Vol 8, No 4 (2018)
- Year: 2018
- Published: 30.12.2018
- Articles: 295
- URL: https://iimmun.ru/iimm/issue/view/35
- DOI: https://doi.org/10.15789/2220-7619-2018-4
Full Issue
MOLECULAR BASES OF EPIDEMIOLOGY, DIAGNOSTICS, PREVENTION AND TREATMENT OF INFECTIOUS DISEASES
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JUBILEE 2018: 110th ANNIVERSARY OF THE FOUNDATION OF THE ST. PETERSBURG PASTEUR RESEARCH INSTITUTE OF EPIDEMIOLOGY AND MICROBIOLOGY AND THE 95th ANNIVERSARY OF THE NAMING OF THE INSTITUTE AFTER PASTEUR
Abstract
Abstract. The article presents not previously published archive data and copies of documents about Lieberman Yakov Yulievich, the first director of the Petrograd Bacteriological Institute named after Pasteur (now called Saint Petersburg Pasteur Institute). Based on the personal testimony of Ya.Yu. Lieberman, Pyotr Petrovich Maslakovets and Georgii Dmitrievich Belonovsky (the founders and leaders of the Institute) preserved in the archive funds of the Directorate of the Federal Security Service (FSB) of Russian Federation for St. Petersburg and the Leningrad Region, and taking into account the Act of the Commission for the Institute Inspection from December 1930, provided in the article, the authors consistently trace the path of transformations that the Institute has undergone since its foundation. These data are illustrated by copies of publication advertisements contained in the annual city reference book "All Petersburg" for years 1910, 1911, 1912, 1914, 1923, 1924, and 1925 (from the funds of the National Library of Russia). The article also contains copies of official documents dated 1913 and 1914 and stored in the funds of the Central State Historical Archive of St. Petersburg. All documents provided establish the fact that the date of creation of the Institute is 1908.
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DISCIPLES OF PASTEUR FROM RUSSIA
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Abstract. Outstanding scientist, microbiologist, chemist Louis Pasteur left a great legacy to future generations: many discoveries in the relevant fields of science, a large number of disciples, followers and the Paris Institute, bearing his name. Among Pasteur's disciples were more than a hundred Russian scientists, whose fate was different. Some of them returned Home and were able to make a significant contribution to the development of microbiology and immunology in Russia. Not all the names of Pasteur's disciples are known to a wide range of microbiologists. Therefore, the article presents brief materials about some more or less well known scientists-followers of Louis Pasteur. L.S. Tsenkovsky may be considered the founder of microbiology in Russia, who in 1882 published the book "Microorganisms", and later was able to independently create a vaccine against anthrax. Numerous trips to the Pasteur Institute in Paris had a positive value — Lev Semenovich was able to assess how to be equipped laboratories that deal with serious problems in the field of microbiology. Students of Pasteur L.L. Heydenreich, A.D. Pavlovsky, M.F. Popov, A.I. Sudakov, A.A. Raevsky was able not only to develop scientific areas in microbiology, but also to transfer their knowledge to students of medical and veterinary faculties of universities in various cities of Russia, students of the St. Petersburg Military medical Academy and military doctors. I.I. Mechnikov and N.F. Gamalea were great friends and colleagues of Pasteur for many years. Together with Pasteur, they made a great contribution to the prosperity of the Paris Institute and the maintenance of the glory of its founder. The role of these scientists in the world heritage in the field of immunology and microbiology is difficult to overestimate. Associate I.I. Mechnikova was also D.K. Zabolotny, who managed to organize the departments of microbiology and epidemiology in several higher educational institutions of Russia and Ukraine. And, of course, the unsurpassed contribution to the organization of the Pasteur Institute in Russia on the model of the Paris Institute belongs to G.D. Belonovsky. Thanks to his efforts, the Sero-diagnostic laboratory gradually developed into the St. Petersburg Pasteur Institute. Many disciples of Pasteur laid the foundation of a number of scientific areas in Russia: microbiology, immunology, toxicology, hygiene. Thanks to the efforts of enthusiasts among the followers of Louis Pasteur managed to establish in Russia (St. Petersburg) Institute, similar to Paris, also named after him. Each of the disciples of Pasteur thanks to selfless work was able to prove his high rank and make a significant contribution to the development of science for the health of the population of the Homeland.
