CC BY-NC-ND 4.0 · J Lab Physicians 2019; 11(04): 346-351
DOI: 10.4103/JLP.JLP_58_19
Original Article

Evolution of diarrheagenic Escherichia coli pathotypes in India

Pankaj Singh
Department of Medical Microbiology, All India Institute of Medical Sciences, Bhopal, Madhya Pradesh, India
,
Sharda C. Metgud
Department of Microbiology, Jawaharlal Nehru Medical College, Belgaum, Karnataka, India
,
Subarna Roy
RMRC, ICMR, Belgaum, Karnataka, India
,
Shashank Purwar
Department of Medical Microbiology, All India Institute of Medical Sciences, Bhopal, Madhya Pradesh, India
› Author Affiliations
Financial support and sponsorship Nil.

Abstract

CONTEXT: Diarrheagenic Escherichia coli (DEC) is the leading cause of infectious diarrhea in developing countries. On the basis of virulence and phenotypic characteristics, the DEC is categorized into multiple pathotypes. Each pathotype has different pathogenesis and geographical distribution. Thus, the proper management of disease relies on rapid and accurate identification of DEC pathotypes.

AIMS: The aim of the study was to determine the prevalence of DEC pathotypes in India.

MATERIALS AND METHODS: A cross-sectional study was carried out between January 2008 and December 2012 at Jawaharlal Nehru Medical College and KLES Dr. Prabhakar Kore Hospital and Medical Research Center, Belgaum (Karnataka), India. A total of 300 stool samples were collected from diarrhea patients with age >3 months. The DEC was identified by both conventional and molecular methods.

RESULTS: Of 300 samples, E. coli were detected in 198 (66%) and 170 (56.6%) samples by culture and polymerase chain reaction, respectively. Among DEC (n = 198) isolates, eae gene (59.5%) was the most prevalent followed by stx (27.7%), east (27.2%), elt (12.6%), est (10.6%), ipaH (5.5%), and eagg (1.5%) genes. On the basis of virulence genes, enteropathogenic E. coli (33.8%) was the most common pathotype followed by Shiga toxin-producing E. coli (STEC, 23.2%), enterotoxigenic E. coli (ETEC, 13.6%), enteroinvasive E. coli (5.5%), enteroaggregative heat-stable enterotoxin 1-harboring E. coli (EAST1EC, 4.5%), STEC/ETEC (3.5%), STEC/enteroaggregative E. coli (STEC/EAEC, 1.0%), and EAEC (0.05%).

CONCLUSIONS: The hybrid DEC is potentially more virulent than basic pathotypes. The pathotyping should be included in clinical settings for the proper management of DEC-associated diarrhea.



Publication History

Received: 06 April 2019

Accepted: 10 October 2019

Article published online:
07 April 2020

© 2019.

Thieme Medical and Scientific Publishers Private Ltd.
A-12, Second Floor, Sector -2, NOIDA -201301, India

