RESUMO
72 clinical strains of Klebsiella spp. isolated from samples obtained from humans in Novosibirsk, Russia, were analyzed. Species identification of strains was performed using 16S rRNA and rpoB gene sequences. It was revealed that Klebsiella pneumoniae strains were dominant in the population (57 strains), while the remaining 15 strains were K. grimontii, K. aerogenes, K. oxytoca and K. quasipneumoniae. By molecular serotyping using the wzi gene sequence, K. pneumoniae strains were assigned to twenty-one K-serotypes with a high proportion of virulent K1- and K2-serotypes. It was found that K. pneumoniae strains isolated from the hospitalized patients had a higher resistance to antibiotics compared to the other Klebsiella species. Real-time PCR revealed that the population contained genes of the blaSHV, blaTEM, blaCTX families and the blaOXA-48 gene, which are the genetic determinants of beta-lactam resistance. It has been shown that the presence of the blaCTX sequence correlated with the production of extended-spectrum beta-lactamases, and phenotypic resistance to carbapenems is due to the presence of the blaOXA-48 gene. At the same time, the carbapenemase genes vim, ndm, kpc, imp were not detected. Among the aminoglycoside resistance genes studied, the aph(6)-Id and aadA genes were found, but their presence did not always coincide with phenotypic resistance. Resistance to fluoroquinolones in the vast majority of strains was accompanied by the presence of the aac(6')-IB-cr, oqxA, oqxB, qnrB, and qnrS genes in various combinations, while the presence of the oqxA and/or oqxB genes alone did not correlate with resistance to fluoroquinolones. Thus, the detection of blaCTX and blaOXA-48 can be used to quickly predict the production of extended-spectrum beta-lactamases and to determine the resistance of Klebsiella to carbapenems. The detection of the aac(6')-Ib-cr and/or qnrB/qnrS genes can be used to quickly determine resistance to fluoroquinolones.
RESUMO
The evaluation of content of DNA of lactobaccilli and particular types of aerobic anaerobic opportunistic bacteria in sampling of scrapes from urogenital tract offemale patients of the network laboratory INVITRO was implemented. The technique of polymerase chain reaction in real-time was implemented. It is demonstrated that decreasing of content of lactobaccilli in total bacterial mass isfollowed by increasing of occurrence, concentration and relative content of all types of opportunistic pathogens except ureaplasmna. These changes are expressed in different degree for different types of opportunistic pathogens. The increasing of varieties of types of microflora of urogenital tract under decreasing of content of lactobaccilli is noted.
Assuntos
Bactérias Aeróbias/isolamento & purificação , Bactérias Anaeróbias/isolamento & purificação , Sistema Urogenital/microbiologia , Bactérias Aeróbias/classificação , Bactérias Aeróbias/patogenicidade , Bactérias Anaeróbias/classificação , Bactérias Anaeróbias/patogenicidade , Feminino , Humanos , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Sistema Urogenital/patologiaRESUMO
Ticks of the genus Ixodes were collected in 2010 in the lowland part of Toguchinsk district of Novosibirsk Province (Russia) and in the forest-park area of Novosibirsk Scientific Centre and its outskirts (Sovetskiy district of Novosibirsk), and identified as Ixodes persulcatus (Schulze, 1930) (18 females and 13 males) and Ixodes pavlovskyi (13 females and 10 males). Ten specimens of each sex from each collecting site were examined. The following nine characters were used: the length and width of the scutum (conscutum) and of the gnathosoma in ventral view; the length of palpal segments II-III; the width of the hypostome; the length of idiosoma with scapula, of leg I, of the medial spur on fore coxa (Taiga..., 1985; Filippova, Musatov, 1996; Filippova, Panova, 1998). According to morphometric characters, specimens of Ixodes pavlovskyi collected in the forest-park area of the Novosibirsk Scientific Centre were identified as the subspecies I. p. occidentalis Filippova et Panova, 1998. Nucleotide sequences of the COI mitochondrial gene fragment were determined for 56 ticks. Phylogenetic analysis of the COI gene fragment in representatives of the persulcatus-ricinus species-group dwelling in Asia demonstrated high degree of conservatism. Molecular-genetic methods allow reliable identification of morphologically similar species I. pavlovskyi and I. persulcatus, pathogenic for humans.
