Comparison of genomes of Coxiella burnetii strains using formal order analysis.

The Coxiella burnetii strain NL3262 was isolated during the Q fever outbreak in the Netherlands in 2007-2010. Formal-order analysis (FOA) was used to study the similarity of the genome (chromosome and plasmid) of this strain with the genomes from other strains. Chromosomes from ten C. burnetii strains and eight plasmids were studied with FOA tools such as 'Map of genes' and 'Matrix of similarity.' The 'Map of genes' tool showed that the chromosome of strain C. burnetii str. NL3262 distanced itself by the index of average remoteness (g) of 1.449640 (x-axis) from chromosomes of other strains (g 1.448295-1.448865). The 'Matrix of similarity' was used for an advanced analysis of the obtained results. The complete similarity of the components of chromosomes and plasmids was determined by pairwise comparison and the identification of nucleotides matching with them. A total of 84.90% of the chromosomal components of C. burnetii strain NL3262 coincided completely with the chromosomal components of strain Z3055. For chromosomes of other strains, this percentage varied from 12.06% to 47.14%. The plasmid of strain NL3262 had 50.0% of the components being completely coincident with the components of the plasmid of RSA 331; with RSA 493 it was 29.89%. Thus, C. burnetii str. NL3262 is the closest to str. Z3055 by the similarity of the chromosomal components, but on the index of average remoteness of the chromosome and the similarity of the plasmids' QpH1 components, it is the closest to strain RSA 331.

Rickettsiologist Pavel F. Zdrodovskii: larger than life, and not just for his famous book.

This article highlights the biography and scientific accomplishments of Pavel F. Zdrodovskii and his contributions to understanding the biology, pathogenesis, treatment, prevention and epidemiology of brucellosis, rickettsioses and many other infectious diseases.

[BRILL-ZINSER DISEASE AS A CONSEQUENCE OF RICKETTSIA PROWAZEKII PERSISTENCE IN PREVIOUSLY ILL WHO HAVE HAD EPIDEMIC TYPHUS (EPIDEMIOLOGIC ASPECTS)].

Materials, that summarize data of original research and scientific literature on epidemiology and problems of persistence during epidemic typhus, whose causative agent (Rickettsia prowazekii) is reactivated in the organism of the previously ill and is manifested as Brill-Zinser disease, are presented. A retrospective analysis was carried out with the data obtained by Russian (All-Union) Centre for Rickettsioses during study of epidemiologic examination maps of 5705 typhus nidi and results of 19 463 blood sera analysis during study of immunologic structure of population in the territories of the former USSR for the period from 1970 to 1992. A decrease of epidemic typhus morbidity and an increase of the fraction of Brill-Zinser disease took place as a result of pediculosis corporis control. In separate territories specific weight of Brill-Zinser disease was 48% in 1952, up to 80% in 1969, and from 1977 all the ill were previously ill. However, during the perestroika period and afterwards, due to a reduction of economic and hygienic living conditions, appearance of refugees, the immune structure regarding typhus began to change. Due to the buildup of the population migration process and the presence of risk groups (refugees, homeless) among population of regions, where local wars are waged, the enhancement of methods of epidemic typhus and Brill-Zinser disease diagnostics and pediculosis corporis eradication is necessary. Study of R. prowazekii by molecular-genetics methods is necessary for complete understanding of its mechanism of persistence.

[Emergence of tick-borne spotted fever group rickettsiosis in Moscow].

AIM: Analysis of clinical cases of tick-borne spotted fever (TSF) group rickettsiosis in 2005 - 2010. MATERIALS AND METHODS: General clinical, biochemical and serological parameters were determined in 10 tick-borne spotted fever group rickettsiosis patients who had visited various geographical regions of the World. RESULTS: TSF group rickettsiosis diagnostic criteria, optimal serological diagnostics timing were determined. Possible diagnostic errors, features of serological diagnostics and antibacterial therapy of this nosologic form are discussed. CONCLUSION: Indication for TSF examination are primarily epidemiologic including tick attachment indication and clinical data. Serological studies are positive only in 3 - 4 weeks after the onset of the infection and thus can not be used for early diagnostics.

[Ecology of Rickettsia and epidemiology of rickettsioses].