 
  • References

  • 1 Kotloff KL, Blackwelder WC, Nasrin D, Nataro JP, Farag TH, van Eijk A, et al. The global enteric multicenter study (GEMS) of diarrheal disease in infants and young children in developing countries: Epidemiologic and clinical methods of the case/control study. Clin Infect Dis 2012;55 Suppl 4:S232-45.
  • 2 Mladenova Z, Steyer A, Steyer AF, Ganesh B, Petrov P, Tchervenjakova T, et al. Aetiology of acute paediatric gastroenteritis in Bulgaria during summer months: Prevalence of viral infections. J Med Microbiol 2015;64:272-82.
  • 3 Shrivastava AK, Kumar S, Mohakud NK, Suar M, Sahu PS. Multiple etiologies of infectious diarrhea and concurrent infections in a pediatric outpatient-based screening study in Odisha, India. Gut Pathog 2017;9:16.
  • 4 Saeed A, Abd H, Sandstrom G. Microbial aetiology of acute diarrhoea in children under five years of age in Khartoum, Sudan. J Med Microbiol 2015;64:432-7.
  • 5 Nataro JP, Kaper JB. Diarrheagenic Escherichia coli. Clin Microbiol Rev 1998;11:142-201.
  • 6 Gomes TA, Elias WP, Scaletsky IC, Guth BE, Rodrigues JF, Piazza RM, et al. Diarrheagenic Escherichia coli. Braz J Microbiol 2016;47 Suppl 1:3-0.
  • 7 Dalla-Costa LM, Irino K, Rodrigues J, Rivera IN, Trabulsi LR. Characterisation of diarrhoeagenic Escherichia coli clones by ribotyping and ERIC-PCR. J Med Microbiol 1998;47:227-34.
  • 8 Hegde A, Ballal M, Shenoy S. Detection of diarrheagenic Escherichia coli by multiplex PCR. Indian J Med Microbiol 2012;30:279-84.
  • 9 Alizade H, Fallah F, Ghanbarpour R, Aflatoonian MR, Taherpour A, Sharifi H, et al. Genotyping of ESBL producing uropathogenic and diarrheagenic Escherichia coli in southeast of Iran. Infect Disord Drug Targets 2015;15:118-24.
  • 10 Hosseini Nave H, Mansouri S, Taati Moghadam M, Moradi M. Virulence gene profile and multilocus variable-number tandem-repeat analysis (MLVA) of enteroinvasive Escherichia coli (EIEC) isolates from patients with diarrhea in Kerman, Iran. Jundishapur J Microbiol 2016;9:e33529.
  • 11 Yu F, Chen X, Zheng S, Han D, Wang Y, Wang R, et al. Prevalence and genetic diversity of human diarrheagenic Escherichia coli isolates by multilocus sequence typing. Int J Infect Dis 2018;67:7-13.
  • 12 Toma C, Lu Y, Higa N, Nakasone N, Chinen I, Baschkier A, et al. Multiplex PCR assay for identification of human diarrheagenic Escherichia coli. J Clin Microbiol 2003;41:2669-71.
  • 13 Kimata K, Shima T, Shimizu M, Tanaka D, Isobe J, Gyobu Y, et al. Rapid categorization of pathogenic Escherichia coli by multiplex PCR. Microbiol Immunol 2005;49:485-92.
  • 14 Fujioka M, Otomo Y, Ahsan CR. A novel single-step multiplex polymerase chain reaction assay for the detection of diarrheagenic Escherichia coli. J Microbiol Meth 2013;92:289-92.
  • 15 Mora A, Herrrera A, López C, Dahbi G, Mamani R, Pita JM, et al. Characteristics of the shiga-toxin-producing enteroaggregative Escherichia coli O104:H4 German outbreak strain and of STEC strains isolated in Spain. Int Microbiol 2011;14:121-41.
  • 16 Nyholm O, Halkilahti J, Wiklund G, Okeke U, Paulin L, Auvinen P, et al. Comparative genomics and characterization of hybrid shigatoxigenic and enterotoxigenic Escherichia coli (STEC/ETEC) strains. PLoS One 2015;10:e0135936.
  • 17 Savarino SJ, Fasano A, Robertson DC, Levine MM. Enteroaggregative Escherichia coli elaborate a heat-stable enterotoxin demonstrable in an in vitro rabbit intestinal model. J Clin Invest 1991;87:1450-55.
  • 18 Yamamoto T, Wakisaka N, Sato F, Kato A. Comparison of the nucleotide sequence of enteroaggregative Escherichia coli heat-stable enterotoxin 1 genes among diarrhea-associated Escherichia coli. FEMS Microbiol Lett 1997;147:89-95.
  • 19 Ruan X, Crupper SS, Schultz BD, Robertson DC, Zhang W. Escherichia coli expressing EAST1 toxin did not cause an increase of cAMP or cGMP levels in cells, and no diarrhea in 5-day old gnotobiotic pigs. PloS one 2012;7:e43203.
  • 20 Zajacova ZS, Konstantinova L, Alexa P. Detection of virulence factors of Escherichia coli focused on prevalence of EAST1 toxin in stool of diarrheic and non-diarrheic piglets and presence of adhesion involving virulence factors in astA positive strains. Vet Microbiol 2012;154:369-75.