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PYRAZINAMIDE/PYRAZINOIC ACID RESISTANCE IN MYCOBACTERIUM TUBERCULOSIS: RECENT FINDINGS AND IMPLICATIONS FOR IMPROVING THE TREATMENT OF TUBERCULOSIS
Abstract
Abstract. Pyrazinamide (PZA) is unique in that it is a component of the first line therapy for drug sensitive tuberculosis and in most current and experimental treatments also for multi drug resistant tuberculosis. Furthermore, PZA has been shown to help to ensure lasting cure and prevent relapse in shorter multi drug regimens. PZA is a prodrug. Mycobacterial tuberculosis (MTB) PncA enzyme activates the anti-mycobacterial prodrug PZA by transforming it into pyrazinoic acid (POA). The majority of clinical PZA resistant isolates contain mutations within the pncA gene and therefore remain sensitive to POA as they no longer activate PZA. Resistance to the active compound POA requires an alternative resistance mechanism and in vitro selected spontaneous MTB POA resistant mutants typically acquire a range of mutations in panD or mutations in one of a series of genes most of which are associated with the regulation of the bacterial stringent response. Clinically isolated PZA resistant MTB strains resistant to PZA and POA with mutations in any of these genes are unusual. Thus, it is likely the stringent response is critical for MTB in vivo and a damaged stringent response results in at least a reduction in fitness. Various lead compounds that disrupt the MTB stringent response have been identified that might form the basis for drugs with activity against latent mycobacteria with the potential to shorten tuberculosis treatment. Here we discuss the role of latency in the lifecycle of MTB and possible links to the activity PZA with a focus on potential new targets and drugs.
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BIOFILM FORMATION INDUCED BY CLINICAL ISOLATES OF MYCOBACTERIUM TUBERCULOSIS
Abstract
Abstract. The data proving low probability of observing Biofilm Formation (BF) by contemporary clinical strains of M. tuberculosis growing on liquid medium in vitro are discussed. A hypothesis about the role of MDR/XDR development hindering BF production was proposed. It was found that strains capable of producing BF grow on Lewenstein–Jensen medium generated R-form specific colonies shaped as a disk with a convex center, “UFO-colonies”. Sixty seven “UFO”- strains were investigated to BF production, resistance to antibiotics and their belonging to the main epidemics clusters of the Beijing genotype (CC1 and CC2-W148). It was shown that MDR/XDR strains were also capable of BF production that, however, was remarkably more frequent in strains of CC1 and CC2-W148 genotypes. Thus, it was hypothesized that BF production might potentially influence an outcome of chronic forms of TB-infection.
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GENETIC DIVERSITY OF MYCOBACTERIUM AVIUM subsp. HOMINISSUIS STRAINS ISOLATED IN ITALY BASED ON VNTR LOCI ANALYSIS
Abstract
Abstract. Background. Mycobacterium avium subsp. hominissuis (MAH) is an important pathogen responsible for most of the human-associated nontuberculous mycobacteria infections. Over the past few decades the incidence of MAH infections is increasing in Italy, as in many countries worldwide. The present study is aimed to elucidate the genetic characteristics of MAH strains isolated from human patients using VNTR typing and to show the genetic relatedness among them.
Methods. The genetic diversity of 108 human isolates of MAH was determined by VNTR analysis targeting 8 loci, coded 32, 292, X3, 25, 3, 7, 10 and 47. Results. The VNTR analysis revealed 25 distinct VNTR patterns; of these, 13 patterns were unique, while 12 patterns were shared by 2 or more isolates, thus yielding 12 clusters including a total of 95 isolates. The discriminatory power of our VNTR analysis yielded an HGDI of 0.990, indicating that VNTR typing has an excellent discriminatory power. No association of a particular VNTR pattern with a particular clinical feature, such as the disseminated, pulmonary or extrapulmonary type of infection, was observed. Minimum spanning tree analysis showed that 21 VNTR patterns, occurring either as clustered or unique isolates, differed from the nearest one for one allelic variation.
Conclusions. The results obtained through the VNTR analysis showed that most MAH strains displayed a close genetic relationship. This high phylogenetic proximity of the VNTR loci over a long time period supports the concept that the MAH genotype is highly homogeneous in our geographical area, suggesting the hypothesis of the presence of possible sources of infection and transmission pathways at the local level.
