[O3:K6 serogroup vibrio parahaemolyticus - the causative agent of food toxic infection outbreaks in Primorsky region of Russian federation].

AIM: Comparative evaluation of biological properties of parahemolytic vibrios that had determined outbreaks and sporadic cases of food toxic infection in Primorsky Region in 2012 and previous years. Materials AND METHODS: 40 clinical strains of Vibrio parahaemolyticus isolated in 2012 were studied in comparison with 62 strains from this region that had been characterized by us previously. Virulence was evaluated by a complex method: hemolytic activitywas determined in Kanagawa test (KT), urease - in Kristensen medium. Serotyping was carried out by a commercial kit of O/K sera. PCR-genotyping was carried out by marker genes of 7 pathogenicity "islands" (VPaI-1-7). RESULTS: All the strains isolated from patients in 2012 had KT-positive and urease-negative phenotype, belonged to O3:K6 serogroup and contained marker genes of 7 VPal that allowed to consider them members of a "pandemic" clone as the other clinical strains from this region. However among 2012 strains an increase of number of antibiotic-resistant variants was established compared with 1997 isolates. CONCLUSION: The data obtained give evidence on the risk of spread of a "pandemic" clone of V. parahaemolyticus in the Far-Eastern region of Russia, a dangerous tendency of antibiotic-resistant variant formation and a necessity to monitor morbidity and the environment with mandatory PCR-detection of genes associated with virulence including integrated into pathogenicity "islands".

[Collection of Vibrio parahaemolyticus species members: phenotypic and genotypic characteristics].

AIM: Formation of Vibrio parahaemolyticus collection according to modern methodical opportunities and understanding of causative agent biology. MATERIALS AND METHODS: Traditional biochemical tests and PCR-testing of species-specific genes were used to confirm species membership. Catalase, DNAse, proteolytic and tweenase activity was determined by common methods. Virulence was evaluated by a complex method: hemolytic activity was determined in Kanagawa test (KT), urease--in Christensen medium, PRC-testing of tdh and-trh genes. Serotyping was carried out with a commercial O/K-sera kit. PCR-genotyping was carried out by marker genes of 7 pathogenicity islands (VPaI-1-7). RESULTS: Species membership was confirmed for the studied strains. Serologic typing allowed to detect members of 18 serologic groups among the collection strains. All the collection cultures were divided into 4 groups based on KT-Ure-tdh-trh features recombination. A number of genetic variants were detected, strains belonging to a pandemic group and O3:K6 serogroup were determined. CONCLUSION: A collection of V. parahaemolyticus cultures was formed and characterized by a large set of pheno- and genotypic features. A database was developed including information on strain origins, pheno- and genetic features, with genetic variants given, for ease of use of the collection.

[The differentiation of specimiens of Vibrio parahaemolyticua and Vobrio alginolyticus].

Vibrio parahaemolyticua and Vobrio alginolyticus are phylogenetically closely-related species. They have common ecological niches, same cultural features and similar biochemical characteristics. The phenotype variability and taxonomy similarity of strains of these species impedes differentiation of Vibrio parahaemolyticua and Vobrio alginolyticus according biochemical characteristics. To obtain reliable results of diagnostic application of additional methods of differentiation and identification these two species of bacteria are needed. The study was organized to comparatively evaluate effectiveness of biochemical testing, polymerase chain reaction analysis and mass-spectrometry technique in differentiation of species of Vibrio parahaemolyticua and Vibrio alginolyticus. The study implemented analysis of methods of differentiation of species of Vibrio parahaemolyticua and Vibrio alginolyticus using model of collection including atypical strains of these species. To substantiate species belonging of strains of Vibrio parahaemolyticua and Vobrio alginolyticus such techniques are to be additionally applied to biochemical methods of identification as polymerase chain reaction analysis with species-specific primers of genes of metalloproteinase (collagenase) vppC and vapC. The MALDI-TOFF method of mass-spectrometry can be used as additional effective method of identification and inter-species differentiation of species of Vibrio parahaemolyticua and Vibrio alginolyticus isolated from various sources.

[The development of complex technique of evaluation of virulence of parahemolytic vibrio].

The article deals with results of studying parahemolytic vibrio separatedfrom different sources according their phenotype and genotype attributes associated with virulence. In certain cases the mismatch of results of Kanagava tests and polymerase chain reaction test of gene tdh was established. The need in virulence complex evaluation is substantiated. This complex has to include detection of hemolytic activity in Kanagava test and urease activity on the Kristensen medium and polymerase chain reaction detection of genes tdh and trh. The developed complex technique is described. The formula of pathogenic strains is established Three alternatives of virulent parahemolytic vibrio are given. The test-strains Vibrio parahaemolyticus are proposed as control in testing phenotype and genotype strains according virulence signs.

