Multidrug-resistant Pseudomonas Aeruginosa: An Endemic Problem In Brazil [pseudomonas Aeruginosa Multirresistente: Um Problema Endêmico No Brasil]

Global reports have documented the endemicity of multidrug-resistant (MDR) Pseudomonas aeruginosa associated with high levels of morbidity/mortality. In Brazil, outbreaks of MDR P. aeruginosa have been related to clonal dissemination. Currently, therapeutic options for the treatment of these infections are restricted to carbapenemic antibiotics (i.e., imipenem [IPM]). Thus, carbapenem resistance is a public health issue, since carbapenems are considered the last resort to nosocomial infections caused by MDR Gram-negative bacteria. In Brazil, the main mechanisms associated with MDR P. aeruginosa phenotypes are metallo-betalactamase (MBL) production (SPM-1 enzyme), presence of 16S rRNA methylase RmtD, loss of OprD porin, and overexpression of efflux pumps, which may explain the high level of carbapenem and aminoglycoside resistance. Accordingly, the emergence and dissemination of MDR strains is worrisome. 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Improvement of the indirect hemagglutination test for the detection of antibodies to Streptococcus pyogenes

An indirect hemagglutination test for a seroepidemiological survey of Streptococcus pyogenes infection was standardized. This is an improved modification of the indirect hemagglutination test which utilizes an unstable reagent prepared with fresh blood cells. Two types of bacterial antigens represented by extracellular products and purified streptolysin O were assayed, but only the former antigen gave good results. Pretreatment of the bacterial antigen with 0.15 M NaOH and neutralization to pH 5.5, as well as postfixation of sensitized red cells with 0.1% glutaraldehyde at 56oC for 30 min were found to be essential to give long stability to the reagent in liquid suspension, at least 9 months at 4oC. A total of 564 serum samples with high, moderate and low anti-streptolysin O antibodies as determined by the neutralization assay were studied by the indirect hemagglutination test using the new reagent. The sensitivity, specificity, efficiency, positive predictive value and negative predictive value of the test in relation to the neutralization assay were 0.950, 0.975, 0.963, 0.973, and 0.955, respectively. The kappa agreement index between the two techniques was high (0.926) and ranked as "almost perfect". Antibody levels detected by both techniques also presented a high positive correlation (rs = 0.726). Five reagent batches successively produced proved to be reproducible. Thus, the improved indirect hemagglutination test seems to be useful for public health laboratories

Carbapenem-resistant Acinetobacter baumannii outbreak at university hospital Caracterização de cepas de Acinetobacter baumannii durante um surto de infecção hospitalar

Nineteen clonally related imipenem-resistant Acinetobacter baumannii isolates were recovered from eight intensive care unit patients. All isolates harboured blaOXA-51-like &#946;-lactamase genes and showed the absence of 22 kDa fraction in outer membrane porin profile analysis. It suggests a combination of two mechanisms as responsible for carbapenemresistant phenotypes.<br>Foram isoladas 19 cepas monoclonais de 8 pacientes da unidade de terapia intensiva, resistentes aos carbapenêmicos. Todas as cepas apresentaram o gene blaOXA-51-like e por análise do perfil de proteínas de membrana notou-se ausência da fração de 22 kDa, sugerindo a combinação de dois mecanismos de resistência aos carbapenêmicos

Estafilococos resistentes à oxacilina isolados em casos de mastite subclínica em ovinos

Bactérias do gênero Staphylococcus estão entre os principais agentes causadores da mastite ovina. Um dos maiores entraves ao tratamento dos animais doentes são cepas resistentes aos antimicrobianos empregados. A pesquisa do gene mecA nos estafilococos é um instrumento auxiliar para a determinação de aspectos epidemiológicos da doença. Este trabalho teve por objetivo investigar a resistência à oxacilina em estafilococos coagulase-negativos isolados no leite de ovelhas com mastite subclínica. Foram analisadas 448 amostras de leite de dois rebanhos. Os micro-organismos isolados foram submetidos previamente a testes de sensibilidade a antibióticos in vitro a partir da técnica de difusão em disco. Naqueles resistentes à oxacilina nestes testes efetuou-se a pesquisa do gene mecA, com a extração do DNA cromossômico por meio da técnica de extração fenol-clorofórmio. Os estafilococos coagulase-negativos apresentaram resistência à oxacilina e a presença do gene mecA foi detectada em quatro isolados, que também apresentaram características de multirresistência. Tais achados reforçam a importância deste grupo de micro-organismos na etiologia da mastite subclínica em ovinos e abre perspectivas para futuras pesquisas para a investigação da epidemiologia da doença

