Nature of the epidemic process in Sonne and Flexner dysenteries and the causes of its induction

A comparative study of the epidemic process in Sh. sonnei and Sh. flexneri dysentery in different regions of the USSR revealed that the morbidity level of Sh. sonnei dysentery changed simultaneously in the regions under study at intervals of 2-3 years. Sh. flexneri dysentery showed morbidity rises occurring at intervals 6-8 years, and their occurrence did not coincide with the periods of elevated morbidity in Sh. sonnei dysentery. The data obtained in the cohort analysis and in the study of recurrent morbidity suggest that Sh. flexneri dysentery produces more pronounced postinfection immunity than Sh. sonnei dysentery, and the immunological factor probably affects the dynamics of the epidemic of these Shigella infections.     

Characteristics and trends in the development of an epidemic process of Sonne and Flexner dysentery among preschool children

The results of studies indicate that the morbidity rates of dysentery among children attending preschool institutions and children brought up at home converged in recent years. This phenomenon was most pronounced among children of the kindergarten age group. At the same time dysentery caused by Sh. sonnei and Sh. flexnery produced a higher morbidity rate among children attending nursery in comparison with that among children of the same age group brought up at home. Group infections in preschool institutions were caused by Sh. sonnei in 89.1% of cases and by Sh. flexneri in 10.9% of cases. Outbreaks due to the transfer of infection through everyday contacts were observed only in dysentery caused by Sh. sonnei, constituting 71.4% of the total number of dysentery outbreaks.     

Manifestations of the cyclic nature of the epidemic process in dysentery

Different forms of dysentery, especially those caused by Sh. sonnei and Sh. flexneri, have been found to differ considerably in their cyclic recurrence. The development cycles of the epidemic processes of dysentery have an objective character, occur in the presence of any tendencies in the morbidity rate, and depend on the natural factors. Thus, the cycles of increase and decrease in morbidity are 3, 6, 9, 12 years for dysentery caused by Sh. sonnei and 6, 7, 8 years for dysentery caused by Sh. flexneri.     

Use of data on Shigella contamination and immunity indices for studying the activity of the epidemic process in dysentery

The comparative study of the monthly distribution of characteristics indicating the levels of contamination with Sh. sonnei and Sh. flexneri separately, as well as the seasonal dynamics of the corresponding antibodies, in the years with high and low morbidity levels has been made with the use the indirect hemagglutination test. The possibility of using these characteristics for the evaluation of the activity of the epidemic process in dysentery caused by Sh. sonnei is shown.     

Data to substantiate the early immunological diagnosis of dysentery]

The interaction of the whole blood from patients with dysentery and gastrointestinal diseases of non-dysenteric etiology, with the causative agents of dysentery, Sh. sonnei and Sh. flexneri, and saprophytic, staphylococci labeled with radioactive isotopes was studied in vitro. In dysentery an increase in the capacity of the blood for Shigella fixation was observed from the beginning of the disease. During the 1st week of the disease this reaction was strictly specific and accompanied by a decrease in the fixation of staphylococci, but later the reaction became relatively specific. An increase in Shigella fixation occurred considerably earlier than immune antibody formation, as revealed by the indirect hemagglutination test. This research substantiates the possibility of an earlier immunological diagnosis of dysentery as compared with the serological methods.     

Use of intraspecific differentiation of Sh. sonnei for the purpose of studying patterns in the epidemic process of dysentery]

The use of complex typing of Sh. sonnei by the colicinogenic and enzymatic properties in epidemiological analysis of dysentery caused by Sh. sonnei, apart from establishment of epidemiological connections in the foci, offered a possibility of introducing significant corrections in the determination of the sizes and duration of existence of the epidemic foci, as well as in detection of the sources and factors of transmission of the infection established in epidemiological examination. Typing of Sh. sonnei also aided in ascertaining that, apart from general regularities in the individual territories, the epidemic process in individual groups of the population and in different seasons of the year could also course autonomously.     