[Study of possibility of formation of Vibrio parahaemolyticus pandemic clones based on retrospective PCR-screening of clinical strains].

AIM: PCR-genotyping of Vibrio parahaemolyticus strains that had caused sporadic diseases in Novorossiysk from 1973 to 1976. MATERIALS AND METHODS: 24 clinical strains of V. parahaemolyticus isolated in Novorossiysk, most of which belonged to serogroups O4:K12 and O4:K8; 10 O3:K6 strains--causative agents of gastroenteritis outbreak in Vladivostok (1997) and 3 from Japan (1971) were used. PCR genotyping was performed by a set of marker genes of 7 pathogenicity islands (VPaI-1 - VPaI-7) and a number of other pathogenicity factors. RESULTS: All the strains isolated in 1970s differed significantly by sets of VPaI marker genes. In contrast to causative agents of outbreak in Vladivostok that contain all 7 VPaI genes (that is, members of the pandemic group that had spread globally since 1996) none of the O4:K12 and O4:K8 Novorossiysk strains contained the full set of all the VPaI genes. However this set was distributed among the members of the group. CONCLUSION: Taking into account that O4:K12 and O4:K8 serogroups are considered by a number of authors as O3:K6 serovariants, PCR-screening data obtained by us allows to assume that horizontal transfer of mobile elements (VPaI) between strains circulating in the region could have led to the formation of pandemic clones already in the 1970s. This implies that in several coastal regions in certain periods of time conditions that favor these process may form, and risk of infection with pandemic clones is associated not only with import of seafood.

[Characteristics of Vibrio cholerae nonO1/nonO139 serogroup strains that caused diseases in population of Rostov region].

AIM: Genotype characteristic and determination of serological properties of Vibrio cholerae nonO1/nonO139 strains that caused diseases in population of Rostov region from 2000 to 2009. MATERIALS AND METHODS: 15 clinical strains of V. cholerae nonO1/nonO139 were studied. Serotyping was performed by using a kit of monospecific typing sera against serogroup 02-084 cholera vibrios obtained from Rostov Research Institute for Plague Control, PCR and VNTR-genotyping--by using specific primers described in scientific publications and constructed by us. RESULTS: Serologic features of strains are very diverse and strains contain various combination of pathogenicity factor genes that seem to be interchangeable. Similar pattern was observed for VNTR-genotyping. Distribution of the examined strains by VNTR-genotyping did not correlate with either PCR-genotyping data or serotyping, or place and time of isolation. CONCLUSION: The data obtained indicates a lack of general source of human infection even in the same location and time period. On the other hand, serological and genotypic features of V. cholerae nonO1/nonO139 may undergo changes in the process of staying in the macro organism or environment due to high plasticity of their genome.

[New nutrient medium for the cultivation and isolation of the plague microbe ChDS-37 as an element of the mobilization reserve of specialized antiepidemic teams of the Russian Inspectorate for the Protection of Consumer Rights and Human Welfare].

A new nutrient medium has been designed to culture and isolate the plague microbe ChDS-37 on the basis of the pancreatic digest of baker's yeast. The results of laboratory tests of the designed medium, by using 10 plague microbe strains and those of approval during the tactical and special training of a specialized antiepidemic team (SAET), suggest that the medium has some advantage over reference media and creates prerequisites for being incorporated into the mobilization reserve of a SAET.

[Relation between pulmonary ventilation parameters, exercise tolerance and quality of life in patients with chronic obstructive lung disease]. / Vztah ventilacních plicních parametru, tolerance fyzické záteze a kvality zivota u pacientu s chronickou obstrukcní plicní nemocí.