Assuntos
Proteínas de Artrópodes/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Ixodes/classificação , Ixodes/genética , Filogenia , Animais , Feminino , Humanos , Ixodes/anatomia & histologia , Ixodes/enzimologia , Masculino , Sibéria , Especificidade da EspécieRESUMO
The definition and molecular typing of Borrelia miyamotoi transmitted by the Ixodes persuccatus ticks was based on the partial sequencing of the 16S rRNA, p66, and glpQ genes. All the B. miyamotoi analyzed sequences of the 16S rRNA, glpQ, and p66 gene fragments from I. persulcatus were identical and had 99.9-100% similarity to corresponding genes of the B. miyamotoi strain FR64b isolated in Japan. The analyzed amino acid sequences revealed that the 66 protein B. miyamotoi in the site corresponding to the surface-exposed domain contained considerable difference from the Borrelia hermsii, the typical member tick-born relapsing fever, as from Borrelia lonestari transmitted by the Ixodes ticks.
Assuntos
Proteínas de Bactérias/genética , Borrelia/genética , Ixodes/microbiologia , Doença de Lyme/microbiologia , Diester Fosfórico Hidrolases/genética , Porinas/genética , RNA Ribossômico 16S/genética , Sequência de Aminoácidos , Animais , Borrelia/classificação , Borrelia/isolamento & purificação , Humanos , Japão , Dados de Sequência Molecular , Filogenia , Febre Recorrente/microbiologia , Federação RussaRESUMO
Unfed adult Ixodes persulcatus ticks from five regions of Russia were examined to analyze the distribution and diversity of Borrelia miyamotoi. DNA of B. miyamotoi was found in 1.8% of ticks from Leningrad Oblast, 2.9% from Sverdlovsk, 4.5% from Novosibirsk, 2.3% from Irkutsk Oblast, and 2.5% from Khabarovsk Krai. The molecular typing of the B. miyamotoi DNA was based on the partial sequencing of the 16S rRNA, p66, and glpQ genes. The only genetic variant of B. miyamotoi was detected in all samples of ticks collected from these five territories.
Assuntos
Vetores Aracnídeos/microbiologia , Infecções por Borrelia/microbiologia , Borrelia/classificação , Borrelia/isolamento & purificação , Ixodes/microbiologia , Animais , Proteínas de Bactérias/genética , Borrelia/genética , Infecções por Borrelia/epidemiologia , Contagem de Colônia Microbiana , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Monitoramento Ambiental , Monitoramento Epidemiológico , Humanos , Dados de Sequência Molecular , Diester Fosfórico Hidrolases/genética , Filogenia , Densidade Demográfica , Porinas/genética , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Federação Russa/epidemiologia , Análise de Sequência de DNARESUMO
There has been recently a rise in referrals for Ixodes tick bites in the spring and summer periods in the Kamchatka Territory. Among the dominant tick species, there has been the taiga tick Ixodes persulcatus habiting the extensive areas of the southern and central parts of the peninsula. Examination of 84 I. persulcatus females collected from human beings and domestic animals in 2003 to 2007 detected DNA of the pathogens of tick-borne borreliosis (B. burgdorferi sensu lato), rickettsiasis (R. tarasevichiae and R. helvetica), and Ehrlichiosis/anaplasmosis (A. phagocytophilum). Tick-borne encephalitis RNA and antigens and babesiasis DNA were not found in the study samples. Despite the small number of taiga ticks in Kamchatka, the detection of the pathogens of various infectious diseases in the ticks suggests that there may be a risk for contamination of the peninsula's population with these pathogens.
Assuntos
Borrelia/isolamento & purificação , Vírus da Encefalite Transmitidos por Carrapatos/isolamento & purificação , Ixodes/microbiologia , Reação em Cadeia da Polimerase , Anaplasma/genética , Anaplasma/isolamento & purificação , Animais , Babesia/genética , Babesia/isolamento & purificação , Babesiose/epidemiologia , Proteínas da Membrana Bacteriana Externa/análise , Proteínas da Membrana Bacteriana Externa/genética , Borrelia/genética , Infecções por Borrelia/epidemiologia , DNA Bacteriano/análise , DNA de Protozoário/análise , Ehrlichiose/epidemiologia , Vírus da Encefalite Transmitidos por Carrapatos/genética , Encefalite Transmitida por Carrapatos/epidemiologia , Monitoramento Ambiental , Monitoramento Epidemiológico , Feminino , Humanos , Ixodes/virologia , RNA Viral/análise , Rickettsia/genética , Rickettsia/isolamento & purificação , Infecções por Rickettsia/epidemiologia , Sibéria/epidemiologiaRESUMO
In our study, Borrelia were revealed in the taiga ticks Ixodes persulcatus collected on vegetation by flagging, as well as in the ticks removed from the people who asked for help in the vaccination center located in the Novosibirsk Scientific Center of the Siberian Branch of Russian Academy of Science (NS SB RAS). By the isolation of Borrelia on BSK-H medum, the occurrence of B. garinii, B. afzelii, and B. miyamotoi was established in the territory of NSC. B. miyamotoi isolates were unstable and lost their ability to growth in later passages. DNA of the same three species of Borrelia was detected by PCR in the samples of ticks, both collected on vegetation by flagging and removed from humans. DNA of B. garinii was recorded most often; DNA of B. afzelii was less frequent; and the least number of positive samples was shown for B. miyamotoi. In the ticks collected on vegetation by flagging, DNA of B. garinii was found in 38.6%, B. afzelii in 9.9%, and B. miyamoboi in 3.9% of samples. In the ticks removed from people, number of positive samples was lesser; so, DNA of B. garinii was detected in 24.2%, B. afzelii in 6.9%, and B. miyamotoi in 5.6% of samples. Mixed infection with two Borrelia species was recorded, and DNA of B. mivamnotoi more often detected simultaneously with DNA of B. garinii.
