Dna Sequencing Of A Pathogenicity-related Plasmid Of An Avian Septicemic Escherichia Coli Strain.

A 43-MDa conjugative plasmid isolated from an avian septicemic Escherichia coli (APEC) strain possessing genes related to the adhesion and invasion capacities of in vitro-cultured cells was sequenced. The results demonstrated that the 43-MDa plasmid harbors bacterial pathogenicity-related sequences which probably allow the wild-type pathogenic strain to adhere to and invade tissues and to cause septicemia in poultry. The existence of homology sequences to sequences belonging to other human pathogenic Enterobacteriaceae like Escherichia coli O157:H7, Shigella and Salmonella was also observed. The presence of these sequences in this plasmid could indicate that there is horizontal genetic transfer between bacterial strains isolated from different host species. In conclusion, the present study suggests that APEC strains harbor high-molecular weight plasmids that present pathogenicity-related sequences and that these are probably responsible for the pathogenicity exhibited by these strains. The presence of human pathogenicity-associated sequences in APEC conjugative plasmids suggests that these strains could represent a zoonotic risk.6223123

Molecular Epidemiology Of Shigella Spp Strains Isolated In Two Different Metropolitam Areas Of Southeast Brazil [epidemiologia Molecular De Linhagens De Shigella Spp Isoladas Em Diferentes Areas Metropolitanas Do Sudeste Do]

Shigella spp., the human pathogen responsible for shigellosis, is highly infectious even at low levels. The incidence rate of shigellosis varies with geographical distribution, location human development index, and age groups, being higher among children aged under 5 years. In Brazil, a few works indicate that shigellosis cases are underestimated, with S. flexneri and S. sonnei strains being the major agents responsible for the shigellosis cases. The present study used pulsed field gel electrophoresis (PFGE) to investigate the molecular epidemiology of 119 strains of S. sonnei and S. flexneri isolated from shigellosis cases that occurred in the metropolitan areas of Ribeirão Preto and Campinas Cities, São Paulo Sate, southeast Brazil. The results indicated (i) the existence of just a few strain clusters for both species, but with genotype variability with either a high speed of genetic change or constant introduction of several genotypes, considering the intense migration to these two metropolitan areas, and (ii) the prevalence of specific genotypes in each geographical area, which suggests the successful adaptation of some genotypes to the local environmental conditions. Our results indicate the need of more efficacious sanitary barriers to prevent Shigella spp. outbreaks and epidemics.403685692Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Smith, J.A., Seidman, J.G., Struhl, K., (1998) Current Protocols in Molecular Biology, p. 1. , New York: Green Publishing Associates1.6Brenner, D.J., Fanning, G.R., Skeman, F.J., Falkow, S., Polynucleotide sequence divergence among strains of Escherichia coli and closely related organisms (1972) J. Bacteriol., 109, pp. 953-965Delappe, N., O'Halloran, F., Fanning, S., Corbett-Feeney, G., Cheasty, T., Cormican, M., Antimicrobial resistance and genetic diversity of Shigella sonnei isolates from western Ireland, an area of low incidence of infection (2003) J. Clin. Microbiol., 41, pp. 1919-1924Diniz-Santos, D.R., Santana, J.S., Barreto, J.R., Andrade, M.G., Silva, L.R., Epidemiological and microbiological aspects of acute bacterial diarrhea in children from Salvador, Bahia, Brazil (2005) Braz. J. Infect. Dis., 9, pp. 77-83Ferreccio, C., Prado, V., Ojeda, A., Cayyazo, M., Abrego, P., Guers, L., Levine, M.M., Epidemiologic patterns of acute diarrhea and endemic Shigella infections in a poor periurban setting in Santiago (1991) Chile. Am. J. Epidemiol., 134, pp. 614-627Kotloff, K.L., Winickoff, J.P., Ivanoff, B., Clemens, J.D., Swerdlow, D.L., Sansonetti, P.J., Adak, G.K., Levine, M.M., Global burden of Shigella infections: Implications for vaccine development and implementation of control strategies (1999) Bull. World Health Organ, 77, pp. 651-666Levine, M.M., Immunization against bacterial diseases of the intestine (2000) J. Pediatr. Gastroenterol. Nutr., 31, pp. 336-355Levine, M.M., Kotloff, K.L., Barry, E.M., Pasetti, M.F., Sztein, M.B., Clinical trials of Shigella vaccines: Two steps forward and one step back on a long, hard road (2007) Nat. Rev. Microbiol., 5, pp. 540-553Melo, M.C.N., Taddei, J.A.A.C., Diniz-Santos, D.R., May, D.S., Carneiro, N.B., Silva, L.R., Incidence of diarrhea: Poor parental recall ability (2007) Braz. J. Infect. Dis., 11, pp. 571-579Ochman, H., Whittam, T.S., Caugant, D.A., Selander, R.K., Enzyme polymorphism and genetic population structure in Escherichia coli and Shigella (1983) J. Gen. 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Infect., 136, pp. 577-603. , Part IIRégua-Mangia, A.H., Gomes, T.A.T., Andrade, J.R.C., Vieira, M.A.M., Gonzalez, A.G.M., Zahner, V., Irinos, K., Teixeira, L.M., Genetic Analysis of Escherichia coli strains carrying enteropathogenic Escherichia coli (EPEC) markers, isolated from children in Rio de Janeiro city (2003) Brazil. Braz. J. Microbiol., 34, pp. 38-41Sambrook, J., Fritsch, E.F., Maniatis, T., (1989) Molecular cloning: A laboratory manual, , 2nd edn. New York: Cold Spring Harbor Laboratory PressSmall, P., Blankenhorn, D., Welty, D., Acid and base resistance in Escherichia coli and Shigella flexneri: Role of rpoS and growth pH (1994) J. Bacteriol., 176, pp. 1729-1737Straden, A., Frei, R., Widmer, A.F., Molecular typing of methicillin-resistant Staphylococcus aureus: Can PCR replace pulsed- field gel electrophoresis? (2003) J. Clin. 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Study Of The Adhesion And Invasion Capacities Of An Invasive Avian Pathogenic Escherichia Coli Strain (apec) To In Vitro Cultivated Hep-2 Cells