Patients with chronic obstructive pulmonary disease (COPD) do usually have decreased tolerance of exercise capacity and impaired quality of life. Several studies have shown that exercise capacity is related relatively weakly to lung functions in this group of patients. The aim of the present study was to find parameter which could better reflect or predict maximal exercise capacity. 19 patients with the diagnosis COPD with mean value of forced expiratory volume in one second (FEV1) 46% predicted (range 21-79%) entering pulmonary rehabilitation program were included into the study. Enrolled patients were chosen to cover the whole range of airway obstruction severity. Post-bronchodilator static and dynamic ventilation parameters were used for evaluation and calculation. Quality of live was measured using St. George's respiratory questionnaire (SGRQ), evaluating symptoms, activity and impact of the disease with range from 0 (the best level) to 100 (the worst level). Values of FEV1 (p < 0.001) and ratio of FEV1 to vital capacity (FEV1/VC, p < 0.001) were significantly positively correlated with 6 minute walking distance (6MWD). FEV1/VC were closely related to 6MWD then FEV1. The degree of hyperinflation expressed by residual volume (RV, p < 0.005) and by ratio of residual volume to total lung capacity (RV/TLC, p < 0.001) significantly negatively correlated with 6MWD. Maximal occlusion mouth pressures (PImax, p < 0.05) were positively related to 6MWD. Total score of SGRQ correlated significantly to maximal exercise capacity. Pulmonary function tests and respiratory muscle function have important impact on exercise tolerance in patients with COPD. Tolerance of exercise capacity is significantly reflected by total score of quality of life in this group of patients.

[Genotyping of Yersinia pestis: variability of locus (CAAA)n in natural strains isolated in territory of former SSSR]. / Genotipirovanie Yersinia pestis: variabel'nost' lokusa (CAAA)n u prirodnykh shtammov, vydelennykh na territorii byvshego SSSR.

Genome polymorphism by the locus (CAAA)n was studied in 69 strains of Yersinia pestis isolated from natural foci of the former Soviet Union. The polymorphism was found to be represented by ten alleles in chromosomes, which could be regarded as evidence of variability of this VNTR-locus (diversity index, DI = 0.86). The value of DI was found to vary substantially: from 0.24 in a group of vole strains from seven isolates from the Transcaucasian highlands to 0.77 in nine strains from the Central Asia desert focus. The allele polymorphism of the variable locus (CAAA)n in natural strains of Y. pestis was suggested to be used as a possible genetic marker of the strain. It was concluded that the oligonucleotide primers used in polymerase chain reaction should be upgraded to the genotyping accuracy.

[Analysis of toxin genetic determinants of Vibrio cholerae virulence cassette and neuraminidase with DNA probes]. / Izuchenie geneticheskikh determinant toksinov kassety virulentnosti i neiraminidazy kholernykh vibrionov s pomoshch'iu molekuliarnogo zondirovaniia.

V. cholerae strains of different origin have been studied for the presence of cholera toxin genes (vct), the proximal part of the virulence cassette including genes zot, ace and orfU, as well as neuraminidase genes (neu), in their genomes with the use of molecular DNA probes. The possibility, in principle, for some strains to lose only a part of their virulence cassette (gene vct), while retaining its proximal part has been shown. In most cases such strains are isolated from patients with diarrhea of different severity and may probably play some etiological role, provided that the expression of the genes of additional toxins of the virulence cassette occurs. The gene expressing neuraminidase which facilitates the penetration of cholera toxin into the epithelial cells of the intestine is always present in vct+ strains and may be absent in vct- strains. The absence of genetic relationship between neuraminidases in V. cholerae O139 and V. cholerae O1 and non-O1 (non-O139) has been confirmed. The problems in connection with the integration and deletion of genetic determinants of V. cholerae virulence factors are discussed.

Vascular and nerve disorders at exposure to hand-held vibrating tools related to ISO 5349.

The authors present the results of the investigation of peripheral nerves of the hand in a group of 30 glass cutters, 21 metal grinders, 10 blacksmiths and 24 workers who worked with nailers. The measurements of vibrotactile sensitivity were carried out by Vibrometry System Brüel & Kjaer Type 9627. The vibrotactile perception thresholds (VPT) were measured in frequency range from 8 to 500 Hz. The same investigations were carried out in 103 healthy subjects with no previous work-exposure to vibration or shocks. Further electromyographic examination was carried out in all members of exposed groups. The results of measurements VPT have proved reliability and high sensitivity of this method for the early detection of light disorders of peripheral nerves of the hand due to vibration. All the workers also passed investigation of vascular system of fingers. Simultaneously the measurement of vibration and shocks of single types of hand-held tools used by workers was done. The results of measurement have been worked up according to ISO 5349 (1) that enables to determine the beginning of the incidence of disorders of the hands and arms on the base of measured values of vibration and shocks. The results of the vascular system investigation have been compared with the results of the assessment of exposure according to ISO 5349. The conformity has been proved in the case of hand-transmitted vibrations not in the case of shocks.

[Studies on halophilic vibrios causing a food poisoning outbreak in the city of Vladivostok]. / Galofil'nye vibriony, obuslovivshie vspyshku pishchevoi toksikoinfektsii vo Vladivostoke.