Questions of ecology of obligate-transmissible rickettsioses causative agents and their natural environment--organism of blood-sucking arthropoda (ixodes, gamasid ticks, harvest ticks, lices, fleas) are considered. In tick organism Rickettsia are propagated, and transphase and transovarial transmission of rickettsia maintains population in natural focuses. The relationship between Rickettsia ecology and rickettsioses epidemiology is evident. Data about new-revealed and recurrent rickettsioses and role of molecular-biological and genetical methods of Rickettsia detection are represented. Biological and social factors having influence on appearance of new and recurrent rickettsioses are determined with the aim of their prediction.

[Bartonella infection in hematological practice].

AIM: To characterize the clinical and histological features of Bartonella infection in patients asking for hematological advice and to assess the significance of serological and molecular methods for the diagnosis of this infection. MATERIALS AND METHODS: The case histories of 747 patients asking for advice at the Hematology Research Cancer, Russian Academy of Medical Sciences, for lymphadenopaphy were retrospectively studied. The study included 10 patients in whom Bartonella infection could be suspected. For verification of the diagnosis, the authors conducted a serological study of the patients' sera and a molecular study of archival paraffined lymph node biopsy specimens. RESULTS: The study showed it possible to make a retrospective diagnosis of cat-scratch disease (CSD) by polymerase chain reaction (PCR) used in the study of archival lymph node biopsy specimens and stained preparations. CONCLUSION: CSD should be suspected when a patient has sustained lymphadenopathy and a respective epidemiological history (feline contact). Bartonella infection should be diagnosed on the basis of a dynamic serological study and, if possible, PCR of cells from biopsy specimens of lymph nodes or the lesion developed at the site of Bartonella penetration into the human body (primary affect).

[Evaluation of the epidemiological situation in different territories of the Russian Federation, based on the Bank of Standard Blood Sera].

The Bank of Standard Sera created in the Gamaleya Research Institute of Epidemiology and Microbiology provide for evaluation the epidemiological situation in any separate territory and in the whole of the country. The collection of certified blood sera is divided according to different territories of the country and makes it possible to obtain the retrospective and operative information on the level of population immunity and the immune structure of the population with respect to different infective agents, and to reveal the susceptible group of the population. The analysis of the unfavorable character of the epidemiological situation in the inspected territory with due consideration of its climato-geographical and anthropogenic environmental, socio-demographic characteristics, the level of population immunity and the immune structure of the population with respect to different infective agents makes it possible to carry out the epidemiological diagnostic (an outbreak, the import of infection, the use of a bacteriological weapon), prognosis and the prophylaxis of diseases, as well as the epidemiological cartography of territory.

[Epidemiological aspects of the present-day demographic situation in Russia].

Study of reference rate in healthcare institutions in 1991 - 2001 demonstrated a steady growth of general mortality and disease incidence among the population (by 22.7% and 8.1%, respectively). The number of patients with acute and chronic diseases registered in 2001 was by 16.4% higher than that in 1991. These trends were characteristic of all disease classes, but the most intensive growth was observed in such classes as diseases of blood and hemopoetic organs (2.5 times), endocrinal and urogenital diseases (1.8 times). Disease incidence and general mortality among children aged 0 to 14 had grown by 25% and 33.2%, among adolescents--by 60% and 71.1%, among adults--by 5.2% and 20.3%, respectively. These negative trends in population health were accompanied by a constant growth of incapacitation rate in 1990 - 2000 (2.4 times, and 4.3 times among children and adolescents younger than 18). There is a correlation between mortality and traditionally used demographic parameters, which suggests that mortality should be recognized the key demographic parameter. In order to achieve a satisfactory level of population health and adequate demographic transition, and to develop general sanatory strategy, the State should undertake powerful, goal-seeking, society-oriented measures, including deep clinical and epidemiological study of human pathology.

[Bartonella and bartonellosis--emerging and re-emerging infections: epidemiology, clinical course, diagnosis].

Information on the epidemiology, clinic and diagnostics of bartonellosis is updated. The importance of bartonellosis lies in the fact that its main risk group embraces patients with immunodeficiencies of different origin, the number of such patients constantly growing. The diagnostics of bartonellosis is insufficiently developed and the research on Bartonella organisms, except for the trunch (Volynia) fever causative agent, is insufficient in our country.

[Survival of Coxiella burnetii in soil].

For the first time the survival of Coxiella burnetii of five types in soils has been studied. The survival of C. burnetii has been found to depend on the content of organic substances in black earth, as well as soil temperature. The method for the prevention of an epidemic outbreak of Q fever directly under the natural conditions has been proposed.

[Microorganism "Montezuma" of the order Rickettsiales: the potential causative agent of tick-borne disease in the Far East of Russia]. / Mikroorganizm "Montezuma" iz poriadka Rickettsiales: potentsial'nyi vozbuditel' kleshchevogo transmissivnogo zabolevaniia na Dal'nem Vostoke Rossii.

