Virulence Factors Present In Strains Of Porcine Enteropathogenic Escherichia Coli Isolated In Brasil [fatores De Virulencia Em Amostras De Escherichia Coli Enteropatogenicas Para Suinos Isoladas No Brasil]

[No abstract available]161213

Transposon Mutagenesis And Membrane Protein Studies In An Avian Colisepticaemic Escherichia Coli Strain

The pathogenicity, antibiotic resistance, plasmid DNA and membrane protein profiles of a colisepticaemic Escherichia coli strain (362) isolated from chickens was studied. It was verified that this strain harboured at least five plasmids. One 88.0 MD plasmid is non conjugative and is responsible for the production of colicin V and serum resistance. The 43.0 MD plasmid is responsible for resistance to bactericidal activity of serum and ampicilin. Curing of these plasmids did not decrease the pathogenicity of the strain. Transposon mutagenesis (Tnpho A) of strain 362 yielded a non-pathogenic derivative strain which had lost a 40.7 kD membrane protein, which is not correlated to the aerobactin and enterochelin systems. We suggest that this protein subunit is involved in the pathogenicity process of avian septicaemic E. coli strains.17191

Bluetongue Virus: Production And Study Of Viral Antigen For Serological Diagnosis

A soluble antigen, produced from the culture supernatant of VERO cells infected with bluetongue virus serotype 4 (BTV-S4) and concentrated by sequential ultrafiltration with membranes with cut-off values 103 and 25 × 103 NMWP, showed complete identity to standard antigens when compared by agar gel immunodiffusion (AGID) and SDS-PAGE profiles, revealing that the main protein component responsible for the AGID reaction has a molecular weight of about 60 kDa corresponding probably to the NS1 protein. © 1993.4402/03/15281286Adams, Gogolewski, Barbet, Cheevers, Identification of caprine arthritisencephalitis retrovirus proteins in immunodiffusion precipitin line (1985) J. Gen. Virol., 66, pp. 1139-1143Campbell, Grubman, Current knowledge on the biochemistry and immunology of bluetongue (1985) Prog. Vet. Microbiol. Immunol., 1, pp. 58-79Eaton, Hyatt, White, Localization of the nonstructural protein NS1 in bluetongue virus-infected cells and its presence in virus particles (1988) Virology, 163, pp. 527-537Hubschle, Yang, Immunodiffusion studies with bluetongue virus using an isolated core protein (1983) Proc. Am. Assoc. Vet. Lab. Diagn., 26, pp. 725-730Huismans, Protein synthesis in bluetongue virus-infected cells (1979) Virology, 92, pp. 385-396Huismans, Cloete, A comparison of different cloned bluetongue virus genome segments as probes for the detection of virus-specified RNA (1987) Virology, 158, pp. 373-380Huismans, Els, Characterization of the tubules associated with the replication of three different orbiviruses (1979) Virology, 92, pp. 397-406Huismans, Bremer, Barber, The nucleic acid and proteins of epizootic haemorrhagic disease virus (1979) J. Vet. Res., 46, pp. 51-58Jochim, Chow, Immunodiffusion of bluetongue virus (1969) Am. J. Vet. Res., 30, pp. 33-41Jochim, Improvement of the AGP test for bluetongue (1976) Proc. Am. Assoc. Vet. Lab. Diagn., 19, pp. 361-376Jochim, Pearson, Protocol for the immunodiffusion test for bluetongue (1979) Proc. Am. Assoc. Vet. Lab. Diagn., 22, pp. 463-471Jochim, An overview of diagnostics for bluetongue (1985) B. Jochim, Bluetongue and Related Orbiviruses, pp. 423-433. , Alan R. Liss, New YorkKlontz, Svehag, Gorhan, A study by the agar diffusion technique of precipitating antibody directed against blue tongue virus and its relation to hemotypic neutralizing antibody (1962) Archiv f�r die gesamte Virusforschung, 2, pp. 259-272Knudson, Shope, Overview of the orbiviruses (1985) B. Jochim. Bluetongue and Related Orbiviruses, pp. 255-266. , Alan R. Liss, New YorkLaemmli, Cleavage of structural proteins during the assembly of the head of bacteriophage T4 (1970) Nature, 227, pp. 680-685Matthews, Classification and nomenclature of viruses (1982) Fourth Rep. Int. Com. Tax. Vir. Intervirol., 17, pp. 1-199Mechan, Dean, Jochim, Correlation of serotype specificity and protein structure of the five U.S. serotypes of bluetongue virus (1986) J. Gen. Virol., 67, pp. 2617-2624Ranger, Brown, Bluetongue/epizootic haemorrhagic disease agar gel immunodiffusion (AGID) antigen (1985) NVSL Diagnostic Reagents Production Guide No. R-63/94, pp. 1-4Verwoerd, Els, De, Huismans, Structure of the bluetongue virus capsid (1972) J. Virol., 10, pp. 783-794Verwoerd, Huismans, Studies on the in vitro and the in vivo transcription of the bluetongue virus genome (1972) Onderstepoort J. Vet. Res., 39, pp. 185-192Urakawa, Roy, Bluetongue virus tubules made in insect cells by recombinant baculoviruses: expression on the NS1 gene of bluetongue virus serotype 10 (1988) J. Virol., 62, pp. 3919-3927Wang, Luedke, Chow, Soluble antigen of bluetongue virus (1972) Inf. Immunol., 5, pp. 467-47

