Diversity of Bacillus anthracis Strains in Georgia and of Vaccine Strains from the Former Soviet Union

Despite the increased number of anthrax outbreaks in Georgia and the other Caucasian republics of the former Soviet Union, no data are available on the diversity of the Bacillus anthracis strains involved. There is also little data available on strains from the former Soviet Union, including the strains previously used for vaccine preparation. In this study we used eight-locus variable-number tandem repeat analyses to genotype 18 strains isolated from infected animals and humans at different sites across Georgia, where anthrax outbreaks have occurred in the last 10 years, and 5 strains widely used for preparation of human and veterinary vaccines in the former Soviet Union. Three different genotypes affiliated with the A3.a cluster were detected for the Georgian isolates. Two genotypes were previously shown to include Turkish isolates, indicating that there is a regional strain pattern in the South Caucasian-Turkish region. Four of the vaccine strains were polymorphic, exhibiting three different patterns of the cluster A1.a genotype and the cluster A3.b genotype. The genotype of vaccine strain 71/12, which is considered an attenuated strain in spite of the presence of both of the virulence pXO plasmids, appeared to be a novel genotype in the A1.a cluster.     

Prejudice in the former Soviet Union

Research in the United States and Europe has focused on the prejudice of majority groups towards minority groups, the implication somehow being that majority groups were more prejudiced than minority groups. In the former Soviet Union, ethnic environments were more complex; the same ethnic group could be a majority in one region but a minority in others. Using a sample of 1,459 first‐ and fourth‐year university students from eight regions of the former USSR, this study focuses on Russian, Tatar and Ukrainian respondents (n = 821) to test the hypothesis that the status of an ethnic group (majority/minority) or in‐group bias explains members’ prejudice. According to in‐group bias, all ethnic groups are equally prejudiced, minority and majority alike, whereas group status posits that groups in a majority position are more prejudiced. Findings show that group status has greater impact on prejudice than does in‐group bias. This applies, however, only to Russians. Interpretations of the findings rest on Soviet history and the rise of nationalism during the breakup of the Soviet Union.     

Immunobiological properties of 100 years old Bacillus anthracis vaccine strains Lange

The results of the study of B. anthracis vaccine strains Lange 1 and 2 in the form of spores after prolonged storage in 30% glycerine solution at room temperature are presented. The study revealed that spores stored for 100 years germinated and bacterial cells proved to be viable when cultivated in artificial nutrient media and in vivo. They exhibited typical cultural, morphological, biochemical, virulent, antigenic and immunogenic properties.     

Molecular epidemiology of terrestrial rabies in the former Soviet Union

Fifty-five rabies virus isolates originating from different regions of the former Soviet Union (FSU) were compared with isolates originating from Eurasia, Africa, and North America according to complete or partial nucleoprotein (N) gene sequences. The FSU isolates formed five distinct groups. Group A represented viruses originating from the Arctic, which were similar to viruses from Alaska and Canada. Group B consisted of "Arctic-like" viruses, originating from the south of East Siberia and the Far East. Group C consisted of viruses circulating in the steppe and forest-steppe territories from the European part of Russia to Tuva and in Kazakhstan. These three phylogenetic groups were clearly different from the European cluster. Viruses of group D circulate near the western border of Russia. Their phylogenetic position is intermediate between group C and the European cluster. Group E consisted of viruses originating from the northwestern part of Russia and comprised a "northeastern Europe" group described earlier from the Baltic region. According to surveillance data, a specific host can be defined clearly only for group A (arctic fox; Alopex lagopus) and for the Far Eastern part of the group B distribution area (raccoon dog; Nyctereutes procyonoides). For other territories and rabies virus variants, the red fox (Vulpes vulpes) is the main virus reservoir. However, the steppe fox (Vulpes corsac), wolf (Canis lupus), and raccoon dog are also involved in virus circulation, depending on host population density. These molecular data, joined with surveillance information, demonstrate that the current fox rabies epizootic in the territory of the FSU developed independently of central and western Europe. No evidence of positive selection was found in the N genes of the isolates. In the glycoprotein gene, evidence of positive selection was strongly suggested in codons 156, 160, and 183. At these sites, no link between amino acid substitutions and phylogenetic placement or specific host species was detected.     

Identification of Bacillus anthracis strains in China

As part of a project to establish a reference strain collection, phenotypic, biochemical and genetic analyses were carried out on 84 strains of Bacillus anthracis, 81 of them of Chinese origin from various sources. Particular differences from reports on isolates of other origins were the possession of fimbriae and single polar flagella with consequent motility in 77, self-agglutination by 64 and failure to ferment maltose in 60 of the Chinese strains. The findings were considered to be of significance to identification, classification and evolution of B. anthracis.     