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RETROSPECTIVE ANALYSIS OF SLOVENIAN MYCOBACTERIUM AVIUM COMPLEX AND MYCOBACTERIUM ABSCESSUS COMPLEX ISOLATES AND MOLECULAR RESISTANCE PROFILE
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Abstract. Mycobacteria belonging to Mycobacterium (M.) avium complex (MAC) and M. abscessus complex (MABSC) are the most frequent causes of mycobacteriosis in the world. In the last few years MAC and MABSC taxonomy was rapidly changing due to new molecular methods conveying the possibility to differentiate between species. New techniques are able to identify M. chimaera that was previously recognized as M. intracellulare and also differentiate subspecies of MABSC. Due to their natural habitat, non-tuberculous mycobacteria (NTM) are constantly exposed to various concentrations of antimicrobial drugs and other chemicals and consequently they had developed different mechanisms of resistance. Macrolides and aminoglycosides are frequently used drugs to treat MAC and MABSC infections. The aim of our nation-wide survey was to obtain information about MABSC subspecies prevalence in Slovenia and to assess the percentage of misidentifications of M. chimaera isolates as M. intracellulare in the past. Moreover, the purpose of our study was to reveal, which of the two species M. intracellulare or M. chimaera is clinically more relevant in Slovenia. Further, the aim of the study was to detect mutations in erm(41), rrl and rrs genes, which are known to convey macrolide resistance (erm(41) and rrl) and aminoglycoside resistance (rrs). One hundred and thirty-two Slovenian mycobacterial isolates obtained from the National Mycobacterial Collection that belong to MAC and MABSC were analysed. GenoType NTM-DR was used to differentiate M. intracellulare from M. chimaera and subspecies of MABSC. Our results showed that 48% of previously identified M. intracellulare isolates were actually M. chimaera isolates and that M. abscessus subsp. abscessus was the most frequent subspecies of MABSC. Most of the MABSC isolates carried the inducible macrolide resistance genes (erm(41) and rrl), however none of the isolates of MAC and MABSC had mutations in rrs genes for aminoglycoside resistance.
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CURRENT POSSIBILITIES AND POTENTIAL DEVELOPMENT OF MOLECULAR ENTEROVIRUS SURVEILLANCE. EXPERIENCE OF RUSSIAN FEDERATION
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Abstract. Enteroviruses are small RNA viruses, which are ubiquitous and commonly cause outbreaks with various clinical manifestations. In 2006, the Program on enterovirus surveillance was approved in the Russian Federation. Over the last years, molecular-biological and bioinformatics methods for enterovirus epidemiology studies have been developed both in Russia and worldwide. Currently, identification of enteroviruses is carried out by analyzing nucleotide sequence of the full-length VP1 genome region (ca. 900 nt). Routinely, it is sufficient to obtain a partial VP1 genome region sequence (ca. 300 bp) for enteroviruse verification in most cases; however, a more stringent type criterion of 80% sequence identity should be used compared to the 75% sequence identity cut-off for the complete VP1 genome region. Further sequence analysis may be performed by using Bayesian phylogenetic methods, which allow using molecular clock to trace outbreak emergence. Enteroviruses accumulate about 0.5–1% nucleotide substitutions per year. Therefore, a short genome fragment may be used to analyze virus phylodynamics at the level of international transfers and circulating virus variants. On a shorter timescale, a full-length VP1 genome region or a complete genome sequence are preferred for investigating molecular epidemiology, because a short sequence allows to reliably distinguish not more than 1–2 transmission events per year. Thus, determining enterovirus sequences for full-length VP1 genome region or full-genome sequence is preferred for examining viral outbreaks. It is increasingly apparent that analyzing available enterovirus nucleotide sequences reveals limitations related to uneven surveillance efficacy in various countries and short length of genome fragment measured in routine control. As a result, a proper global-scale analysis of enterovirus molecular epidemiology remains problematic. Over the last 20 years, the number of available enterovirus nucleotide sequences increased by hundred times, but understanding emergence of enterovirus infection outbreaks remains limited. Further development of enterovirus surveillance would require new methods for sewage monitoring, affordable high-throughput sequencing and harmonization of global surveillance systems.