Tendencies in the epidemic process of dysentery in Ryazan

In shigellosis caused by Sh. sonnei and Sh. flexneri the epidemic process was found to have considerable difference in the tendencies and pace of its development. Shigellosis which dominated in the etiological structure at the period following 1966 was dysentery caused by Sh. sonnei; it showed the tendency towards a decrease in morbidity rate. Dysentery caused by Sh. flexneri was second in respect to the frequency of its occurrence after 1966 and showed the tendency towards increase. The simultaneous circulation of Sh. sonnei and Sh. flexneri, the differences in the epidemiology of these types of shigellosis make it imperative that they be studied separately, taking into ccount their etiological selectivity to the main routes of transmitting the infection.     

Modern methods of intraspecies typing of Sonne shigellae. II. Spread of Sonne shigellae of different biochemical types]

The paper presents the results of studying peculiarities of the Sh. sonnei of different biochemical types spread established by their typing scheme suggested by the authors earlier according to rhamnose, xylose and maltose. The epidemic process in dysentery both during the years of the rise and of the decline of its incidence at various territories of the countries proved to be maintained on account of circulation of Sh. sonnei of various biochemical types. The results of studying their dissociative and virulent properties confirmed the biological separation of individual biochemical types. An interrelationship between the character of the biochemical pattern of Shigellae sonnei at the individual territories and the persisting activity of different ways of dysentery transmission was determined. The results of studying the biochemical pattern could be used as an indicator of the degree of activity of individual ways of dysentery spread at various territories.     

Determination of the O antigen of Shigella sonnei by a competitive immunoenzyme method

A technique for immunoenzymatic diagnosis of dysentery by Shigella sonnei O-antigen was developed. For induction of antibodies to O-antigen rabbits were immunized by intravenous administration of a commercial antidysentery vaccine. Specific antibodies to O-antigen belonging to class G immunoglobulins and not binding to O-antigens of Sh. flexneri and Salmonella typhimurium were obtained. beta-Lactamase of Bacillus licheniformis 749/c was used as a marker enzyme in the immunoenzymatic assay. To increase the sensitivity, beta-lactamase molecules were preliminarily linked with glutaric aldehyde into oligomers. Conjugates of Sh. sonnei O-antigen with the oligomers of B. licheniformis 749/c beta-lactamase were prepared with the periodate method by oxidizing O-antigen. The conjugate was used in competing solid phase immunoenzymatic assay for determination of Sh. sonnei O-antigen in blood serum of patients with dysentery. The sensitivity of the assay is 0.5-1 ng per 1 ml of O-antigen.     

Recurrent Sonne dysentery morbidity caused by Shigella of varying enzymatic types]

The proportion of repeated cases of dysentery caused by Sh. sonnei does not exceed 3.59% of the total number of bacteriologically confirmed cases of this infection. The frequency of reinfection depended on the enzymatic type of the causative agent. The rarest cases of reinfection were those occurring after the primary infection with Sh. sonnei, biochemical type 2, whereas the most frequent after the primary infection with Sh. sonnei, type 3. Reinfection with heterologous Sh. sonnei was registered mostly 6-12 months after the primary infection, and with homologous Sh. sonnei after a 1 year and later.     

Latex test used for diagnosis of dysentery caused by Shigella sonnei]

The aim of this study was to evaluate the usefulness of latex reagent coated with immunoglobulins specific for antigens of phase I and II of S. sonnei for detection of these antigens in primary, mixed bacterial cultures. The study was performed on 919 fecal samples from individuals with clinical symptoms of dysentery, convalescents and from contact individuals. Material used for the test was bacterial suspension collected from McConkey or SS agars and a culture from selenite F broth heated at 100C. The results of the latex test were compared with the results of isolation of S. sonnei from the same cultures. S. sonnei was isolated from feces of 140 individuals (15.2%), while the latex test was positive in 215 cases (23.4%). The highest testing effectiveness , significantly higher than when isolation of pathogen was performed, was obtained only when 18-20 hr culture on Selenite F medium was used for latex test. The correlation between efficacy of testing for S. sonnei and phosphate content of Selenite F and a mode of its preparation was found. The latex test allows to eliminate from further bacteriological studies cultures free of S. sonnei thus it gives measurable economical profits and it shortens significantly time period of bacteriological examination.     