Assuntos
Borrelia/crescimento & desenvolvimento , Borrelia/isolamento & purificação , Ixodes/microbiologia , Animais , Borrelia/classificação , Borrelia/genética , DNA Bacteriano/genética , Humanos , Reação em Cadeia da Polimerase/métodos , Sibéria , ÁrvoresRESUMO
48 full-length Borrelia garinii and Borrelia afzelii from West Siberia and Mongolia ospA gene nucleotide sequences was determined. Four groups of Borrelia garinii were revealed using the analysis of nucleotide sequences. The most variable ospA gene region was demonstrated to be included in region where the antigenic determinants of protein were encoded. High homology level was shown for nucleotide sequences corresponding to isolates of Borrelia afzelii.
Assuntos
Antígenos de Superfície/genética , Proteínas da Membrana Bacteriana Externa/genética , Vacinas Bacterianas/genética , Grupo Borrelia Burgdorferi/genética , Variação Genética , Lipoproteínas/genética , Homologia de Sequência do Ácido Nucleico , Mongólia , SibériaRESUMO
The 35 full-length Borrelia burgdorferi sensu lato complex a83/100 gene nucleotide sequences were determined. High level of homology was observed in the nucleotide sequences corresponding to the strains and isolates of Borrelia fzelii. The analysis of the nucleotide sequences revealed two groups of Borrelia garinii. The most variable p83/100 gene region containing species-typical insertions and deletions was demonstrated to be included into the region where the antigenic determinants of protein were encoded. According to the data obtained in this work, the modification of the P83/100 protein structure and immunological properties could be suggested to exist even within species. The results of this work could be used for receiving recombinant P83/100 proteins useful for diagnostic applications.
Assuntos
Antígenos de Bactérias/química , Antígenos de Bactérias/genética , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Grupo Borrelia Burgdorferi/classificação , Variação Genética , Sequência de Aminoácidos , Sequência de Bases , Grupo Borrelia Burgdorferi/genética , Grupo Borrelia Burgdorferi/imunologia , Genes Bacterianos , Dados de Sequência Molecular , Mutagênese Insercional , Filogenia , Análise de Sequência de DNA , Deleção de SequênciaRESUMO
Borrelia DNA was detected by polymerase chain reaction in the blood of patients who had suffered from tick suction. DNA was revealed in 28.7 of the blood samples from patients with the erythematous form of Ixodes tick-born borrelioses (ITBB) and in 14.3% of those diagnosed as having tick-borne encephalitis. Blood Borrelia DNA was not detected in patients with end-stage and chronic ITBB. Comparing the results of detection of DNA with those Borrelia protein antibodies has shown that the antibody titers detectable in the patients having Borrelia DNA are lower than those in the patients with the same form of ITBB and without DNA. The detection of DNA should be accomplished in the first 4 weeks after tick bite.
Assuntos
Grupo Borrelia Burgdorferi/isolamento & purificação , DNA Bacteriano/sangue , Doença de Lyme/diagnóstico , Doenças Transmitidas por Carrapatos/diagnóstico , Animais , Anticorpos Antibacterianos/sangue , Diagnóstico Precoce , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Ixodes/microbiologia , Reação em Cadeia da PolimeraseRESUMO
Detection of DNA of Borrelia burgdorferi sensu lato was performed by PCR in taiga ticks Ixodes persulcatus, in blood samples and skin bioptates of small forest mammals, and in blood and urine samples of humans after attaching of ticks events. In Novosibirsk region both in natural reservoir and in patients with Ixodes ticks-borne borreliosis DNA of Borrelia garinii and Borrelia afzelii are detected. DNA of these borrelia were detected in 8 from 72 of taiga ticks, in 36 from 298 of blood and skin samples of small forest mammals, and in 32 from 102 of human blood and urine samples. In all studied samples DNA of B. garinii from NT29 subgroup was predominated. Borrelia DNA in which sequence of intergenic spacer region was homologous to sequence Chy13p first detected in China has been detected in one blood sample from red-backed vole (Clethrionomys rutilus).
