Determination of flagellar types by PCR-RFLP analysis of enteropathogenic Escherichia coli (EPEC) and Shiga toxin-producing E. coli (STEC) strains isolated from animals in São Paulo, Brazil

2012 ◽  
Vol 92 (1) ◽  
pp. 18-23 ◽  
Author(s):  
Claudia de Oliveira Ayala ◽  
Ana Carolina Ramos Moreno ◽  
Marina Baquerizo Martinez ◽  
Ylanna Kelner Burgos ◽  
Antonio Fernando Pestana de Castro ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Rflp Analysis ◽  
Sao Paulo ◽  
São Paulo ◽  
Enteropathogenic Escherichia Coli ◽  
E Coli ◽  
Pcr Rflp

scholarly journals Molecular Subtyping and Genetic Analysis of the Enterohemolysin Gene (ehxA) from Shiga Toxin-Producing Escherichia coli and Atypical Enteropathogenic E. coli

2007 ◽  
Vol 73 (20) ◽  
pp. 6360-6369 ◽  
Author(s):  
Adrian L. Cookson ◽  
Jenny Bennett ◽  
Fiona Thomson-Carter ◽  
Graeme T. Attwood
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Diarrheal Disease ◽  
Rflp Analysis ◽  
Subtype C ◽  
E Coli ◽  
Pcr Rflp ◽  
Subtype A ◽  
Insight Into ◽  
Cattle And Sheep

ABSTRACT Analyses of the distribution of virulence factors among different Escherichia coli pathotypes, including Shiga toxin-producing E. coli (STEC), may provide some insight into the mechanisms by which different E. coli strains cause disease and the evolution of distinct E. coli types. The aim of this study was to examine the DNA sequence of the gene for enterohemolysin, a plasmid-encoded toxin that readily causes the hemolysis of washed sheep erythrocytes, and to assess the distribution of enterohemolysin subtypes among E. coli isolates from various human and animal sources. The 2,997-bp ehxA gene was amplified from 227 (63.8%) of 356 stx- and/or eae-positive E. coli strains isolated from cattle and sheep and from 24 (96.0%) of 25 STEC strains isolated from humans with diarrheal disease. By using PCR and restriction fragment length polymorphism (RFLP) analysis of ehxA, six distinct PCR-RFLP types (A to F) were observed, with strains of subtypes A and C constituting 91.6% of all the ehxA-positive strains. Subtype A was associated mainly with ovine strains with stx only (P < 0.001), and subtype C was associated with bovine eae-positive strains (P < 0.001). Eleven ehxA alleles were fully sequenced, and the phylogenetic analysis indicated the presence of three closely related (>95.0%) ehxA sequence groups, one including eae-positive strains (subtypes B, C, E, and F) and the other two including mainly eae-negative STEC strains (subtypes A and D). In addition to being widespread among STEC strains, stx-negative, eae-positive strains (atypical enteropathogenic E. coli strains) isolated from cattle and sheep have similar ehxA subtypes and hemolytic activities.

scholarly journals Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, Brazil

2007 ◽  
Vol 49 (1) ◽  
pp. 41-47 ◽  
Author(s):  
Marcelo Teruyuki Matsumoto ◽  
Ana Marisa Fusco-Almeida ◽  
Lilian Cristiane Baeza ◽  
Márcia de Souza Carvalho Melhem ◽  
Maria José Soares Medes-Giannini
Keyword(s):  
Cryptococcus Neoformans ◽  
Mating Type ◽  
Rflp Analysis ◽  
Sao Paulo ◽  
Clinical Isolates ◽  
São Paulo ◽  
Paulo State ◽  
Pcr Fingerprinting ◽  
Pcr Rflp ◽  
São Paulo State

The basidiomycetous yeast Cryptococcus neoformans is an important fungal pathogen mainly in immunocompromised patients. In this study, 47 clinical isolates of C. neoformans from regions of São Paulo State were studied serologically by using the Crypto Check Iatron RM 304-K kit, their genetic diversity was estimated by PCR-fingerprinting with a microsatellite-specific sequence (GACA)4, RAPD with primer 6 (Amersham Pharmacia Biotech), PCR-restriction fragment length polymorphism (RFLP) analysis of the phospholipase B gene (PLB1) digested with AvaI and mating type analysis by PCR. All 47 strains isolated from HIV positive patients included in this study were serotype A and MATalpha. The majority of the isolates (45/47) were VNI and only two were VNII by PCR-fingerprinting and PCR-RFLP analysis. High degree of homogeneity was observed when (GACA)4 was used, being highly correlated (> 0.9). In contrast, the RAPD analysis was more heterogeneous with higher number of molecular profiles. By PCR-RFLP, no new molecular type was found, enhancing the suggestion that the differences based on conserved gene as PLB1, can be resultant of ongoing divergent evolution within the C. neoformans complex, into the current eight subtypes. Our results furnish new information on the molecular epidemiology of C. neoformans in the southeast region of Brazil.

