Res. z~crobioL 1990, 141, 703-706
© INSTITUTPASTEUR/ELSEVIER Paris 1990
P R E L I M I N A R Y NOTE
IDENTIFICATION OF A NEW ENTEROINVASIVE
E S C H E R I C H I A COLI STRAIN
D.M.M. Queiroz (~), E.N. Mendes (1), M.R.F. Toledo (7) and L.R. Trabulsi (2) (l) Laboratory of Research in Bacteriology, Faculdae2e de Medicina, Universidade Federal de Minas Gerais, Av. Aifredo Balen~, 190/464, CEP 30130, Belo Horizonte, MG (Brazil), and (2) Department of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina, Rua Botucatu, 862, CEP 04023, Sa6 t~auio, SP (Brazil)
SUMMARY We isolated an enteroinvasive Escherichia coli strain which did not react with antisera raised against all known E. coli serogroups, Shigella groups A, B, C and D er provisional Shigella serotypes. We propose the provisional designation of Escherichia coli MG for this strain, until the identification of its 0 antigen is, established. KEY-WORDS: Escherichia coli MG, Enteritis; New strain.
Enteroinvasive Escherichia coli strains are enteropathogenic and cause a Shigella-like disease in children and adults which is characterized by fever, chills, cramping abdominal pain, tenesmus and small-volume stools containing blood, Ira's and mucus (Carpenter, 1983 ; W H O Scientific Working Group, 1980). Most of them have O antigens identical or strongly related to O antigens of some serotypes of Shigella dysenteriae and Shigella boydii (Ewing, 1986). Furthermore, both Shigella and enteroinvasive E. coil cause keratocon-
Submitted January 12, 1990, accepted June 20, 1990.
D.M.M. QUEIROZ E T AL.
704 TABLr, I. - -
Biochemical reactions of the i n v a s i v e E. coli i s o l a t e . Test
lndole (Kovacs) Methyl red Voges-Proskauer Christensen's citrate Simmons' citrate Christensen's urea Phenylalanine deaminase HTS ( T S I ) N~trate, reduction to nitrite Lysine decarboxylase Arginine dihydrolase Ornithine decarboxylase Motility (36°C) Gelatin liquefaction (22°C) Growth in KCN Malonate utilization D-glucose (gas)
Reaction + + + (~) +
Acid production from: D-glucose Lactose D-mannose Sucrose D-mannitoi Dulcitol Salicin Adonitol i-Inositol D-sorbitol L-arabinose Raffinose L-rhamnose Maltose D-xylose Trehalose Cellobiose Erythritol Melibiose D-arabitol Glycerol Mucate Jordan's tartrate • -CHa-glucoside Escuhn Acetate L i p a s e ( c o r n oil) DNase (25°C) Oxidase (Kovacs) Galactosidase (ONPG) - g l u c o s e f e r m e n t e d in O F m e d i u m Pectate liquefaction
~
+ (+ ) + + + + + + + + + + (+ ) + + + -
+ = positive reaction within ! or 2 days of incubation ; - = negative reaction; ( + ) = positive reaction after 3 or more days of incubation. TSI = triple sugar iron; ONPG = o-nitrophenyl-[~-D-galactopyranoside.
A N E W E N T E R O I N V A S I V E E. COLI
705
junctivitis in the guinea pig (Ser6ny test) (Ser6ny, 1955 ; Trabulsi et al., 1967; WHO Scientific Working Group, 1980). All of the described isolates of enteroinvasive E. coil strains do not decarboxylate lysine and are non-motile, except some strains of the O124 and O144 serogroups that are motile (Silva et al., 1980; Toledo and Trabulsi, 1983; Gomes et al., 1987). The O serogtoups to which enteroinvasive E. coil strains belong were gradually expanded and now include O28ac, 029, O112ac, O124, O136, O143, O144, Ot52, O164 and O167 (Gomes et al., 1987; Toledo and Trabulsi, 1983). According to Gross et al. (1983), recognition of new enteroinvasive serogroups facilitates the determination of the epidemiology of diarrhoeal diseases caused by enteroinvasive E. coil which are not well defined. While investigating the aetiology of adult diarrhoea in Belo Horizonte (Brazil), we isolated an E. coil strain (4308-88) from the faeces of a 22-year old female which was strongly positive in the Ser6ny test. It was the only pathogen found. Biochemical tests carried out for identifying this strain, using methodology recommended by Ewing 0986), demonstrated that it was a delayed lactose-positive non-motile E. coil which did not decarboxylate lysine (table I). Furthermore, the strain did not agglutinate with O antisera prepared against en.:erc,invasive E. coli serogroups O28ac, 029, O112ac, O124, O136, O143, O144, O152, O164 and O167 as well as against S. dysenteriae 1-12, S. flexneri 1-6, S. boydii 1-18 and S. sonnei phases I and II. In disk agar diffusion test (Bauer et al., 1966), the isolate was susceptible to sulfamethoxazole÷ trimethoprim, chloramphenicol, cephalothin, ampicillin, colistin, gentamicin, neomycin, kanamycin and amikacin, and was resistant to erythromycin. This strain was sent to the Enteric Section of the Centers for Disease Control (CDC), Atlanta, GA, USA (CDC no. 3173-88). Biochemical reactions, serological agglutination tests as well as Ser6ny test and an ELISA test for invasiveness performed at CDC confirmed our results. In addition, the strain did not react with antisera raised against all previously described E. coil O group (O1 to O171) and Shigella provisional serotypes 1621-54 and E16553. The results obtained point out that this isolate belongs to a new invasive E. coil type which has an undescribed O antigen. We propose the designation of E. coil MG for this serotype until the status of its O antigen is established and it is included in the E. coil 0 antigen scheme. We are producing an antiserum against thi~ strain by employing the method recommended by Ewing (1986) which will enable the identification of other isolates and thus assess the prevalence of this new strain in different geographical areas.