Rotavirus Excretion In Naturally Infected Pigs With And Without Diarrhoea

Seven hundred and fifty faecal samples from piglet ranging from 1 to 60 days old were studied for the presence of group A rotavirus by polyacrylamide gel electrophoresis (PAGE) and by enzyme immunoassay (EIA). From 451 diarrhoeic pigs, 117 (25.94%) were positive for rotavirus and only 45 (15.05%) of 299 pigs without diarrhoea excreted the virus (P < 0.005). When these animals were separated into four age groups with regard to the presence or absence of diarrhoea, it was observed that the excretion of rotavirus was associated with diarrhoea in piglets, both before and after weaning. © 1993.3701/02/15187190Alpers, Sandres, Hampson, Rotavirus excretion by village pigs in Papua New Guinea (1991) Austr. Vet. J., 68, pp. 65-67Benfield, Stotz, Moore, McAdhragh, Shedding of rotavirus in faeces of sows before and after farrowing (1982) J. Clin. Microbiol., 161, pp. 186-190Bohl, Rotaviral diarrhea in pigs: brief review (1979) J. Am. Vet. Med., 174, pp. 613-615Fu, Hampson, Group A rotavirus excretion patterns in naturally infected pigs (1987) Res. Vet. Sci., 43, pp. 297-300Herring, Inglis, Ojeh, Snodgrass, Menzies, Rapid diagnosis of rotavirus infection by directed detection of viral nucleic acid in silver stained polyacrylamide gels (1982) J. Clin. Microbiol., 16, pp. 473-477Laemmli, Cleavage of structural proteins during the assembly of the head of bacteriophage T4 (1970) Nature (London), 227, pp. 680-685Pereira, Azevedo, Leite, Andrade, de Castro, A combined enzyme immunoassay for rotavirus and adenovirus (EIARA) (1985) Journal of Virological Methods, 10, pp. 21-28Tzipori, Chandler, Smith, Makin, Hennessy, Factors contributing to postweaning diarrhea in a large intensive piggery (1980) Aust. Vet. J., 56, pp. 274-27

Studies Of The Genetic Expression Of 31a Fimbriae By Two Bovine Septicaemic Escherichia Coli Strains

Using transposon tnphoA, classical bacterial genetic assays, SDS-PAGE and Western blot of superficial proteins, we have studied the expression of the 31A fimbriae of two bovine wild-type septicaemic Escherichia coli strains. The genes responsible for encoding colonization factor 31A were located in both the plasmid (strain BZ2468) and the chromosome (strain BZ43). The results obtained using HeLa cell cultures led us to believe that the BZ43 strain could have another colonization factor besides 31 A, since one mutant, which did not express fimbriae, was still able to adhere to HeLa cells.17436537