Assessment of the diagnostic effectiveness of antigenic preparations from Sonne shigellae]

The authors studied the diagnostic possibilities of 4 diagnistic agents from Sh. sonnei. A total of 1500 from 1122 persons were investigated. Specificity and sensitivity of the diagnostic agents from Sh. sonnei were determined. Methodical priciples of the objective assessment of the diagnostic efficacy of the antigenic preparations in the controlled epidemiological trial were elaborated. A possibility of establishment of the etiological diagnosis of Sonne dysentery in the passive hemagglutination or blast agglutination tests by the results of testing with the diagnostic agents of the serum obtained from the patient once or by determining the dynamics of the antibody titre rise in coupled serum portions was not great.     

Patterns determining the dynamics of the epidemic process in Sonne dysentery and its relation to the type of causative agent]

The results of studying the dynamics of the epidemic process of dysentery, based on the data from 3 districts of Moscow, are presented. The study revealed the periodicity of 3 years in the course of the epidemic process of dysentery, occurring against the background of a considerable predominance of Sh. sonnei, biochemical type 2. The use of the cohort method for analyzing the age structure of dysentery cases showed the possibility of using this method to find out to what extent the epidemic process repeatedly affected the same groups of population.     

Immunoelectrophoretic analysis of the antigenic structure of Sh. sonnei]

The authors studied the antigenic composition of 105 Sh. sonnei strains freshly isolated from patients suffering from acute dysentery and carriers. Immunophoregrams of pure S-and R-forms species were obtained. Up to 13 antigens differing by electrophoretic and diffusion mobility and immunological specificity were revealed among soluble Sh. sonnei antigens The position of common and specific antigens was determined on the immunophoregram. Along with the thermostable somatic O-antigen detected at the I phase of the S-forms, and two thermolabile O-antigen components at the II phase, and the R-forms, there was revealed a surface, relatively thermolabile, K-antigen of A-type capable of agglutinating live bacteria in the O-antiserum; position of the latter on the immunophoregram was also determined.     

Current methods of intraspecies typing of Shigella sonnei. 3. Bacteriophage typing]

Data are presented concerning the study of the phage type of Sh. sonnei isolated in various regions of the USSR. The strains isolated belonged to 64 phage types corresponding to the Hammarstr?m's scheme. A variety of Shigella sonnei phage types both in one and in different territories of the country provided future prospects for successful use of phage typing for the purpose of epidemiological supervision and analysis of dysentery morbidity.     

Experience with the diagnosis of dysentery in adults by determining antibodies to Shigella in the saliva]

Concentration of immunoglobulins and titres of antibodies to Sh. sonnei, Sh. flexneri and enteropathogenic E. coli 0111 was determined in mixed saliva and the blood serum of patients suffering from Sonne dysentery, acute intestinal diseases of unknown etiology, and healthy individuals. The sum total immunoglobulin concentration in mixed saliva proved to be 53--81 times less than in the blood serum, but in the first substrate there was 53--75, and in the second--15% of immunoglobulin A. There proved to be distinct changes in the specific IgA-antibodies in the saliva of patients with Sonne dysentery. A preponderant accumulation of IgG-antibodies was noted in the blood serum. Elevation of both types of antibodies was maximal during the second week of the disease. Sonne dysentery was diagnosed in 80% of the patients by recording the intensity of shifts in the specific antibodies in the saliva, and in 63%--in the blood. The expediency of immunological testing of saliva for the diagnosis of dysentery is substantiated.     