V. parahaemolyticus and V. alginolyticus strains isolated from patients during an outbreak of an acute enteric disease in Vladivostok in 1997 were studied. All strains were found to possess typical taxonomic signs. V. parahaemolyticus isolated from humans had direct heat stable haemolysin exotoxin. The overwhelming majority of these strains belonged to serovar O3K6. Among the cultures under study 7 phage types were determined: phage types 1, 2, 7, 10 in 8 V. parahaemolyticus strains and phage types 2, 4. 5. 7 in 5 V. alginolyticus strains. The diagnostic halophilic phage lyzed vibrios in 30.2% of strains. The cultures under study were found to be highly sensitive to chloramphenicol, cefotaxime, nalidixic acid and cyprofloxacin. The study proved that the outbreak of alimentary toxicoinfection was caused by vibrios of serogroup O3:K6.

[Cystic fibrosis--a disease of adolescents and adults?]. / Cystická fibróza--nemoc dospívajících a dospelých?

BACKGROUND: Cystic fibrosis (CF) is no longer a childhood disease. Since the identification of the gene in 1989 research has made advances and changed views on the pathogenesis, diagnosis and treatment. The objective of the present work is to make doctors treating adult patients familiar with modern therapeutic methods and their value. METHODS AND RESULTS: In the CF Centre of the Faculty Hospital in Prague Motol 349 patients are followed up on a long-term basis, incl. 95 who died since 1985. Hundred and twenty six (36.1%) patients survived to the age of 18 years, of those 41 died and 85 patients live. Comparison of semilongitudinal data of a group of 83 patients born before 1975 whose treatment during childhood and puberty was inadequate and 196 patients born in 1976-90 treated by modern methods proved the great effect of treatment on the course and prognosis of the disease. The median age at death increased during from 12.2 years in 1985-90 to 18.8 years in 1991-1998 (p = 0.004). The nutritional status of adult patients is satisfactory in 40.4%, poor in 33.3% and marginal in 26.3%. A normal pulmonary function was recorded in 17.5%, 22.8% are severely affected, the majority of patients (59.7%) has values within 40 to 80% of normal levels. CONCLUSIONS: Modern intensive treatment improved the prognosis and quality of live in patients with CF. Critical deterioration of the clinical condition shifted to the threshold of adult age. It is therefore essential that doctors treating such patients should be familiar with this issue.

[Evaluation of antibiotic sensitivity of pathogenic vibrios of various species]. / Otsenka chuvstvitel'nosti k antibiotikam patogennykh vibrionov raznykh vidov.

The recent increase of the number of antimicrobials and isolation of antibiotic resistant strains from humans and environmental objects is indicative of the necessity of further investigation of antibiotic susceptibility of the representatives of the genus Vibrio pathogenic for man to provide rational therapy of the diseases due to them. Susceptibility of 160 strains of pathogenic vibrios of 9 species to 11 antibiotics and chemotherapeutic drugs was assayed by the method of serial dilutions in agar media. The isolates were shown to be highly susceptible to chloramphenicol, doxycycline, cefotaxime, nalidixic acid and ciprofloxacin which made it possible to consider them as the drugs of choice in the treatment of the diseases caused by the microorganisms. A tendency to form polyantibiotic resistant strains within every species of tested pathogenic vibrios was observed. It conditioned the prospects of further profound study of the phenomenon with the analysis of the genetic determination of antibiotic resistance markers in pathogenic vibrios.

[New serological variants of Yersinia enterocolitica and Yersinia kristensenii]. / Novye serologicheskie varianty Yersinia enterocolitica i Yersinia kristensenii.

Seven Yersinia strains, selected from a number of cultures, not agglutinated by 32 typing sera, were found to have original antigenic features. These strains were given over to the Tarasevich State Research Institute for Standardization and Control of Medical Biological Preparations and registered under O-factor formulae and numbers: Y. kristensenii 165-036, Y. enterocolitica 170-037, Y. enterocolitica 168-038, Y.enterocolitica 167-039, Y. enterocolitica 166-040, Y. enterocolitica 169-041, Y. kristensenii 171-042. In our collection of strains antigenic analogs of new typing strains were found. The inclusion of sera to new serovars into the set of Yersinia diagnostic sera will widen the possibilities of epidemiological analysis.

[Choice of commercial elective nutrient media for the isolation of pathogenic vibrios of different species]. / Podbor kommercheskikh élektivnykh ligatel'nykh sred dlia vydeleniia patogennykh vibrionov raznykh vidov.