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. Microbiol, 21, pp. 189-194Dho, M., Lafont, J.P., Adhesive properties and iron uptake ability in Escherichia coli lethal and non-lethal for chicks (1984) Avian Dis, 28, pp. 1016-1025Dho-Moulin, M., Fairbrother, J.M., Avian pathogenic Escherichia coli (APEC) (1999) Vet. Res, 30, pp. 299-316Donnenberg, M.S., Kaper, J.B., Enteropathogenic Escherichia coli (1992) Infect. Immun, 60, pp. 3953-3961Dozois, C.M., Dho-Moulin, M., Bree, A., Fairbrother, J.M., Desautels, C., Curtis III, R., Relationship between the Tsh autotransporter and pathogenicity of avian Escherichia coli and localization and analysis of the genetic region (2000) Infect. Immun, 68, pp. 4145-4154Fantinatti, F., Silveira, W.D., Castro, A.F.P., Characteristics associated with pathogenicity of avian septicaemic Escherichia coli strains (1994) Vet. 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. Microbiol, 30, pp. 1189-1193Macrina, F.L., Kopecko, D.J., Jones, K.R., Ayers, D.J., McCowen, S.M., A multiple plasmid-containing Escherichia coli strain: Convenient source of size reference plasmid molecules (1978) Plasmid, 1, pp. 417-420Marc, D., Dho-Moulin, M., Analysis of the fim cluster of an avian O2 strain of Escherichia coli: Serogroup-specific sites within fimA and nucleotide sequence of fimI (1996) J. Med. Microbiol, 44, pp. 444-452Maurer, J.J., Brown, T.P., Steffens, W.L., Thayer, S.G., The occurrence of ambient temperature-regulated adhesins, curli, and temperature-sensitive hemagglutinin Tsh among avian Escherichia coli (1998) Avian Dis, 42, pp. 106-118Nataro, J.P., Kaper, J.B., Diarrheagenic Escherichia coli (1998) Clin. Microbiol. Rev, 11, pp. 142-201Naveh, M.W., Zusman, T., Skutelsky, E., Ron, E.Z., Adherence pili in avian strains of Escherichia coli: Effect on pathogenicity (1984) Avian Dis, 28, pp. 651-661Nicholls, L., Grant, T.H., Robins-Browne, R.M., Identification of a novel genetic locus that is required for in vitro adhesion of a clinical isolate of enterohaemorrhagic Escherichia coli to epithelial cells (2000) Mol. Microbiol, 35, pp. 275-288Parsot, C., Shigella spp. and enteroinvasive Escherichia coli pathogenicity factors (2005) FEMS Microbiol. Lett, 252, pp. 11-18Pourbakhsh, S.A., Dho-Moulin, M., Brée, A., Desautels, C., Martineau-Doizé, B., Fairbrother, J.M., Localization of the in vivo expression of P and F1 fimbriae in chickens experimentally inoculated with pathogenic Escherichia coli (1997) Microb. Pathog, 22, pp. 331-341Prasadarao, N.V., Wass, C.A., Stins, M.F., Shimada, H., Kim, K.S., Outer membrane protein A-prometed actin condensation of brain microvascular endothelial cells is required for Escherichia coli invasion (1999) Infect. Immun, 67, pp. 5775-5783Provence, D.L., Curtiss III, R., Isolation and characterization of a gene involved in hemagglutination by an avian pathogenic Escherichia coli strain (1994) Infect. Immun, 62, pp. 1369-1380Rasmussen, H.N., Rasmussen, O.F., Andersen, J.K., Olsen, J.E., Specific detection of pathogenic Yersinia enterocolitica by two-step PCR using hot-start and DMSO (1994) Mol. Cell. Probes, 8, pp. 99-108Sambrook, J., Fritsch, E.F., Maniatis, T., (1989) Molecular Cloning: A Laboratory Manual, , 2nd edn. Cold Spring Harbor Laboratory Press: New YorkSansonetti, P.J., Ryter, A., Clerc, P., Maurelli, A.T., Mounier, J., Multiplication of Shigella flexneri within HeLa cells: Lysis of the phagocytic vacuole and plasmid-mediated contact hemolysis (1986) Infect. Immun, 51, pp. 461-469Scaletsky, I.C.A., Silva, M.L.M., Trabulsi, L.R., Distinctive patterns of adherence of enteropathogenic Escherichia coli to HeLa cells (1984) Infect. Immun, 45, pp. 534-536Schmidt, H., Zhang, W.L., Hemmrich, U., Jelacic, S., Brunder, W., Tarr, P.I., Dobrindt, U., Karch, H., Identification and characterization of a novel genomic island integrated at selC in locus of enterocyte effacement-negative, Shiga toxin-producing Escherichia coli (2000) Infect. Immun, 69, pp. 6863-6873Silveira, W.D., Ferreira, A., Brocchi, M., Hollanda, L.M., Pestana de Castro, A.F., Yamada, A., Lancellotti, M., Biological characteristics and pathogenicity of avian Escherichia coli strains (2002) Vet. 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Genes Coding For Shiga-like Toxins In Bovine Verotoxin-producing Escherichia Coli (vtec) Strains Belonging To Different O:k:h Serotypes