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IMPLEMENTATION OF THE WORLD HEALTH ORGANIZATION WESTERN PACIFIC REGIONAL PLAN OF ACTION FOR MEASLES ELIMINATION
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Abstract. The Western Pacific Region (WPR) is comprised of 37 countries such as China, Japan, Mongolia, Republic of Korea, The Socialist Republic of Vietnam, Papua-New Guinea, Australia, including Pacific Island Countries and Territories (21 countries of PICTs, approx. 3 million people) etc., with a population of 1.85 billion people. Among them, China is the largest and most populous (1.3 billion people) country of the Region. Large measles outbreaks were documented to occur in the Region. In 2003, the Regional Committee announced officially about the WPR action plan on measles elimination 2005, which, however, failed. Since 2012, WPR countries joined the WHO 2012–2020 Global Measles and Rubella Strategic Plan performing a routine measles vaccination (national immunization schedule) or within Expanded Programme on Immunization (EPI). Basically, a two-dose immunization strategy is followed in the WPR countries. Since 2002, measles supplementary immunization activities (SIAs) in children were conducted in the following countries: Japan, Laos, Vietnam, Philippines, Mongolia, Cambodia, Papua New Guinea, and China. Starting from 2005, measles management was considerably improved, demonstrating by 2012 decreased measles incidence rate down to 5.9 cases per million population. In last years, a decreased measles immunization coverage in decreed population groups was noted in the WPR countries that resulted in 2013–2015 measles epidemic involving almost all regional countries. In particular, in China measles incidence rate was 19.6 cases per million population, whereas in the Vietnam Papua New Guinea and Philippines it progressively increased reaching 182.8, 345.9 and 548.0 cases per million population, respectively. Early children not vaccinated according to schedule, adolescents and young adults dominated among measles patients. It was found that measles outbreaks were due to missed vaccination and increased level of vulnerability to measles. Children under one, adolescents and young adults who did not receive a two-dose measles vaccination were in risk group. Analyzing WPR measles epidemiology demonstrated that refusal of parents to vaccinate children, poor knowledge of advantages related to vaccination, insufficient immunization coverage in immigrants, travelers, subjects changing place of residence, workers of healthcare and educational facilities require special attention. In 2017–2018 season, the following measles genotypes were found in the WPR: D8 — Australia, New Zealand, Republic of Korea, Singapore, Japan; Н1 — China; В3 — Philippines, Australia and Japan; D9 — Singapore, Australia, Macau (China), Malaysia and Japan, Н2 strains endemic in Vietnam. According to the WHO, measles endemic transmission has been successfully interrupted; Australia, Macau, Mongolia and Republic of Korea are being verified to eliminate measles; Hong Kong (China) and Singapore (based on available information) are ready to verify measles elimination. Thus, in the Western Pacific Region measles elimination is achievable after solving current issues such as increasing and maintaining high-level routine vaccination and conducting measles supplementary immunization campaigns in epidemically important contact clusters.
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SUMMARY OF INFLUENZA AND OTHER RESPIRATORY VIRUSES DETECTED AND CHARACTERIZED IN RUSSIA DURING 2017–2018 SEASON
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Abstract. The influenza season 2017–2018 started significantly later compared to the five previous seasons. Influenza epidemic lasted for 12 weeks (weeks 6–17), was of moderate intensity and 10,4% of the population of the country was involved with children aged 0–2 and 3–6 years being the most affected groups as usually. The average hospitalization rate of patients with ILI and ARI was 2,6% and was the highest in infants aged 0–2 years (5,4%). The number of influenzaassociated deaths was two times higher this season compared to 2016–2017 which can be attributed to the circulation of A(H1N1)pdm09 viruses that still is the major cause of lethal influenza outcomes in the country. A total 72 759 patients were investigated by RT-PCR in 55 collaborating RBLs. Laboratory confirmed influenza (LCI) was detected in 12 149 (20.7%) cases, of which 39.3% were influenza A(H1N1)pdm09 viruses, 29.6% were A(H3N2) and 31.1% influenza