An avian pathogenic Escherichia coli strain (APEC), designated strain sep17, was isolated from the liver of a chicken suffering from septicaemia. Its biological characteristics, including antimicrobial drug resistances, colicin production, plasmid profile, adhesion and invasion capacities to in vitro cultivated HEp-2 cells, and PCR detection of DNA sequences related to pathogenicity genes (fimA, tsh, papA, crl, csgA, afa, sfa, eae, lpfA O157/OI-141, lpfAO157/OI-154, toxB, iha, ial, efa, inv, invA/invE and ibeA) were determined. This strain (Lac+, Tc R, fimA, csgA, crl, lpfAO157/OI-154) harbored a 70 MDa plasmid and was able to adhere to and invade in vitro cultured HEp-2 cells. Transference of this 70 MDa plasmid by conjugation rendered a non-pathogenic recipient strain HB101 (Lac-, SmR, csgA) capable of adhering to and invading the same type of cell. Scanning electron microscopy and light microscopy confirmed the adhesion capacity of the wild and the transconjugant strains, while the in vitro invasion technique and light microscopy confirmed the invasion capacity of these strains. The FAS technique, which is used to visualize actin accumulation on cells where the adhesion process occurs, was negative for all those strains. None of the genes detected in strain sep17 were transferred by conjugation, which indicates that they are chromosomally located and are not related to the adhesion and invasion processes. The presence and absence of pathogenicity-related genes are discussed.241110Amabile de Campos, T.A., Stehling, E.G., Ferreira, A., Pestana de Castro, A.F., Brocchi, M., Silveira, W.D., Adhesion properties, fimbrial expression and PCR detection of adhesin-related genes of avian Escherichia coli strains (2005) Vet. Microbiol, 106, pp. 275-285Azevedo, J.L., Da Costa, S.O.P., (1973) Exercícios Práticos de Genética, , Companhia Editora Nacional, Editora da Universidade de São Paulo, São Paulo: SPBenjamin, P., Federman, M., Wanke, C.A., Characterization of an invasive phenotype associated with enteroaggregative Escherichia coli (1995) Infect. Immun, 63, pp. 3417-3421Blanco, M., Blanco, J.E., Alonso, M.P., Mora, A., Balsalobre, C., Munoa, F., Juárez, A., Blanco, J., Detection of pap, sfa, and afa adhesin-encoding operons in uropathogenic Escherichia coli strains: Relationship with expression of adhesins and production of toxins (1997) Res. Microbiol, 148, pp. 745-755Boyer, H.W., Roulland-Dussoix, D., A complementation analysis of the restriction and modification of DNA in Escherichia coli (1969) J. Mol. Biol, 41, pp. 459-472Chulasiri, M., Suthienkul, O., Antimicrobial resistance of Escherichia coli isolated from chickens (1989) Vet. 