food, etc.), as well as the addition of opportunistic fungal microflora of the genus Candida spp. Materials and methods. A complex of parasitological and additional research methods was used, including a culture method for isolating fungi of the Candida spp. A complete analysis of the personal data received from the analyzed group was carried out. Results. The results of studies of the intestine’s clinical material (faeces) and the analysis of personal data in the surveyed “conditionally healthy” group of respondents confirmed our assumption about the pathological effect of blastocyst invasion on the conditionally healthy intestinal microflora and, accordingly, human health. Limitations. The limitations of this study make it possible to apply the findings to the health of the adult, non-old, able-bodied population and, at the same time, take into account regional characteristics in the development of mixed infections of parasitic and fungal genesis in humans. In this regard, we consider it possible to apply the findings to all regions with similar living conditions (urban lifestyle, cold and temperate climate, etc.) Conclusions. It is necessary to continue conducting further observations and research of various groups (individuals) in various regions of the Russian Federation, participating in the development of educational and practical algorithms for laboratory diagnostics of parasitoses and mycoses for specialists in clinical laboratories.

Introduction. The propagation of multi-resistance to antibiotics among hospital isolates of Klebsiella pneumoniae (K. pneumoniae) is a subject of growing concern worldwide. At present, growing data of association between resistance and hypervirulence in clinical isolates of K. pneumoniae emerges. However, the occurrence of these pathogens in the environment remains an open question. The aim of this study was to evaluate and compare antibiotic resistance determinants occurrence in Klebsiella pneumoniae isolates from water sources (environmental and sewage), human sources (practically healthy people and patients with inflaammatory bowel disease (IBD), and extraintestinal infections (ExII)). Materials and methods. The PCR assay of carbapenemase genes IMP, NDM, VIM, KPC, OXA-48 was performed with the commercial “Amplisense” kits according to the manufacturer's instructions. The assay was used to evaluate the occurrence of antibiotic-resistance genes in 223 isolates of Klebsiella pneumoniae from various sources: 42 isolates from sewage, 19 isolates from surface water sources, 30 isolates from biological material (blood, urine, surgical wounds, bronchoalveolar lavage) of patients with extraintestinal infections (ExII), 69 isolates from patients with inflammatory bowel diseases (IBD), and 63 isolates from faeces of practically healthy people. Results. The ExII group revealed various antibiotic resistance genes. The most prevalent gene was OXA (30% had this gene only, other 26,6% had also KPC or NDM). NDM as the only resistance gene was observed in 23,3% of ExII isolates. KPC gene was observed in 3,3% of ExII group. Two isolates from IBD group contained NDM gene along with VIM gene. Only NDM gene was found in all the other groups of Klebsiella pneumoniae isolates (13-28% isolates in every group, no statistical difference). NDM was shown to be associated with virulence genes iutA and rmpA that are responsible for iron consumption and hypermucoid phenotype. Conclusion. The most abundant resistance genes in the studied Klebsiella pneumoniae isolates were NDM (13.5%) and OXA (8%). At the same time, NDM was the only gene found in all groups (11-28%). NDM metallobeta-lactamase gene was associated with rmpA and iutA genes, giving an example of the connection between virulence and resistance properties. A significant amount of resistant isolates from healthy donors and surface waters indicates the need for additional study of the role of NDM positive isolates in pathogenicity of Klebsiella pneumoniae.

Conduction. The recognition diagnosis of helminths and protozoa invasion is one of the current and almost unresolved problems in modern parasitology. For the present, the lack of a standardized museum of parasitic pathogens in Russia and abroad complicates a parasitologist’s already hard work. The population urbanization, increased anthropogenic pressure, climate change, and natural disasters lead to the occurrence of favorable conditions for the development, long-term survival, and spread of pathogens of parasitic diseases in various environmental objects. The deterioration of the population socio-economic conditions, the emergence of interethnic conflicts in the world led to large-scale migration flows from countries near and far abroad to Russia, which led to an increase in pathogens circulating in the territory of the Russian Federation. The aim of our research was to improve the methodological support and create virtual slides with the image of larvae and eggs of helminths, as well as cysts and oocysts of pathogenic protozoa recorded on the territory of the Russian Federation based on digital scanner microscopes. Material and methods. The authors used scanner microscopes with the software module “Parasitology.” Results. An up-to-date register of photo images of more than 40 species of parasitic pathogens of medical significance and 53 species of pathogens of parasitic diseases of animals that pose a danger to human life has been delivered. Discussion. The authors are always working on improving the technical characteristics of digital analyzers. Conclusion. Updating the atlas, creating a new advanced gallery of helminth eggs, larvae, cysts, and oocysts of pathogenic protozoa will significantly increase parasitological studies’ effectiveness in sanitation and the analysis of intestinal microbiota.