A new microorganism, tentatively named "Montezuma" was detected in ticks and in specimens (blood, bioptic specimens of the primary affect) taken from patients with an acute fever disease, etiologically linked with the bites of Ixodes ticks in the Far East of the Russian Federation. After sequencing the products of the amplification of DNA isolated from ticks with wide-spectrum primers new primers were developed, highly specific to the unusual sequence thus obtained. The study revealed that ticks of the species Ixodes persulcatus (97%) and Haemophysalis concinnae (5%) contained DNA of this microorganism. The same DNA was detected in materials taken from the patients. The phylogenetic analysis of the gene showed that this organism formed an independent and well defined branch within the order Rickettsiales. The nearest homology (89%) was observed with recently detected endosymbiotes Acanthamoeba. The similarity with their relatives from the families Rickettsiaceae and Anaplasmataceae of the order Rickettsiales was within 81-86%, which made it possible to infer the existence of, probably, only a new genus, but also a family. The isolated DNA belonged, supposedly, to the new microoganism which caused a tick-borne disease in humans, transmitted through bites of Ixodes ticks, and was, supposedly, widely spread in the southern area of the Khabarovsk Territory.

[Bartonella and bartonellosis--emerging and re-emerging infections. Taxonomy, bacteriology, pathogenesis and genetics]. / Bartonelly i bartonellezy--novye i vozvrashchaiushchiesia. Taksonomiia, bakteriologiia, patogenez i genetika.

The review updates knowledge on the taxonomy, bacteriology and genetics of Bartonella as well as pathogenesis of bartonellosis. The role of Bartonella in human pathology, formerly considered to be rather modest, causes now growing anxiety. In this connection Bartonella are now believed to be the causative agents of so-called emerging and re-emerging diseases, i.e. diseases, formerly unknown to man, and diseases, believed to be eradicated and playing at present no important role in human pathology. These microorganisms are a bright example of successes of molecular biology in the detection of microorganisms, as well as in their phylogenetics and systematics.

[Genotyping of spotted tick fever rickettsiae, detected in Russia and Kazakhstan]. / Genotipirovanie rikketsii gruppy kleshchevoi piatnistoi likhoradki, vyiavlennykh v Rossii i Kazakhstane.

R. slovaca was first detected in the ticks D. marginatus gathered in the Stavropol Territory and the Voronezh Region (European Russia). The recently discovered rickettsial genotype DnS14 was first found in the ticks D. silvarum from Buryatia and D. niveus from the Karaganda Region (Central Kazakhstan). The rickettsial genotype RpA4 was most common in the ticks of the genus Dermacentor in Russia and Central Kazakhstan. An analysis of the spread of rickettsias of the STF group shows their close ecological relation to definite types of Ixodes. The rickettsias R. slovaca and RpA4 co-exist in the ticks D. marginatus and D. reticulatus (the western part of a Dermacentor area in Eurasia) and DnS14 and R. sibirica do in D. nuttalli and D. silvarum (the eastern part of the area). D. marginatus and D. reticulatus in the areas characterized by the most specific saturation of a Dermacentor area (the south of West Siberia) are carriers and reservoir of R. sibirica. The rickettsial genotype DnS28 may be now considered to be environmentally associated with one species of ticks--D. nuttalli. At least 6 genotypes of STF rickettsias--R. sibirica, R. astrahan fever (R. conorii), R. slovaca, RpA4, DnS14, DnS28--has been currently identified in Russia and Kazakhstan.

Problems and perspectives of molecular epidemiology of infectious diseases.

Current tendencies and problems of molecular biology and its application in infectious diseases are discussed. Basic stages of the development of molecular epidemiology as a leading tool for epidemiological studies are reviewed. More than 30 years ago, molecular epidemiology was first used to study nosocomial infections. Special attention is given to the achievements of Russian scientists, especially in the discovery of Astrakhan spotted fever Rickettsia, noncultured forms of Vibrio cholerae, and the microorganism "Montezuma." Also, the development of PCR-based methods of identification and typing of Mycobacterium tuberculosis by Russian specialists and its significant role in strategies for diagnosing and treatment of tuberculosis are discussed. Now, the most important field of application of molecular biology methods is bacterial evolution, especially with regard to pathogenic microorganisms and emerging infections. Searching for the novel pathogenic agents, establishment of the infectious nature of diseases with unclear origin, and determining the precise mechanisms of pathogenicity are most intriguing issues. So, molecular biology should play a major role both in clinical and research fields.