The modulation of hela cells secretory patterns by invasive Shigella spp. and enteroinvasive E. coli bacterial cells and their soluble components

Considering the important role of cytokines in the initiation and evolution of the inflammatory process induced by Shigella and EIEC strains, the purpose of this study was the characterization of the secretory patterns of HeLa cells induced by Shigella ssp. and EIEC strains and to link the obtained results with the invasiveness level of bacterial strains on this cellular line. During this study there were analyzed two EIEC strains and 12 strains of the following Shigella species: 2 Sh. flexneri 2a, 2 Sh. flexneri 3a, 2 Sh. flexneri 4a, 2 Sh. boydii, 2 Sh. sonnei strains isolated in Romania during 2005 from children with dysentery and diarrhoea and confirmed for their invasive ability by Sereny test. The level of the main pro and anti-inflammatory cytokines, IL-1 beta, IL-6, IL-10, IL-13, IL-17 and TNF-alpha induced by whole bacterial cultures as well as by their soluble mediators was determined by an ELISA test. Our results showed that HeLa cells can be used not only for the qualitative and quantitative assessment of Shigella and EIEC strains invasion ability, but also as a simple work procedure for the investigation of an in vitro complex crosstalk communication mechanisms that involves physical interactions between bacterial cells and epithelial cells (adhesins and complementary receptors) and pro- and anti-inflammatory molecules regulation.The majority of the analyzed Shigella serogroups, with the exception of Shigella sonnei and EIEC strains, inhibited the inflammatory response by reducing the expression of majority of pro-inflammatory cytokines, i.e., IL-1 beta, IL-6, TNF-alpha and IL-17. The reduced cost of the in vitro procedure, the possibility of results interpretation and the strict regulations concerning the use of animals for experimental purposes are the main reasons that support the implementation of such an in vitro test in the research labs.     

Study of the methylation process and DNA methylase specificity in Shigella]

The nature and content of minor bases in DNA of 3 Shigella strains are investigated. DNAs from Shigella stutzeri 2, Sh. sonnei 1188 and Sh. sonnei 311 are found to contain 0.43, 0.56 and 0.45 mol.% of N6-methyladenine respectively. 5-methylcytosine (0.16 mol.%) is discovered in Sh. sonnei 311. Substrate specificity of adenine methylase from Sh. sonnei 1188 with respect to phage DNAs of different host modification is investigated. Recognition sites for guanine methylase of DDVI phage and for adenine methylase of Sh. sonnei 1188 turned to be different. DNA of DDII phage grown in Sh. stutzeri 2 cells does not accept methyl groups under the treatment with Sh. sonnei 1188 extracts, but it is methylated by Escherichia coli extract. Adenine methylases of Sh. sonnei 1188 and Sh. stutzeri 2 are suggested to be either the same enzyme, or enzymes, which recognition sites are partially overlapped.     

Development and use of solid-phase test-systems for the detection of O-antigen of Shigella sonnei and Shigella flexneri

The possibility of the diagnosis of dysentery caused by S. sonnei and S. flexneri, as well as the determination of the dynamics of the distribution of specific O-antigen in the patient's body, by means of the enzyme-linked immunosorbent assay system developed on the basis of antibody preparations obtained by immunosorption has been studied. The study has shown that for better diagnosis the use of fecal extracts is preferable in assays; when used in combination with bacteriological analysis, these assays make it possible to increase the confirmation of the diagnosis of dysentery by several fold.     

Differential characteristics of the I and II phases and R-form of Sh. sonnei]

Characteristics of various phases of Sh. sonnei were determined on the standard strains of Sh. sonnei of phases I and II and R-form used for the industrial production of agglutinating monoreceptor sera. Bacteria of phase I displayed a distinct morphological, cultural and serological differences. For the differentiation of Sh. sonnei of the II phase and R-form, representing the greatest difficulty in this respect, it is recommended to use phages RFfm and 6-SR, and also indirect immunofluorescence method with the application of agglutinating monophasic serum against the II phase of Sh. sonnei. In addition, a study was made of over 20 various Sh. sonnei strains at different dissociative process phases. Verification data completely confirmed the specificity of phages RFfm and 6-SR for Sh. sonnei in the II phase. The efficacy of the immunofluorescence method was confirmed on 6 strains.