Selenium status of pregnant women and newborns in the former Soviet Union

The estimation of the selenium status during pregnancy is of great importance because of the significance of selenium for fetus growth and antioxidant protection of neonates. This problem is especially urgent for Russia and its neighbors because very little data are available and because data on soil selenium predict low intake levels of selenium. A large epidemiological investigation made in various areas of the former USSR allowed us to obtain the first information concerning the subject. Serum samples were obtained during 1990–1998 from 556 female blood donors aged 20–53 yr and 722 pregnant women (18–33 yr) during different times of gestation. The mean serum selenium concentration of nonpregnant women varied from 0.87 μmol/L (Slavutich, Ukraine) to 1.74 μmol/L (Ioshkar-Ola, Mary-El) and that of women at delivery from 0.66 μmol/L (Zaporozie, Ukraine) to 1.34 μmol/L (Sakhalin, Russia). Compilation of literature and present data on serum selenium showed the following relationships: nonpregnant women versus women at delivery, y=x -0.25, r=0.94; women at delivery versus umbilical serum, log y=log x -0.2, r=0.97. The two relationships were used to predict serum selenium values for pregnant women taking into account the progressive serum selenium decrease during advancing pregnancy. In almost half of the towns (i.e. 22–50%), pregnant women were considered to have relative selenium deficiency.     

Genotype diversity of the Bacillus anthracis strains isolated from the Caucasus region

The study of the genotypes of Bacillus anthracis strains isolated from the Caucasus region was performed using MLVA. Among 149 strains, 32 distinctive MLVA-8 genotypes belonging to Ala, A3a, A4 and B1 molecular diversity groups were identified. 9 genotypes were not described previously; 6 genotypes were not found in other geographic regions and could be considered as endemic for Caucasus. The majority of the identified genotypes are widespread not only in Caucasus, but also throughout Eurasia, Africa, and America. Molecular diversity of Caucasian isolates is comparable to the worldwide diversity. This represents historical relations of this region, proximity to ancient trade routes and intensity of the anthrax epizootic in Caucasus. The obtained results are of interest from the theoretical point of view, as well as for the application in epidemiological research of the anthrax outbreaks.     

Virulotypes of Bacillus anthracis strains isolated in Poland

Bacillus anthracis--the causative agent of anthrax--possesses several virulence genes located in the chromosome as well as in two B. anthracis virulence plasmids: pXO1 and pXO2. In the presented study, we determined occurrence of six virulence markers located in the virulence plasmids (capA, capB, capC, pagA, lef and cya) for capsule and toxin production together with virulence-associated gene gerXA and chromosomal gene sap, which are responsible for germination and S-layer biosynthesis respectively. Fourteen strains of B. anthracis isolated in Poland and belonging to five different MLVA genotypes were analyzed by PCR for presence of the aforementioned genes. Two virulotypes were found in tested strains. The only variation was absence of capA, capB and capC due to a lack of pXO2.     

Badger Meles meles diet: a review of literature from the former Soviet Union

We review 69 studies of the diet of Badgers Meles meles L. in the former Soviet Union, using information from 58 Russian-language papers. Animal foods occurred more frequently in the diet than vegetal foods and constituted, on average, 62% of all intake by volume. The most important individual food categories were insects (30% by volume) and small mammals (20% by volume), which appear to be staple foods of Badgers over virtually the whole of the former Soviet Union. Earthworms were mentioned in only a few reports and never exceeded 5% of the diet by volume. Other animal foods included birds, reptiles, amphibians, fish, molluscs and carrion. Fruits were seasonally important in some areas while roots, tubers, bulbs, cereals and nuts were also eaten; but in general, the Soviet literature lacks detailed information about vegetal foods. Many authors comment on seasonal variation in the diet and there was statistically significant seasonal variation in the frequency with which small mammals, reptiles, insects and vegetal foods were eaten. No largescale geographical trends were apparent in the composition of the diet, but foods which were of minor importance overall, such as reptiles, amphibians and molluscs, were sometimes eaten with high frequency in particular localities. The results support the view that Badgers are opportunistic, generalist foragers, with a preference for animal foods when these are available.     

Characterization of Bacillus anthracis strains used for vaccination

Three Bacillus anthracis strains, formerly used as anti-anthrax vaccine strains in Argentina, were characterized from genetic and pathogenic perspectives. Southern blotting and PCR with pXO1 and pXO2 probes and primers, as well as pathogenicity and protection tests in guinea pigs and mice, were performed. Two of the B. anthracis strains contained both pXO1 and pXO2 plasmids, as did the fully virulent strains, while the third was a Sterne-type strain (pXO1+, pXO2-). The three strains were, however, markedly less pathogenic than a wild-type virulent strain. The methodology applied here may be used to characterize other B. anthracis strains.     