[New and emerging rickettsial and bartonella infections]. / Novye i vozvrashchaiushchiesia rikketsiozy i bartonellezy.

There have been recently reports on over 10 new and resurgent rickettsioses and bartonelloses in different countries, which reflects both socioeconomic processes in society and a higher methodological level of indication and identification of causative agents. In 1991, the author' laboratory, N. F. Gamaleya Research Institute of Experimental Medicine, Russian Academy of Medical Sciences, established the etiology of the new rickettsiosis Astrakhan spotted fever. It separated and studied 2 strains of Rickettsia sp. nov. from patients and 8 ones from the carrier the Ixodes tick Rhipicephalus pumilio. It is suggested that the natural focus has transformed to the anthropurgic one due to technogenic environmental pollution. The annual increase in morbidity rates (2000 cases in 1983 to 2000) and its area are a challenge to public health care and medical science. The paper presents data on the new bartonellosis cat-scratch disease (caused by Bartonella henselae) detected not only in Russia. There is also information on tick-borne rickettsiosis, epidemic typhus, and trench fever as resurgent infections.

[Determination of Ehrlichia genome size by pulse gel electrophoresis]. / Opredelenie razmera genoma patogennykh érlikhii metodom pul's-gel'-élektroforeza.

Ehrlichia infections are more and more common in the USA and Europe. The genetics and genome organization of Ehrlichia are little studied due to great difficulties in cultivating these bacteria. Pulse gel electrophoresis was first used to determine the sizes of a genome of 3 representatives of the genus Ehrlichia. The sizes of a genome was established for E. sennetsu (881 kb), for E. risticii (867 kb), E. chaffeensis (1,236 kb).

[Distribution of biological particles in two-phase systems of water soluble polymers and salts]. / Raspedelenie biologicheskikh chastits v dvukhfaznykh sistemakh vodorastvorimykh polimerov i solei.

The paper discusses the problems of purification of Provachek's rickettsias used to prepare typhus vaccines and diagnostic kits. The currently available agents contain many admixtures of rickettsias products, more commonly yolk and chick embryo yolk sack cell detritus, which make it impossible to determine the true antigenic properties of rickettsias in the context of protection and diagnosis. By applying the well-known principle of distribution of biological particles in the two-phase systems of water soluble polymers, investigations were made to examine the distribution of rickettsias and their infected biological mass in the system: polyethylene glycol-6600, sodium-500 dextran sulfate, potassium phosphate, sodium chloride in order to prepare agents of pure rickettsias. It has been found that rickettsias are distributed in the polymer-enriched phase, their number and the degree of purity when isolated from the infected biological mass depends on the capacity of an interphase wherein tissue detritus sorption occurs.

Decreased prevalence of Orientia tsutsugamushi in trombiculid mites and wild rodents in the Primorye region, Far East Russia.

The isolation of Orientia tsutsugamushi was attempted from 249 rodents and approximately 14,000 trombiculid mites captured in the Primorye region, Far East Russia in 1993 and 1994, where high infection rates were recorded in both rodents and mites in the 1960s. However, no rickettsia was isolated from the samples. Low antibody titers against O. tsutsugamushi were detected in 7.1% of the rodents. These results indicate that the prevalence of O. tsutsugamushi in the Primorye region has decreased considerably in the past 30 years.

[The need for a review of the procedure for the preventive vaccination of researchers working with Rickettsia prowazekii]. / Neobkhodimost' peresmotra taktiki profilakticheskoi vaktsinatsii sotrudnikov, rabotaiushchikh s Rickettsia prowazekii.

The analysis of the results of prolonged observations on the prophylactic immunization of employees working with R. prowazekii is presented. The necessity of the differentiated approach to the determination of the immunization schedule and the choice of vaccine is shown. The presence of specific antibodies (Ab) and the level of their titers have been found to be related to the degree of anti-infectious protection. The following characteristics indicate the presence of profound immunological transformation in vaccinees: complement-fixing Ab in titers 1:10 and more and/or immunofluorescent Ab in titers not below 1:180, Ab to protein in the hemagglutination test in titers not below 1:1000. These specific Ab and the level of their titers can be registered after the second injection of live combined typhus vaccine E and the third injection of chemical typhus vaccine. Cases of laboratory infection and their relationship to the character of immunization and the intensity of contacts with R. prowazekii virulent strains are discussed. Attention is drawn to the strict observance of professional safety rules.