706
D.M.M. QUEIROZ ET AL.
RI~.SUMI~ IDENTIFICATION D'UN NOUVEAU TYPE DE ESCHERICHIA COL! ENTEROINVASIF
Nous avons isole une souche enteroinvasive de Escherichia coil qui ne reagit avec aucun des immunserums d i r i # contre les s6rogroupes de E. coil, contre les groupes A, B, C et D de Shigella et contre les serotypes provisoires de Shigella. Nous proposons la designation temporaire de cette souche comme Escherichia coli MG, jusqu'/~ ce que le profil de ses antig~nes O suit etabli. MOTS.CLI~S: Escherichia coli MG, Enterite; Nouvelle souche.
ACKNOWLEDGEMENTS We thank Dr. Paul A. Blake from Enteric Diseases Branch, CDC, Atlanta, GA (USA) for confirmation of the biochemical, serological and invasive properties of the strain. This s'~udy was supported by grants from Conselho Nacional de Desenvolvimento Cientifico e Tecnol6gico (CNPq) and Financiadora de Estudos e Projetos (FINEP) (Brazil).
REFERENCES BAUER, A.W., KIRBY, W.M.M., SHERRIS,T.C. & TURCK, M. (1966), Antibiotic susceptibility testing by a standardized single disk method. Amer. J. Olin. Path., 45, 493-496. CARPENTER, C.C.J. (1983), Diarrheal disease caused by Escherichia coil, in "Infectious Diseases" (P.D. Hoeprich), 3 (pp. 643-648). Harper & Row, Philadelphia. EWIN~3, W.H. (1986), "Edwards and Ewing's Identification of Enterobacteriaceae", 4, Elsevie~ Science Publ., Amsterdam. GOMES, T.A.T., TOLEDO,M.R.F., TRABULSI,L.R., WOOD, P.K. & MoRRiS, J.G. Jr. (1987), DNA l~robes for identification of enteroinvasive Escherichia coli. J. clin. Microbiol., 25, 2025-2027. GROSS, R.J., THOMAS, L.V., CHEASTI, T., DAY, N.P., ROWE, B., TOLEDO, M.R.F. & TRABOLSl, L.R. (1983), Enterotoxigenic and enteroinvasive Escherichia coil strain belonging to a new O group, O167. J. clin. MicrobioL, 17, 521-523. SER~NY, B. (1955), Experimental Shigeila keratoconjunctivitis. A preliminary report. Acta. microbiol. Acad. Sci. Hung., 2, 293-296. SILVA, R.M., TOLEDO, M.R.F. & TRABULSt, L.R. (1980), Biochemical and cultural characteristics of invasive Escherichia coll. J. clin. MicrobioL, 11, 441-444. TOLEDO, M.R.F. & TRABULSl, L.R. (1983), Correlation between biochemical and serological characteristics of Escherichia coil and results of the Sereny test. J. clia. Microbiol., 1"/, 419-421. TRABULSl,L.R., TOLEOO,M.R.F. & ZULIANI,M.E. (1967), Novas bacterias patogSnicas parR o intestino do homem. Rev. Inst. Med. trop. (S. Paulo), 9, 31-39. WHO SCIENTIFIC WORKING GROUP (1980), Escherichia coli diarrhoea. Bull. Org. round. SanM, 58, 23-36.