Forty-six verotoxin-producing Escherichia coli (VTEC) strains isolated from diarrhoeic and healthy calves in Spain were examined for DNA sequences homologous to genes for verotoxins (VT1 and VT2) and enterotoxins (LT-I, LT-II, STaH, STaP and STb). Hybridisation showed that 26 (57%) of VTEC strains carried VT1 genes, 13 (28%) possessed VT2 genes, and 7 (15%) carried both VT1 and VT2 genes. No VTEC strains hybridised with DNA probes for enterotoxins. A correlation was found between the serotype and type of VT produced. Thus, all strains of serotypes O26:K-:H11 (13 strains), O103:K-:H2 (3 strains) and O128:K?:H- (4 strains) hybridised with the VT1 probe only, whereas all strains of serotypes O4:K-:H4 (3 strains) and O113:K-:H21 (4 strains) were positive with the VT2 probe only. By contrast, O81:K?:H28 (2 strains) and O157:K-:H- (2 strains) strains hybridised with both VT1 and VT2 probes. One strain of serotype O157:K-:H7 was VT2 positive. © 1994.422-3105110Barret, Kaper, Jerse, Wachsmuth, Virulence factors in Shiga-like toxin-producing Escherichia coli isolated from humans and cattle (1992) J. Infect. Dis., 165, pp. 979-980Blanco, González, García, Blanco, Regueiro, Bernárdez, Production of toxins by Escherichia coli strains isolated from calves with diarrhoea in Galicia (North-western Spain) (1988) Vet. Microbiol., 18, pp. 297-311Blanco, Blanco, Blanco, Ramos, Enterotoxigenic, verotoxigenic and necrotoxigenic Escherichia coli isolated from cattle in Spain (1993) Am. J. Vet. Res., 54, pp. 1446-1451Burnens, Boss, Ørskov, Ørskov, Schaad, Müller, Heinzle, Nicolet, Occurrence and phenotypic properties of verotoxin producing Escherichia coli in sporadic cases of gastroenteritis (1992) Eur. J. Clin. Microbiol. Infect. 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IDENTIFICAÇÃO DOS GENES QUE CODIFICAM PARA A ENTEROTOXINA TERMOLÁBIL LT-II EM AMOSTRAS DE ESCHERICHIA COLI ISOLADAS DE BEZERROS COM DIARRÉIA NA REGIÃO DE JABOTICABAL, SP, BRASIL IDENTIFICATION OF THE GENES THAT ENCODE FOR THE LT-II THERMOLABILE ENTEROTOXIN AMONG STRAINS OF ESCHERICHIA COLI ISOLATED FROM CALVES WITH DIARRHEA IN THE REGION OF JABOTICABAL, SP, BRAZIL)

Examinando 52 espécimes fecais de bezerros com diarréia de fazendas da região de Jaboticabal, SP, Brasil, uma das amostras de Escherichia coli isoladas, quando analisada pela Reação em Cadeia da Polimerase (PCR), demonstrou a presença dos genes que codificam para a enterotoxina termolábil do tipo LT-II. Este é o primeiro relato de amostra de E. coli enterotoxigênica, isolada de bezerros com diarréia no Brasil, contendo os genes para codificação da enterotoxina LT-II. Encontram-se apenas citações de isolamento no Brasil de amostras de E. coli LT-II+ de alimentos de origem animal.<br>Examining 52 samples of stools from calves with diarrhea from the region of Jaboticabal, SP, Brazil , one of the strains of Escherichia coli isolated, as examined by the Polymerase Chain Reaction (PCR), harbored the genes encoding for the LT-II thermolabile enterotoxin. This is the first report on the isolation of enterotoxigenic E. coli from calves with diarrhea in Brazil harboring the genes for LT-II enterotoxin. In our country, strains of LT-II + E. coli have been reported to occur only in foods of animal origin