B (Yamagata lineage) viruses. The first cases of influenza viruses were detected at the very beginning of the season (weeks 40–45.2017), however a distinct increase in the rate of detection was registered only from the week 2.2018 with the peak on the week 13–14.2018 and subsequent gradual decline up to the end of the season. The certain differences in the etiology of morbidity between Federal Districts were registered. The impact of influenza and other ARI agents in different stage of epidemic was determined. In the pre-epidemic period, the incidence growth was occurred mainly due to ARI agents (about 32,7%), especially due to rhinoviruses (RhV) and RSV (10.2 and 8.0% cases, respectively) while LCI were registered in 3.4% only. During the epidemic, the rate of LCI detection increased up to 29.2% at simultaneous decrease in frequency of parainfluenza, adenovirus, bocavirus, coronavirus and, especially, rhinoviruses, to a lesser extent RSV infection. In the post-epidemic period, the role of influenza A(H1N1)pdm09, A(H3N2) and В viruses decreased up to 6.1; 6.9 and 3.6%, respectively, with increase of rhinoviruses (9.5% of diseases). Genetic analysis of influenza A(H1N1)pdm09 and A(H3N2) viruses circulating in 2017–2018 season showed that all analyzed viruses by the structure of surface genes encoding antigenic determinants, in difference from influenza B viruses, corresponded to the vaccine strains recommended by WHO for the Northern Hemisphere for 2017–2018 epidemic season. However, significant changes in the internal genes of circulating viruses were revealed. The control of the susceptibility of 316 influenza A and B viruses to antiviral drugs showed that the absolute majority of them (99.7%) retained their susceptibility to neuraminidase inhibitors.
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NEW OPPORTUNITIES TO IDENTIFY AND TYPE STAPHYLOCOCCUS spp. BY USING MALDI-TOF MASS SPECTROMETRY
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Abstract. Mass spectrometry profiles of microorganisms obtained by time-of-flight matrix-associated laser desorption/ionization (MALDI-TOF) mass spectrometry are a source of information about peptide profiles can be used for microbial identification and typing. A variety of technical and bioinformational solutions complicate developing of a united mass-spectro-profile database. Staphylococcus spp. strains are good studied objects for identification by MALDI-TOF mass spectrometry, frequently resulting in nosocomial infections, especially in immunocompromised patients. Rapid differentiation of nosocomial, multiresistant and highly virulent isolates of Staphylococcus spp. Allows to reduce the lethality in weakened and immunocompromised patients. The study was aimed at assessing comparability and reproducibility of identification and typing results for Staphylococcus spp by MALDI-TOF mass spectrometry. Comparing 292 Staphylococcus spp. isolates in clinical specimens obtained fron the multidisciplinary hospital at the NWSMU im. I.I. Mechnikov was carried out by using MALDI-TOF mass spectrometer BactoSCREEN ID (Litech, Russia) and Bruker Biotyper 3.1 (Bruker GmbH, Germany). Comparability of Staphylococcus spp. identification showed that 95.9%; 12 isolates (4.1%) by “Bruker Biotyper 3.1” and 3 isolates (1.1%) by using “BactoSCREEN ID” were incorrectly identified. Repeated identification leveled the differences between the systems used. In addition, it was shown that the method of protein extraction did not affect reliability of Staphylococcus spp. species identification by using databases (÷2, p > 0.05) compared to intraspecific typing (÷2, p < 0.0001). Using different extraction protocols showed that Staphylococcus spp. mass-spectra differed by peak intensity level within the mass range up to 4000 m/z, 5300±600 m/z and 6500±500 m/z, as well as higher than 7000 m/z. Peaks of low-molecular weight peptides were detected under full protein extraction compared to sample preparation on plate extraction. To develop a unified protocol for mass-spectrometry profile processing, a reliability of the basic statistical variables (mode, median, maximum, minimum and arithmetic mean) was evaluated. Analysis of the median mass spectrometry profiles is recommended for Staphylococcus spp. intraspecific typing by using MALDI-TOF mass spectrometry as the most reproducible and consistent approach. As a result, two systems for MALDI-TOF mass spectrometry reliably identify Staphylococcus spp., but standardization of sample preparation and bioinformation analysis is required for Staphylococcus spp. typing.