Present-day and mid-Holocene biomes reconstructed from pollen and plant macrofossil data from the former Soviet Union and Mongolia

Fossil pollen data supplemented by tree macrofossil records were used to reconstruct the vegetation of the Former Soviet Union and Mongolia at 6000 years. Pollen spectra were assigned to biomes using the plant-functional-type method developed by Prentice et al . (1996). Surface pollen data and a modern vegetation map provided a test of the method. This is the first time such a broad-scale vegetation reconstruction for the greater part of northern Eurasia has been attempted with objective techniques. The new results confirm previous regional palaeoenvironmental studies of the mid-Holocene while providing a comprehensive synopsis and firmer conclusions. West of the Ural Mountains temperate deciduous forest extended both northward and southward from its modern range. The northern limits of cool mixed and cool conifer forests were also further north than present. Taiga was reduced in European Russia, but was extended into Yakutia where now there is cold deciduous forest. The northern limit of taiga was extended (as shown by increased Picea pollen percentages, and by tree macrofossil records north of the present-day forest limit) but tundra was still present in north-eastern Siberia. The boundary between forest and steppe in the continental interior did not shift substantially, and dry conditions similar to present existed in western Mongolia and north of the Aral Sea.     

Immuno-detection of anthrose containing tetrasaccharide in the exosporium of Bacillus anthracis and Bacillus cereus strains

Aims:  Bacillus anthracis strains of various origins were analysed with the view to describe intrinsic and persistent structural components of the Bacillus collagen-like protein of anthracis glycoprotein associated anthrose containing tetrasaccharide in the exosporium.
Methods and Results:  The tetrasaccharide consists of three rhamnose residues and an unique monosaccharide – anthrose. As anthrose was not found in spores of related strains of bacteria, we envisioned the detection of B. anthracis spores based on antibodies against anthrose-containing polysaccharides. Carbohydrate–protein conjugates containing the synthetic tetrasaccharide, an anthrose–rhamnose disaccharide or anthrose alone were employed to immunize mice. All three formulations were immunogenic and elicited IgG responses with different fine specificities. All sera and monoclonal antibodies derived from tetrasaccharide immunized mice cross-reacted not only with spore lysates of a panel of virulent B. anthracis strains, but also with some of the B. cereus strains tested.
Conclusions:  Our results demonstrate that antibodies to synthetic carbohydrates are useful tools for epitope analyses of complex carbohydrate antigens and for the detection of particular target structures in biological specimens.
Significance and Impact of the Study:  Although not strictly specific for B. anthracis spores, antibodies against the tetrasaccharide may have potential as immuno-capturing components for a highly sensitive spore detection system.     

Comparative ecology and epidemiology of lyme disease and tick-borne encephalitis in the former Soviet Union

The recently defined range of human Lyme disease in Russia coincides with that of the intensively studied agent of tick-borne encephalitis (TBE). Although identical vectors transmit both infections during the same seasons and in the same locations, more than twice as many people suffer from Lyme disease as from TBE. Edward Korenberg here describes characteristic features of vectors with respect to their seasonal activity, abundance, rates of infection by TBE and Lyme disease agents, as well as incidence of human-tick contact, and how these factors determine the incidence of TBE and Lyme disease in different regions.     

Filing of microorganism strains for patent claims in the Soviet Union

The modern state of depositing microorganisms with regard to patent procedure is considered. The main stages of depositing are described. Two questions of paramount importance for depositors are examined, i.e. what kind of microorganisms should not be deposited with regard to patent procedure, who and on what terms can get samples of deposited microorganisms from the collection. A number of recommendations for researchers potential depositors of microorganisms, are given.     

Strategy for identification of Bacillus cereus and Bacillus thuringiensis strains closely related to Bacillus anthracis

