Comparative analysis of virulence determinants, phylogroups, and antibiotic susceptibility patterns of typical versus atypical Enteroaggregative E. coli in India.

Vinay Modgil,Manish Kakkar,Kawaljeet Singh Sandha,Pankaj Chaudhary,Chandradeo Narayan,Seema Tanwar,Jaspreet Mahindroo,Kamlesh Thakur,Neelam Taneja,Manmohit Kalia,Varun Shahi,Balvinder Mohan,Meenakshi Koundal,Digvijay Singh,Bhavneet Bharti,Neha Gautam,Pratima Gupta,Yousuf ,Rajeev Jain

doi:10.1371/journal.pntd.0008769

Abstract

Enteroaggregative Escherichia coli (EAEC) is an evolving enteric pathogen that causes acute and chronic diarrhea in developed and industrialized nations in children. EAEC epidemiology and the importance of atypical EAEC (aEAEC) isolation in childhood diarrhea are not well documented in the Indian setting. A comparative analysis was undertaken to evaluate virulence, phylogeny, and antibiotic sensitivity among typical tEAEC versus aEAEC. A total of 171 EAEC isolates were extracted from a broad surveillance sample of diarrheal (N = 1210) and healthy children (N = 550) across North India. Polymerase chain reaction (PCR) for the aggR gene (master regulator gene) was conducted to differentiate tEAEC and aEAEC. For 21 virulence genes, we used multiplex PCR to classify possible virulence factors among these strains. Phylogenetic classes were identified by a multiplex PCR for chuA, yjaA, and a cryptic DNA fragment, TspE4C2. Antibiotic susceptibility was conducted by the disc diffusion method as per CLSI guidelines. EAEC was associated with moderate to severe diarrhea in children. The prevalence of EAEC infection (11.4%) was higher than any other DEC group (p = 0.002). tEAEC occurrence in the diarrheal group was higher than in the control group (p = 0.0001). tEAEC strain harbored more virulence genes than aEAEC. astA, aap, and aggR genes were most frequently found in the EAEC from the diarrheal population. Within tEAEC, this gene combination was present in more than 50% of strains. Also, 75.8% of EAEC strains were multidrug-resistant (MDR). Phylogroup D (43.9%) and B1 (39.4%) were most prevalent in the diarrheal and control group, respectively. Genetic analysis revealed EAEC variability; the comparison of tEAEC and aEAEC allowed us to better understand the EAEC virulence repertoire. Further microbiological and epidemiological research is required to examine the pathogenicity of not only typical but also atypical EAEC.

Highlights

Diarrhea has been reported as the second most common cause of mortality all over the world in children less than 5 years of age, causing approximately 1.5 million pediatric deaths per year (WHO/UNICEF, 2009)
Genes that could contribute to the pathogenicity of Enteroaggregative E. coli (EAEC) encode adhesions, toxins, and other factors
AstA, aap, and aggR genes were most frequently found in the EAEC from the diarrheal population

Summary

Introduction

Diarrhea has been reported as the second most common cause of mortality all over the world in children less than 5 years of age, causing approximately 1.5 million pediatric deaths per year (WHO/UNICEF, 2009). Diarrheogenic E. coli (DEC) are the strains of E. coli that, by certain virulence factors, lead to diarrhea in humans. DEC has been categorized into six major pathotypes, including Enteropathogenic E. coli (EPEC), Enterotoxigenic E. coli (ETEC), Enteroaggregative E. coli (EAEC), Enteroinvasive E. coli (EIEC), Enterohemorrhagic E.coli (EHEC), and Diffusely Adherent E. coli (DAEC). EAEC is amongst the major bacterial causes of diarrhea in children worldwide. EAEC is thought to be the most recently identified pathotype among DEC [1]. This pathotype is significantly associated with acute and persistent diarrhea, malnutrition in children and HIV infection, and traveler’s diarrhea [2,3,4,5]. The distribution of EAEC varies from one geographic location to another [6]

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