  • 21 Silva LE, Souza TB, Silva NP, Scaletsky IC. Detection and genetic analysis of the enteroaggregative Escherichia coli heat-stable enterotoxin (EAST1) gene in clinical isolates of enteropathogenic Escherichia coli (EPEC) strains. BMC Microbiol 2014;14:135.
  • 22 Minas K, McEwan NR, Newbold CJ, Scott KP. Optimization of a high-throughput CTAB-based protocol for the extraction of qPCR-grade DNA from rumen fluid, plant and bacterial pure cultures. FEMS Microbiol Lett 2011;325:162-9.
  • 23 Schmidt H, Knop C, Franke S, Aleksic S, Heesemann J, Karch H. Development of PCR for screening of enteroaggregative Escherichia coli. J Clin Microbiol 1995;33:701-5.
  • 24 Chakraborty S, Deokule JS, Garg P, Bhattacharya SK, Nandy RK, Nair GB, et al. Concomitant infection of enterotoxigenic Escherichia coli in an outbreak of cholera caused by Vibrio cholerae O1 and O139 in Ahmedabad, India. J Clin Microbiol 2001;39:3241-6.
  • 25 Paciorek J. Virulence properties of Escherichia coli faecal strains isolated in Poland from healthy children and strains belonging to serogroups O18, O26, O44, O86, O126 and O127 isolated from children with diarrhoea. J Med Microbiol 2002;51:548-56.
  • 26 Kahali S, Sarkar B, Rajendran K, Khanam J, Yamasaki S, Nandy RK, et al. Virulence characteristics and molecular epidemiology of enteroaggregative Escherichia coli isolates from hospitalized diarrheal patients in Kolkata, India. J Clin Microbiol 2004;42;4111-20.
  • 27 Bischoff C, Luthy J, Altwegg M, Baggi F. Rapid detection of diarrheagenic E. coli by real-time PCR. J Microbiol Methods 2005;61:335-41.
  • 28 Blank TE, Zhong H, Bell AL, Whittam TS, Donnenberg MS. Molecular variation among type IV pilin (bfpA) genes from diverse enteropathogenic Escherichia coli strains. Infect Immun 2000;68:7028-38.
  • 29 Thakur N, Jain S, Changotra H, Shrivastava R, Kumar Y, Grover N, et al. Molecular characterization of diarrheagenic Escherichia coli pathotypes: Association of virulent genes, serogroups, and antibiotic resistance among moderate-to-severe diarrhea patients. J Clin Lab Anal 2018;32:e22388.
  • 30 Chandra BK, Singh G, Taneja N, Pahil S, Singhi S, Sharma M. Diarrhoeagenic Escherichia coli as a predominant cause of paediatric nosocomial diarrhoea in India. J Med Microbiol 2012;61:830-6.
  • 31 Sethabutr O, Venkatesan M, Murphy GS, Eampokalap B, Hoge CW, Echeverria P, et al. Detection of shigellae and enteroinvasive Escherichia coli by amplification of the invasion plasmid antigen H DNA sequence in patients with dysentery. J Infect Dis 1993;167:458-61.
  • 32 Navarro-Garcia F, Elias WP. Autotransporters and virulence of enteroaggregative E. coli. Gut Microbes 2011;2:13-24.
  • 33 Pabst WL, Altwegg M, Kind C, Mirjanic S, Hardegger D, Nadal D. Prevalence of enteroaggregative Escherichia coli among children with and without diarrhea in Switzerland. J Clin Microbiol 2003;41:2289-93.
  • 34 Sidhu JP, Ahmed W, Hodgers L, Toze S. Occurrence of virulence genes associated with Diarrheagenic pathotypes in Escherichia coli isolates from surface water. Applied and environmental microbiology 2013;79:328-35.
  • 35 Nishikawa Y, Ogasawara J, Helander A, Haruki K. An outbreak of gastroenteritis in Japan due to Escherichia coli O166. Emerg Infect Dis 1999;5:300.
  • 36 Nishikawa Y, Zhou Z, Hase A, Ogasawara J, Kitase T, Abe N, et al. Diarrheagenic Escherichia coli isolated from stools of sporadic cases of diarrheal illness in Osaka city, Japan between 1997 and 2000: Prevalence of enteroaggregative E. coli heat-stable enterotoxin 1 gene-possessing E. coli. Jpn J Infect Dis 2002;55:183-90.
  • 37 Dutta S, Pazhani GP, Nataro JP, Ramamurthy T. Heterogenic virulence in a diarrheagenic Escherichia coli: Evidence for an EPEC expressing heat-labile toxin of ETEC. Int J Med Microbiol 2015;305:47-54.
  • 38 Rasko DA, Webster DR, Sahl JW, Bashir A, Boisen N, Scheutz F, et al. Origins of the E. coli strain causing an outbreak of hemolytic-uremic syndrome in Germany. N Engl J Med 2011;365:709-17.
  • 39 Gunzburg ST, Chang BJ, Elliott SJ, Burke V, Gracey M. Diffuse and enteroaggregative patterns of adherence of enteric Escherichia coli isolated from aboriginal children from the Kimberley region of Western Australia. J Infect Dis 1993;167:755-8.