scholarly journals Novel Hybrid of Typical Enteropathogenic Escherichia coli and Shiga-Toxin-Producing E. coli (tEPEC/STEC) Emerging From Pet Birds

2018 ◽  
Vol 9 ◽  
Author(s):  
Rosely Martins Gioia-Di Chiacchio ◽  
Marcos Paulo Vieira Cunha ◽  
Lilian Rose Marques de Sá ◽  
Yamê Minieiro Davies ◽  
Camila Bueno Pacheco Pereira ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Enteropathogenic Escherichia Coli ◽  
E Coli ◽  
Pet Birds

scholarly journals Pesquisa de enterobacteriáceas e enterovírus em crianças normais e com quadros diarréicos agudos

1968 ◽  
Vol 2 (2) ◽  
pp. 194-206 ◽  
Author(s):  
José Alberto N. Candeias ◽  
Sebastião Timo Iaria ◽  
Dacio de Almeida Christovão ◽  
Ary Walter Schmid ◽  
Augusto de Escragnolle Taunay ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Sao Paulo ◽  
São Paulo ◽  
E Coli

Num grupo de 263 crianças de 0 a 24 meses de idade, atendidas de outubro de 1963 a setembro de 1964 no Centro de Saúde da Lapa, na Capital de São Paulo, foram feitos exames de fezes para a pesquisa de Escherichia coli, do grupo da gastroenterite infantil, Shigella, Salmonella, poliovírus, vírus Coxsackie e vírus ECHO. O agente mais freqüentemente isolado foi o vírus da poliomielite (15,97% de positividade), seguindo-se-lhe a Escherichia coli G.E.I. (10,65%), Shigella (9,51%), vírus Coxsackie (7,22%), Salmonella em 3,04% dos casos e vírus ECHO no mesmo percentual. Em 16,22% das 111 que apresentaram resultado positivo, o exame revelou a presença de dois ou mais agentes. Os agentes acima referidos foram isolados de 42,21% da totalidade das crianças examinadas. Dentre as 167 que apresentavam diarréia, a porcentagem de positividade chegou a 47,90%, e nas 96 que nao a referiam, desceu a 32,29%. Sòmente em relação às shigelas foi possível evidenciar associação entre isolamento de microrganismos e quadro diarréico. Para a E. coli 0111, salmonelas e vírus Coxsackie A, os resultados foram apenas sugestivos desta associação, e para os demais agentes não houve evidência alguma favorável à mesma.

scholarly journals Bactérias gram-negativas em cardeais (Paroaria coronata e Paroaria dominicana) apreendidos do tráfico de animais silvestres

2016 ◽  
Vol 53 (1) ◽  
pp. 107 ◽  
Author(s):  
Marcos Paulo Vieira Cunha ◽  
Marta Brito Guimarães ◽  
Yamê Miniero Davies ◽  
Liliane Milanelo ◽  
Terezinha Knöbl
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Klebsiella Pneumoniae ◽  
Sao Paulo ◽  
São Paulo ◽  
Salmonella Spp ◽  
E Coli

Anualmente o tráfico de animais silvestres retira milhões de aves da natureza. Os cardeais (Paroaria coronata) e cardeais-do-nordeste (Paroaria dominicana) estão incluídos entre as espécies de aves mais traficadas. A microbiota cloacal de passeriformes de vida livre é composta principalmente por bacilos e cocos gram-positivos, já os bacilos gram-negativos predominam em aves de cativeiro. Em situações de estresse e baixa de imunidade as bactérias gram-negativas podem causar infecções oportunistas. O presente trabalho identificou bactérias da microbiota da cloaca de 49 espécimes de P. coronata e P. dominicana apreendidas do tráfico de animais silvestres em São Paulo (SP). Foram isoladas treze espécies de bactérias gram-negativas, incluindo Salmonella spp. e Pseudomonas aeruginosa. A maior frequência de ocorrência foi de Escherichia coli, identificada em 42/49 (85,7%) das amostras fecais. Dentre os isolados de E. coli, 21/42 pertenciam aos grupos filogenéticos B2 e D, relacionados a estirpes patogênicas que causam doença extraintestinal em humanos. Klebsiella pneumoniae foi isolada em 28/49 (57,1%) das amostras. Esses resultados reforçam que as condições estressantes a que esses animais são submetidos em situações de tráfico, incluindo o contato com humanos, podem favorecer a colonização da microbiota cloacal das aves por patógenos, o que representa um risco para a sua reintrodução na natureza considerando-se o possível contato com humanos e outros animais.