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TIME-KILL ASSAY: AN EFFICACY OF SYNERGY BETWEEN CARBAPENEMS AND CLODRONIC ACID
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Abstract. Currently, a search for augmenting antibiotics activity is still crucial due to elevated frequency of detecting carbapenem-resistant Gran-positive bacterial isolates. To resolve this, it might be reasonable to combine carbapenems metal-â-lactamase (MâL) inhibitors. Unfortunately, no MâL inhibitors approved for treatment of carbapenem-resistant infections are currently available. Pathogenic bacteria may survive antibiotic attack, exert tolerance and persistence accompanied with the ongoing infectious process. In connection with this, determining dependence between antimicrobialrelated bactericidal effect and exposure time on microbes at 4, 8, 12 and 24 hours after the onset, a so called time-kill assay, is necessary. A synergy between both agents was noted upon reduced microbial population by ≥ 3 log10. A checkerboard array followed by seeding the microplate well contents onto a dense nutrient medium at various time points were used to assess a synergistic efficacy of carbapenems applied together with clodronic acid against MâL-producing VIMgenotype P. aeruginosa 532/14 clinical isolate obtained from patients with infectious complications (minimal inhibitory concentrations [MIC] for imipenem or meropenem were 512 μg/ml), microbial burden 106 CFU/ml. Optical density was measured at two wavelengths (490 and 630 nm) in ELx800 reader, within 4–24 hour exposure time to determine time of logarithmic growth phase emerging in test culture. It is noteworthy that magnitude of optical density is a difference between two bichromatic measurements resulting in remarkably reduced inaccuracy due to scratches or fingerprints left on the plate. It was found that clodronic acid exhibited a synergic bactericidal effect with carbapenems against a clinically resistant MâL-producing VIM-genotype P. aeruginosa 532/14 strain. Upon that, imipenem-related antimicrobial activity was evident as early as 8 hours after the onset decreasing cell growth down to 1.4 log10 compared to control, whereas 12 hours later it resulted in total inhibition of test strain by decreasing growth of the test strain by 6 log10. Meropenem in combination with clodronic acid showed a more pronounced activity: complete absence of P. aeruginosa 532/14 growth by 8 hours of incubation, growth suppression by 3.2 log10, which reached 6 log10 12–24 hours after the onset. Time-kill assay allows to identify efficient combinations of carbapenems and MâL inhibitors, which is of great importance for increasing therapeutic efficacy of patients with severe purulent-septic complications.
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RESULTS AND DIRECTIONS OF ACTIVITIES TO ENSURE THE EPIDEMIOLOGICAL SAFETY OF THE POPULATION IN MODERN CONDITIONS
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THE PREVALENCE AND MOLECULAR EPIDEMIOLOGY OF STENOTROPHOMONAS MALTOPHILA IN THE INTENSIVE CARE UNITS
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MODERN METHODS OF MOLECULAR DIAGNOSTICS OF INFECTIOUS DISEASES
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DETECTION AND ANALYSIS OF CRISPR-CAS SYSTEMS IN PLASMIDS OF DIFFERENT BACILLUS THURINGIENSIS STRAINS
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VIRAL INFECTIONS MANAGED BY MEANS OF VACCINATION AT THE STAGE OF DESTRUCTION AND ELIMINATION
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ZOONOTIC AND PARASITIC INFECTIONS: CLINICAL, EPIDEMIOLOGICAL AND LABORATORY ASPECTS
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YERSINIOSIS: TAXONOMY, PHYLOGEOGRAPHY, POLYMORPHISM OF PATHOGENICITY FACTORS AND SELECTIVE VIRULENCE
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MONITORING OF VACCINAL PROCESS IN HUMANS RESIDING IN THE ALTAI HIGH-MOUNTAINOUS NATURAL PLAGUE FOCUS
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TUBERCULOSIS AND MYCOBACTERIA: MOLECULAR APPROACH
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HIV, HEPATITIS AND OTHER SOCIALLY SIGNIFICANT INFECTIONS
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INFECTIOUS IMMUNOLOGY AT THE PRESENT STAGE
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INNATE AND ADAPTIVE IMMUNITY CYTOKINES IN NASAL MUCOSA AND BLOOD SERUM OF ALLERGIC RHINITIS PATIENTS
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ANTIBIOTIC RESISTANCE OF MICROORGANISMS: CURRENT ISSUES OF DIAGNOSIS AND WAYS TO OVERCOME
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NEW CHEMOTHERAPY FOR THE TREATMENT OF INFECTIOUS DISEASES
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EDITORIAL
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