© INSTITUTPASTEUR/ELSEVIER Paris 1990
P R E L I M I N A R Y NOTE
IDENTIFICATION OF A NEW ENTEROINVASIVE
E S C H E R I C H I A COLI STRAIN
D.M.M. Queiroz (~), E.N. Mendes (1), M.R.F. Toledo (7) and L.R. Trabulsi (2) (l) Laboratory of Research in Bacteriology, Faculdae2e de Medicina, Universidade Federal de Minas Gerais, Av. Aifredo Balen~, 190/464, CEP 30130, Belo Horizonte, MG (Brazil), and (2) Department of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina, Rua Botucatu, 862, CEP 04023, Sa6 t~auio, SP (Brazil)
SUMMARY We isolated an enteroinvasive Escherichia coli strain which did not react with antisera raised against all known E. coli serogroups, Shigella groups A, B, C and D er provisional Shigella serotypes. We propose the provisional designation of Escherichia coli MG for this strain, until the identification of its 0 antigen is, established. KEY-WORDS: Escherichia coli MG, Enteritis; New strain.
Enteroinvasive Escherichia coli strains are enteropathogenic and cause a Shigella-like disease in children and adults which is characterized by fever, chills, cramping abdominal pain, tenesmus and small-volume stools containing blood, Ira's and mucus (Carpenter, 1983 ; W H O Scientific Working Group, 1980). Most of them have O antigens identical or strongly related to O antigens of some serotypes of Shigella dysenteriae and Shigella boydii (Ewing, 1986). Furthermore, both Shigella and enteroinvasive E. coil cause keratocon-
Submitted January 12, 1990, accepted June 20, 1990.
D.M.M. QUEIROZ E T AL.
704 TABLr, I. - -
Biochemical reactions of the i n v a s i v e E. coli i s o l a t e . Test
lndole (Kovacs) Methyl red Voges-Proskauer Christensen's citrate Simmons' citrate Christensen's urea Phenylalanine deaminase HTS ( T S I ) N~trate, reduction to nitrite Lysine decarboxylase Arginine dihydrolase Ornithine decarboxylase Motility (36°C) Gelatin liquefaction (22°C) Growth in KCN Malonate utilization D-glucose (gas)
Reaction + + + (~) +
Acid production from: D-glucose Lactose D-mannose Sucrose D-mannitoi Dulcitol Salicin Adonitol i-Inositol D-sorbitol L-arabinose Raffinose L-rhamnose Maltose D-xylose Trehalose Cellobiose Erythritol Melibiose D-arabitol Glycerol Mucate Jordan's tartrate • -CHa-glucoside Escuhn Acetate L i p a s e ( c o r n oil) DNase (25°C) Oxidase (Kovacs) Galactosidase (ONPG) - g l u c o s e f e r m e n t e d in O F m e d i u m Pectate liquefaction
~
+ (+ ) + + + + + + + + + + (+ ) + + + -
+ = positive reaction within ! or 2 days of incubation ; - = negative reaction; ( + ) = positive reaction after 3 or more days of incubation. TSI = triple sugar iron; ONPG = o-nitrophenyl-[~-D-galactopyranoside.
A N E W E N T E R O I N V A S I V E E. COLI
705
junctivitis in the guinea pig (Ser6ny test) (Ser6ny, 1955 ; Trabulsi et al., 1967; WHO Scientific Working Group, 1980). All of the described isolates of enteroinvasive E. coil strains do not decarboxylate lysine and are non-motile, except some strains of the O124 and O144 serogroups that are motile (Silva et al., 1980; Toledo and Trabulsi, 1983; Gomes et al., 1987). The O serogtoups to which enteroinvasive E. coil strains belong were gradually expanded and now include O28ac, 029, O112ac, O124, O136, O143, O144, Ot52, O164 and O167 (Gomes et al., 1987; Toledo and Trabulsi, 1983). According to Gross et al. (1983), recognition of new enteroinvasive serogroups facilitates the determination of the epidemiology of diarrhoeal diseases caused by enteroinvasive E. coil which are not well defined. While investigating the aetiology of adult diarrhoea in Belo Horizonte (Brazil), we isolated an E. coil strain (4308-88) from the faeces of a 22-year old female which was strongly positive in the Ser6ny test. It was the only pathogen found. Biochemical tests carried out for identifying this strain, using methodology recommended by Ewing 0986), demonstrated that it was a delayed lactose-positive non-motile E. coil which did not decarboxylate lysine (table I). Furthermore, the strain did not agglutinate with O antisera prepared against en.:erc,invasive E. coli serogroups O28ac, 029, O112ac, O124, O136, O143, O144, O152, O164 and O167 as well as against S. dysenteriae 1-12, S. flexneri 1-6, S. boydii 1-18 and S. sonnei phases I and II. In disk agar diffusion test (Bauer et al., 1966), the isolate was susceptible to sulfamethoxazole÷ trimethoprim, chloramphenicol, cephalothin, ampicillin, colistin, gentamicin, neomycin, kanamycin and amikacin, and was resistant to erythromycin. This strain was sent to the Enteric Section of the Centers for Disease Control (CDC), Atlanta, GA, USA (CDC no. 3173-88). Biochemical reactions, serological agglutination tests as well as Ser6ny test and an ELISA test for invasiveness performed at CDC confirmed our results. In addition, the strain did not react with antisera raised against all previously described E. coil O group (O1 to O171) and Shigella provisional serotypes 1621-54 and E16553. The results obtained point out that this isolate belongs to a new invasive E. coil type which has an undescribed O antigen. We propose the designation of E. coil MG for this serotype until the status of its O antigen is established and it is included in the E. coil 0 antigen scheme. We are producing an antiserum against thi~ strain by employing the method recommended by Ewing (1986) which will enable the identification of other isolates and thus assess the prevalence of this new strain in different geographical areas.