Bacillus cereus strains that are genetically closely related to B. anthracis can display anthrax-like virulence traits (A. R. Hoffmaster et al., Proc. Natl. Acad. Sci. USA 101:8449-8454, 2004). Hence, approaches that rapidly identify these "near neighbors" are of great interest for the study of B. anthracis virulence mechanisms, as well as to prevent the use of such strains for B. anthracis-based bioweapon development. Here, a strategy is proposed for the identification of near neighbors of B. anthracis based on single nucleotide polymorphisms (SNP) in the 16S-23S rRNA intergenic spacer (ITS) containing tRNA genes, characteristic of B. anthracis. By using restriction site insertion-PCR (RSI-PCR) the presence of two SNP typical of B. anthracis was screened in 126 B. cereus group strains of different origin. Two B. cereus strains and one B. thuringiensis strain showed RSI-PCR profiles identical to that of B. anthracis. The sequencing of the entire ITS containing tRNA genes revealed two of the strains to be identical to B. anthracis. The strict relationship with B. anthracis was confirmed by multilocus sequence typing (MLST) of four other independent loci: cerA, plcR, AC-390, and SG-749. The relationship to B. anthracis of the three strains described by MLST was comparable and even higher to that of four B. cereus strains associated with periodontitis in humans and previously reported as the closest known strains to B. anthracis. SNP in ITS containing tRNA genes combined with RSI-PCR provide a very efficient tool for the identification of strains closely related to B. anthracis.     

Insight into the studies of the practical use of microalgae in the former Soviet Union

Microalgae have been intensively cultivated and used in the national economy of the Soviet Union in Russia (Moscow, Leningrad, Pushcino, Krasnoyarsk, Irkutsk), the Ukraine, Byelorussia (Minsk), Azerbaijan (Baku), Uzbekistan (Tashkent), Latvia (Riga) and other former Soviet republics. The first studies on the practical use of microalgae started already at the beginning of last century and were connected with treatment of waste water in biological ponds. Ecophysiological studies of marine plankton algae in mono- and mixed cultures as well as in experimental systems with zooplankton organisms started at the end of the 1950s. In the 1960s, started to apply different equipments for cultivation of microalgae in open water basins and closed ecological systems. Research innovations were introduced in agriculture. The productivity of aquatic ecosystems and the distribution of energy within food chains were studied. Intensive research on microalgae continued, paying attention to the chemical composition, physiological and biochemical features. Nutrient supply appeared as one of the main problem for productivity of algal cultures, along with the procession, biochemistry of nutrients and nutrition value of organic matter produced by algae. The most attention was paid to the Chroococcales proteins and possibilities for their use in man-made closed ecological systems. The potential use of microalgae for regeneration of the atmosphere and purification of water in spacecrafts was studied. During the 1970s attention was paid to the role of algae in self-purification processes and biodegradation of surface-active substances in water bodies. Research in microalgae as bioindicators for the assessment of environment quality of aquatic ecosystems and as test objects for biotesting of natural and waste waters was conducted from the 1970s to 1980s.     

Genetic and taxonomic diversity of the house mouse Mus musculus from the asian part of the former Soviet Union

Genetic diversity of the house mouse Mus musculus from 12 local populations (n = 65) of the central and eastern parts of the former Soviet Union was examined using RAPD-PCR. About 400 loci were identified, encompassing approximately 500 kb of the mouse genome. Genetic diversity was assessed using NTSYS, POPGENE, TFPGA, and TREECON software programs. In general, the house mouse sample from the regions examined was characterized by moderate genetic variation: polymorphism P = 95.6%, P99 = 60.7%, P95 = 24.2%; heterozygosity H = 0.089; the mean observed number of alleles n(a) = 1.97; effective number of alleles n = 1.13; intrapopulation differentiation deltaS = 0.387; gene diversity h = 0.09. Individual local populations displayed different levels of genetic isolation: the genetic subdivision index G(st) varied from 0.086 to 0.324 at gene flow Nm varying from 5.3 to 1.05, while the interpopulation genetic distance D(N) ranged from 0.059 to 0.186. Most of the genetic diversity of the total sample resided within the local populations: H(S) = 0.6, total gene diversity H(T) = 0.09. The exact test for differentiation, however, did not confirm the affiliation of all the mice examined to one population: chi2 = 1446, d.f. = 724, P = 0.000. Molecular markers specific to four subspecies (musculus, castaneus, gansuensis, and wagneri) were identified. Moreover, in some cases the populations and individual animals exhibited traits of different subspecies, suggesting their introgressive hybridization. It was demonstrated that the house mouse fauna on the territories investigated was characterized by the prevalence of musculus-specific markers, while gansuensis-specific markers ranked second. The castaneus-specific markers were highly frequent in the Far East, but almost absent in Central Asia, where wagneri-specific markers were detected. It was suggested that house mice from Turkmenistan could belong to one of the southern subspecies, which had not deeply penetrated into the Asian fauna of the former Soviet Union. In phenogenetic (UPGMA) and phylogenetic (NJ) reconstructions this form with the high bootstrap support was placed at the tree base, while the isolation of other clusters was not statistically significant. It is thus likely that the house mice from Turkmenistan are closest to the ancestral form of the genus Mus on the territory of the former Soviet Union.