Commercial nutrient media and their modifications are assessed, including elective differentiation medium for V. cholerae developed at the Rostov Research Anti-Plague Institute for the isolation of pathogenic vibrios. V. cholerae cholerae P-1 (145), V. cholerae el tor M-878, V. cholerae non 01 P-9741, E. coli 18, P. vulgaris 19, and 48 strains of vibrios belonging to different species were used in the study. All the strains used were characterized as to their nutritive requirements. Alkaline agar manufactured by the I. I. Mechnikov Research Institute of Vaccines and Sera was found to be the optimal medium for the isolation of pathogenic vibrios, for it provided the growth of all the examined strains from the inoculation dose of 100 bacterial cells. Addition of potassium tellurite to this medium in a dose of 30 to 50 mg/ml improved its selective properties. Elective differential medium for V. cholerae, though somewhat inferior to alkaline agar in sensitivity, provided the growth of colonies of the overwhelming majority of the strains and inhibited the growth of associated bacteria. Hence, these two media are recommended as elective media for the diagnosis and selection of pathogenic vibrio. Endo and Ploskirev's media with alkaline agar in 1 to 1 ratio may be recommended as additional ones for the isolation of vibrios from patients and from environmental objects.

[Vibrios pathogenic to humans and laboratory diagnosis of diseases caused by them]. / Patogennye dlia cheloveka vibriony i laboratornaia diagnostika vyzyvaemykh imi zabolevanii.

Present-day taxonomic status of vibrios pathogenic for humans is discussed, as are the phenotypical characteristics of some species, factors of virulence, and clinical manifestations of diseases caused by these agents. A scheme of isolation and identification of vibrios pathogenic for humans is offered.

[The use of a cholera coagulating diagnostic agent under the cholera epidemic conditions in Dagestan]. / Primenenie kholernogo koagliutiniruiushchego diagnostikuma v usloviiakh épidemii kholery v Dagestane.

Dried cholera diagnosticum for the slide coagglutination test was obtained. The diagnosticum, found to be highly active and specific, permitted the detection of Vibrio cholerae in the analyzed material at a concentration of 10(6)-10(8) microbial cells/ml. The diagnosticum was used during cholera epidemic in Daghestan for the detection and rapid identification of cholera vibrios. In all cases the positive results of the coagglutination test were confirmed by other investigation methods (no cases of hyperdiagnosis was registered). The production of these diagnostica in small batches on the basis of specialized research laboratories was recommended.

[The properties of the Vibrio cholerae strains isolated in large districts in western Dagestan in 1994]. / Svoistva shtammov Vibrio cholerae, vydelennykh v krupnykh raionakh na zapade Dagestana v 1994 g.

190 V. cholerae cultures isolated by the specialized antiepidemic brigade of the Rostov-on-Don Research Institute for Plague Control in the Khasavyurt, Babayurt and Novolaksk regions of Daghestan in August-October 1994. All isolated strains were typical with respect to their morphological and cultural properties and could be agglutinated (with the exception of one strain) to the titer or half-titer with diagnostic cholera serum and Ogawa serum. 4 strains had signs of RO-dissociation, 4 strains were agglutinated with Inaba serum in a low titer. All strains were resistant to diagnostic bacteriophages. Cyproxin and doxicycline proved to be the most active agents for the treatment of patients. Agglutinins, vibriocidins and antidermonectrotic antibodies in diagnostic titers were detected in the sera of all patients and Vibrio carriers.

Impulsive vibration and exposure limit.

Several cases of exposure to hand-transmitted shocks in two different factories were studied with an intention to improve our knowledge about the influence of the impulsive components of vibration on human being. Two types of shocks affecting the workers could be distinguished, i.e., shocks having small amount of energy at frequencies higher than 100 Hz and shocks having relatively high frequency components up to 1000 Hz. An exposure to the hand-arm shocks of both types was measured and assessed in accordance with the international standard ISO 5349. Health condition of workers was examined. Carpal tunnel syndrome was found among the workers of the workshop in wood industry, who work with staple gun. The investigation of the health condition of about 60 forge workers up to now did not confirm the severity of hand-arm shocks as it had been assessed according to the Annex A of the ISO 5349. The exposure to hand-arm shocks in the forge leads to the incidence of occupational diseases and disorders, which however, differ from the diseases normally reported as a consequence of the hand-arm vibration and shocks. It seems obvious that for the assessment of shock exposure, the ISO 5349 is not quite suitable in its present form.