scholarly journals Molecular analysis of Escherichia coli O157 strains isolated from cattle and pigs by the use of PCR and pulsed-field gel electrophoresis methods

2012 ◽  
Vol 47 (No. 6) ◽  
pp. 149-158 ◽  
Author(s):  
J. Osek ◽  
P. Gallien
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Genetic Relatedness ◽  
Rflp Analysis ◽  
Toxin Gene ◽  
Escherichia Coli O157 ◽  
Chromosomal Dna ◽  
E Coli ◽  
Shiga Toxin Genes ◽  
Porcine Origin

Fourteen Escherichia coli O157 strains isolated from cattle and pigs in Poland and in Germany were investigated, using PCR, for the genetic markers associated with Shiga toxin-producing E. coli (STEC). Only two strains, both of cattle origin, were positive for the fliC (H7) gene and could be classified as O157 : H7. Nine isolates had stx shiga toxin genes, either stx1 (1 strain), stx2 (4 isolates) or both (4 strains). The stx2-carrying samples were further subtyped by PCR for the stx2c, stx2d, and stx2e toxin variants. It was shown that all but one stx2-positive bacteria possessed the stx2c Shiga toxin gene type and one stx2 STEC isolate had the stx2d virulence factor sub-type. The eaeA (intimin) gene was found in 9 strains (8 isolates from cattle and one strain from pigs); all of them harboured the genetic marker characteristic of the gamma intimin variant. The translocated intimin receptor (tir) gene was detected in 7 isolates tested and among them only one tir-positive strain was recovered from pigs. The ehly E. coli enterohemolysin gene was amplified in all but one strains obtained from cattle and only in one isolate of porcine origin. The genetic relatedness of the analysed E. coli O157 strains was examined by restriction fragment length polymorphism (RFLP) of chromosomal DNA digested with XbaI. Two distinct but related RFLP pattern clusters were observed: one with 9 strains (8 isolates of bovine origin and one strain obtained from pigs) and the other one comprises the remaining 5 E. coli isolates (4 of porcine origin and one strain recovered from cattle). The results suggest that pigs, besides cattle, may be a reservoir of E. coli O157 strains potentially pathogenic to humans. Moreover, epidemiologically unrelated isolates of the O157 serogroup, recovered from different animal species, showed a clonal relationship as demonstrated by the RFLP analysis.

scholarly journals Origin of Contamination and Genetic Diversity of Escherichia coli in Beef Cattle

2003 ◽  
Vol 69 (5) ◽  
pp. 2794-2799 ◽  
Author(s):  
Mueen Aslam ◽  
Frances Nattress ◽  
Gordon Greer ◽  
Chris Yost ◽  
Colin Gill ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Genetic Diversity ◽  
Beef Cattle ◽  
Ground Beef ◽  
Rflp Analysis ◽  
E Coli ◽  
Cattle Feces ◽  
Genetic Subtypes ◽  
Random Amplification ◽  
Pcr Rflp

ABSTRACT The possible origin of beef contamination and genetic diversity of Escherichia coli populations in beef cattle, on carcasses and ground beef, was examined by using random amplification of polymorphic DNA (RAPD) and PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of the fliC gene. E. coli was recovered from the feces of 10 beef cattle during pasture grazing and feedlot finishing and from hides, carcasses, and ground beef after slaughter. The 1,403 E. coli isolates (855 fecal, 320 hide, 153 carcass, and 75 ground beef) were grouped into 121 genetic subtypes by using the RAPD method. Some of the genetic subtypes in cattle feces were also recovered from hides, prechilled carcasses, chilled carcasses, and ground beef. E. coli genetic subtypes were shared among cattle at all sample times, but a number of transient types were unique to individual animals. The genetic diversity of the E. coli population changed over time within individual animals grazing on pasture and in the feedlot. Isolates from one animal (59 fecal, 30 hide, 19 carcass, and 12 ground beef) were characterized by the PCR-RFLP analysis of the fliC gene and were grouped into eight genotypes. There was good agreement between the results obtained with the RAPD and PCR-RFLP techniques. In conclusion, the E. coli contaminating meat can originate from cattle feces, and the E. coli population in beef cattle was highly diverse. Also, genetic subtypes can be shared among animals or can be unique to an animal, and they are constantly changing.

scholarly journals Re-emergence of O103 : H2 Shiga toxin-producing Escherichia coli infections in São Paulo, Brazil

2005 ◽  
Vol 54 (8) ◽  
pp. 805-806 ◽  
Author(s):  
Beatriz EC Guth ◽  
Tânia MI Vaz ◽  
Tânia AT Gomes ◽  
Silvia H Chinarelli ◽  
Marilu MM Rocha ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Sao Paulo ◽  
São Paulo
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