Molecular characterization of van genes found in vancomycin-resistant Enterococcus spp. isolated from Hospital das Clínicas, FMUSP, São Paulo, Brazil

Vancomycin-resistant enterococci strains (VRE) is an important pathogen related with hospital infections in many countries, presenting limited or no therapeutic options for treating serious infections. VRE has presented some different genotypes been VanA and VanB considered to be the most important in hospital environments. In the present study the authors investigated the prevalence of van genes (A, B an C) among clinical isolates of VRE in a five month period at a large tertiary hospital in Sao Paulo, Brazil. The results showed the presence of vanA, but not vanB or vanC in all 43 strains of E. faecalis and five E. faecium studied. The results bring an important issue, due to the possibility of resistance spread of vanA genes, to be monitored and solved by the hospital infection control team and the microbiology and molecular biology laboratories at tertiary Hospitals

Complete Nucleotide Sequences Of Two Blakpc-2-bearing Incn Plasmids Isolated From Sequence Type 442 Klebsiella Pneumoniae Clinical Strains Four Years Apart

We sequenced the oldest blaKPC-2-bearing plasmid isolated in Brazil and another plasmid also carried by a Klebsiella pneumoniae strain of sequence type 442 (ST442), isolated 52 months later. Both plasmids present an IncN backbone and few acquired regions. Because the 2005 plasmid presented deletions and a truncated gene within Tn4401b compared to the 2009 plasmid, we can thus infer that IncN blaKPC-2-bearing plasmids pFCF1305 and pFCF3SP had a common ancestor circulating in Brazil prior to May 2005.© 2014, American Society for Microbiology. All Rights Reserved.58529582960Monteiro, J., Santos, A.F., Asensi, M.D., Peirano, G., Gales, A.C., First report of kpc-2-producing klebsiella pneumoniae strains in brazil (2009) Antimicrob Agents Chemother, 53, pp. 333-334. , http://dx.doi.org/10.1128/AAC.00736-08Pavez, M., Mamizuka, E.M., Lincopan, N., Early dissemination of kpc-2-producing klebsiella pneumoniae strains in brazil (2009) Antimicrob Agents Chemother, 53, p. 2702. , http://dx.doi.org/10.1128/AAC.00089-09Sambrook, J., Russell, D.W., (2001) Molecular Cloning: A Laboratory Manual, , 3rd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYChevreux, B., Wetter, T., Suhai, S., Genome sequence assembly using trace signals and additional sequence information (1999) Computer Science And Biology: Proceedings Of The German Conference On Bioinformatics (GCB, 99, pp. 45-56Kearse, M., Moir, R., Wilson, A., Stones-Havas, S., Cheung, M., Sturrock, S., Buxton, S., Drummond, A., Geneious basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data (2012) Bioinformatics, 28, pp. 1647-1649. , http://dx.doi.org/10.1093/bioinformatics/bts199Altschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.J., Basic local alignment search tool (1990) J. Mol. Biol, 215, pp. 403-410Aziz, R.K., Bartels, D., Best, A.A., DeJongh, M., Disz, T., Edwards, R.A., Formsma, K., Zagnitko, O., The rast server: Rapid annotations using subsystems technology (2008) BMC Genomics, 9, p. 75. , http://dx.doi.org/10.1186/1471-2164-9-75Okonechnikov, K., Golosova, O., Fursov, M., Unipro ugene: A unified bioinformatics toolkit (2012) Bioinformatics, 28, pp. 1166-1167. , http://dx.doi.org/10.1093/bioinformatics/bts091Siguier, P., Perochon, J., Lestrade, L., Mahillon, J., Chandler, M., Isfinder: The reference centre for bacterial insertion sequences (2006) Nucleic Acids Res, 34, pp. D32-D36. , http://dx.doi.org/10.1093/nar/gkj014Naas, T., Cuzon, G., Villegas, M.-V., Lartigue, M.-F., Quinn, J.P., Nordmann, P., Genetic structures at the origin of acquisition of the beta-lactamase bla kpc gene (2008) Antimicrob Agents Chemother, 52, pp. 1257-1263. , http://dx.doi.org/10.1128/AAC.01451-07Papagiannitsis, C.C., Miriagou, V., Giakkoupi, P., Tzouvelekis, L.S., Vatopoulos, A.C., Characterization of pkp1433, a novel kpc-2-encoding plasmid from klebsiella pneumoniae sequence type 340 (2013) Antimicrob Agents Chemother, 57, pp. 3427-3429. , http://dx.doi.org/10.1128/AAC.00054-1