Introduction. The purpose of our work was to justify the need to improve the legislative, regulatory and methodological framework and preventative measures in relation to the spread of parasitic infections in the provision of medical care. There is a wide range of pathogens of parasitic infestations that are transmitted to humans through various medical manipulations and interventions carried out in various medical institutions. Contaminated care items and furnishings, medical instruments and equipment, solutions for infusion therapy, medical personnel’s clothing and hands, reusable medical products, drinking water, bedding, suture and dressing materials can serve as a major factor in the spread of parasitic infections in the provision of medical care. Purpose of research is the study of the structure and SMP of parasitic origin, circulating on the objects of the production environment in multi-profile medical and preventive institutions of stationary type in order to prevent the occurrence of their spread within medical institutions. Material and methods. The material for the study was flushes taken from the production environment in 3 multi-profile treatment and prevention institutions of inpatient type: a multi-specialty hospital, a maternity hospital and a hospital specializing in the treatment of patients with intestinal diseases for the eggs of worms and cysts of pathogenic protozoa. Results. During the 2-year monitoring of medical preventive institutions, a landscape of parasitic contamination was found to be obtained from the flushes taken from the production environment objects in the premises surveyed as part of the research work. Discussions. In the course of research, the risk of developing ISMP of parasitic origin was found to be determined by the degree of epidemiological safety of the hospital environment, the number and invasiveness of treatment and diagnostic manipulations and various medical technologies. Conclusion. It is necessary to conduct an expert assessment of regulatory and methodological documents in the field of epidemiological surveillance and sanitary and hygienic measures for the prevention of medical aid related infections of parasitic origin, to optimize the regulatory and methodological base, to develop a number of preventive measures aimed at stopping the spread of parasitic infections in the medical network.

Introduction. The main and the most significant issues of the management of disinfection measures taking into account modern trends and factors affecting the quality of these measures in relation to parasitic infections in the territory of the Russian Federation are considered. The professional community of physicians, epidemiologists, hygienists, disinfectants was almost powerless in the fight against slow or latent infections of a parasitic nature, which are still ca lled “neglected diseases” (“unnoticed diseases”), partly because they occur without pronounced symptoms, and there are a number of unresolved problems in the prevention and diagnosis. Despite the fact that modern medicine in the field of prevention and diagnosis of some infectious diseases has achieved good results, the problems with the disinfection of parasitic pathogens remain relevant. Environmental degradation, various natural disasters, urbanization, increased anthropogenic pressure, as well as climate change do not contribute to improving the difficult situation with regard to the spread, reproduction and formation of new stable foci of parasites. The measures for the improving the system of non-specific prevention of parasitic diseases are determined. Material and methods. In the course of the work, there were analyzed statistical reports of the Federal Reserve and the literature data. Results. The analysis of statistical data showed that during 2014-2018, the incidence of parasites in the Russian population remained at a high level and did not have a pronounced tendency to decrease, and the proposed anti-parasitic disinfectants cannot meet modern anti-epidemiological and hygienic requirements. Conclusion. A number of recommendations and initiatives on disinfection measures for the prevention and spread of parasitic infections in the territory of the Russian Federation were proposed to solve the tasks. There are determined main ways of improving the system of nonspecific prevention of parasitic diseases.