706
D.M.M. QUEIROZ ET AL.
RI~.SUMI~ IDENTIFICATION D'UN NOUVEAU TYPE DE ESCHERICHIA COL! ENTEROINVASIF
Nous avons isole une souche enteroinvasive de Escherichia coil qui ne reagit avec aucun des immunserums d i r i # contre les s6rogroupes de E. coil, contre les groupes A, B, C et D de Shigella et contre les serotypes provisoires de Shigella. Nous proposons la designation temporaire de cette souche comme Escherichia coli MG, jusqu'/~ ce que le profil de ses antig~nes O suit etabli. MOTS.CLI~S: Escherichia coli MG, Enterite; Nouvelle souche.
ACKNOWLEDGEMENTS We thank Dr. Paul A. Blake from Enteric Diseases Branch, CDC, Atlanta, GA (USA) for confirmation of the biochemical, serological and invasive properties of the strain. This s'~udy was supported by grants from Conselho Nacional de Desenvolvimento Cientifico e Tecnol6gico (CNPq) and Financiadora de Estudos e Projetos (FINEP) (Brazil).
REFERENCES BAUER, A.W., KIRBY, W.M.M., SHERRIS,T.C. & TURCK, M. (1966), Antibiotic susceptibility testing by a standardized single disk method. Amer. J. Olin. Path., 45, 493-496. CARPENTER, C.C.J. (1983), Diarrheal disease caused by Escherichia coil, in "Infectious Diseases" (P.D. Hoeprich), 3 (pp. 643-648). Harper & Row, Philadelphia. EWIN~3, W.H. (1986), "Edwards and Ewing's Identification of Enterobacteriaceae", 4, Elsevie~ Science Publ., Amsterdam. GOMES, T.A.T., TOLEDO,M.R.F., TRABULSI,L.R., WOOD, P.K. & MoRRiS, J.G. Jr. (1987), DNA l~robes for identification of enteroinvasive Escherichia coli. J. clin. Microbiol., 25, 2025-2027. GROSS, R.J., THOMAS, L.V., CHEASTI, T., DAY, N.P., ROWE, B., TOLEDO, M.R.F. & TRABOLSl, L.R. (1983), Enterotoxigenic and enteroinvasive Escherichia coil strain belonging to a new O group, O167. J. clin. MicrobioL, 17, 521-523. SER~NY, B. (1955), Experimental Shigeila keratoconjunctivitis. A preliminary report. Acta. microbiol. Acad. Sci. Hung., 2, 293-296. SILVA, R.M., TOLEDO, M.R.F. & TRABULSt, L.R. (1980), Biochemical and cultural characteristics of invasive Escherichia coll. J. clin. MicrobioL, 11, 441-444. TOLEDO, M.R.F. & TRABULSl, L.R. (1983), Correlation between biochemical and serological characteristics of Escherichia coil and results of the Sereny test. J. clia. Microbiol., 1"/, 419-421. TRABULSl,L.R., TOLEOO,M.R.F. & ZULIANI,M.E. (1967), Novas bacterias patogSnicas parR o intestino do homem. Rev. Inst. Med. trop. (S. Paulo), 9, 31-39. WHO SCIENTIFIC WORKING GROUP (1980), Escherichia coli diarrhoea. Bull. Org. round. SanM, 58, 23-36.