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Microbiol, 41, pp. 75-86Gannon, V.P.J., Rashed, M., King, R.K., Thomas, E.J.G., Detection and characterization of the eae gene of Shiga-like toxin-producing Escherichia coli using polymerase chain reaction (1993) J. Clin. Microbiol, 31, pp. 1268-1274Gelderblom, H.R., Hausmann, E.H., Ozel, M., Pauli, G., Koch, M.A., Fine structure of human immunodeficiency virus (HIV) and immunolocalization of structural proteins (1987) Virology, 156, pp. 171-176Germon, P., Chen, Y.H., He, L., Blanco, J.E., Brée, A., Schouler, C., Huang, S.H., Moulin-Schouleur, M., ibeA, a virulence factor of avian pathogenic Escherichia coli (2005) Microbiology, 151, pp. 1179-1186Gophna, U., Oelschlaeger, T.A., Hacker, J., Ron, E.Z., Yersinia HPI in septicemic Escherichia coli strains isolated from diverse hosts (2001) FEMS Microbiol. Lett, 196, pp. 57-60Gross, W.B., Colibacillosis (1991) Disease of Poultry, pp. 138-144. , Calnek BW, Barnes HJ, Beard CW, Reid WM, Yoder JHW, eds, pp, Iowa State University Press: Ames, IowaKim, B.Y., Kang, J., Kim, K.S., Invasion processes of pathogenic Escherichia coli (2005) Int. J. Med. Microbiol, 295, pp. 463-470Knutton, S., Baldwin, T., Williams, P.H., McNeish, A.S., Actin accumulation at sites of bacterial adhesion to tissue culture cells: Basis of a new diagnostic test for enteropathogenic and enterohemorrhagic Escherichia coli (1989) Infect. Immun, 57, pp. 1290-1298Knutton, S., Phillips, A.D., Smith, H.R., Gross, R.J., Shaw, R., Watson, P., Price, E., Screening for enteropathogenic Escherichia coli in infants with diarrhea by the fluorescent-actin staining test (1991) Infect. Immun, 59, pp. 365-371La Ragione, R.M., Cooley, W.A., Woodward, M.J., The role of fimbriae and flagella in the adherence of avian strains of Escherichia coli O78:K80 to tissue culture cells and tracheal and gut explants (2000) J. Med. Microbiol, 49, pp. 327-338La Ragione, R.M., Sayers, A.R., Woodward, M.J., The role of fimbriae and flagella in the colonization, invasion and persistence of Escherichia coli O78:K80 in the day-old-chick model (2000) Epidemiol. Infect, 124, pp. 351-363Le Bouguenec, C., Archambaud, M., Labigne, A., Rapid and specific detection of the pap, afa, and sfa adhesin-encoding operons in uropathogenic Escherichia coli strains by polymerase chain reaction (1992) J. Clin. 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Molecular genotyping and epidemiology of Mycobacterium tuberculosis isolates obtained from inmates of correctional institutions of Campinas, Southeast Brazil

The objective of this study was to investigate the possible transmission of tuberculosis among 39 inmates with positive Mycobacterium tuberculosis smears in four correctional institutions located in Campinas City, SP, Brazil over a 19-month period. Fifty-one M. tuberculosis isolates from these inmates were characterized according to the number of IS6110 insertion elements present in their genomic DNA. The number of insertion elements in M. tuberculosis isolates varied from two to twelve. The dendrogram of similarity resulted in the grouping the isolates in six main clusters. These results, associated to epidemiological data, suggested the transmission of tuberculosis among inmates of the same and different institutions inmates. Univariate analysis of epidemiological data (total delay for beginning of treatment, previous treatment, and HIV status) and clustering occurrence showed that only "previous treatment" (OR = 7.65, p = 0.032) was associated with the possible transmission of tuberculosis in the studied prisons