Introduction. The proposed criteria for the sanitary-bacteriological assessment of the quality of tap water must ensure its epidemic safety. In conditions of intensive bacterial contamination of water bodies, a special role is played by the barrier function of water treatment plants in relation to infectious agents. The overall quality of microorganisms is the pronounced resistance in the aquatic environment, primarily resistance to a number of chlorine-containing disinfectants, which guarantee the preservation of the population in drinking water undergone a water treatment system. Therefore, it is necessary to consider other possible ways of disinfection, such as ultraviolet irradiation. Determination of the effective dose of ultraviolet (UV) disinfection against bacterial, viral and parasitic contamination of drinking water. Material and methods. Tap water was used as model water for research. The effectiveness of UV irradiation with doses of 25, 40, 60 mJ / cm2 against microorganisms was studied. Results. In the course of the work, it was established that the UV disinfection technology with a dose of at least 25 mJ/cm2 can be recommended as a disinfection method when used in conjunction with chlorination. Conclusion. In water treatment technology, a dose of at least 25 mJ/cm2 of UV irradiation can be recommended as a method of disinfecting water in case of microbial contamination by bacteria and viruses at a concentration not exceeding nˑ102 cells/virions in 100 ml, and at a concentration of microbiological contamination nˑ103 cells/virions in 100 ml of water, the use of UV disinfection can be recommended only in conjunction with chlorination and with the provision of indices on the residual chlorine in the distribution network before serving to the consumer. The presented scheme will increase the barrier role of water treatment facilities with respect to viral and bacterial contamination, provide a prolonged decontamination effect, contributing to the suppression of bacterial growth in breeding nets and limit the level of parasitic water contamination during water treatment.

Large series of patients with acute myelogenous leukemia (AML) after ex vivo T cell-depleted (TCD) allogeneic hematopoietic stem cell transplantation (allo-HSCT) have not been reported previously. We retrospectively analyzed the outcomes of 266 patients (median age, 54 years) with AML who received CD34-selected TCD allo-HSCTs while in first (75%) or second (25%) complete remission (CR1/CR2) at a single institution. The conditioning regimens were all myeloablative, and no additional graft-versus-host disease (GVHD) prophylaxis was given. The cumulative incidences of grade II-IV and grade III-IV acute GVHD at 180 days were 14% (95% confidence interval [CI], 10% to 18%) and 3% (95% CI, 1% to 5%), respectively. The cumulative incidence of chronic GVHD at 3 years was 3% (95% CI, 1% to 6%). The 3-year cumulative incidence of nonrelapse mortality was 21% (95% CI, 16% to 26%) and that of relapse was 21% (95% CI, 17% to 27%). Overall survival (OS) and disease-free survival (DFS) at 1, 3, and 5 years were 75%, 61%, and 56% and 68%, 57%, and 53%, respectively. There were no significant differences in OS, DFS, and relapse rates for patients who underwent transplantation in CR1 and those who did so in CR2. However, patients with high-risk cytogenetics at diagnosis had significantly poorer outcomes. The OS and DFS rates compare favorably with those for unmodified allo-HSCT, but with considerably lower rates of GVHD.

Background: Diagnosis and remission status at the time of allogeneic haematopoietic stem-cell transplantation (HSCT) are the principal determinants of overall survival following transplantation. We sought to develop a contemporary disease-risk stratification system (DRSS) that accounts for heterogeneous transplantation indications. Methods: In this retrospective cohort study we included 55 histology and remission status combinations across haematological malignancies, including acute leukaemia, lymphoma, multiple myeloma, and myeloproliferative and myelodysplastic disorders. A total of 47 265 adult patients (aged ≥18 years) who received an allogeneic HSCT between Jan 1, 2012, and Dec 31, 2016, and were reported to the European Society for Blood and Marrow Transplantation registry were included. We divided EBMT patients into derivation (n=25 534), tuning (n=18 365), and geographical validation (n=3366) cohorts. Disease combinations were ranked in a multivariable Cox regression for overall survival in the derivation cohort, cutoff for risk groups were evaluated for the tuning cohort, and the selected system was tested on the geographical validation cohort. An independent single-centre US cohort of 660 patients transplanted between Jan 1, 2010, and Dec 31, 2015 was used to externally validate the results. Findings: The DRSS model stratified patients in the derivation cohort (median follow-up was 2·1 years [IQR 1·0–3·2]) into five risk groups with increasing mortality risk: low risk (reference group), intermediate-1 (hazard ratio for overall survival 1·26 [95% CI 1·17–1·36], p