Genotypic and Phenotypic Characterization of Invasive Neonatal Escherichia coli Clinical Isolates

Salika Mehreen Shakir; Jessica Marie Goldbeck; Denise Robison; Annette Marie Eckerd; Susana Chavez-Bueno

doi:10.1055/s-0034-1370341

American Journal of Perinatology, Table of Contents

Am J Perinatol 2014; 31(11): 975-982
DOI: 10.1055/s-0034-1370341

Original Article

Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Genotypic and Phenotypic Characterization of Invasive Neonatal Escherichia coli Clinical Isolates

Salika Mehreen Shakir

¹Department of Pediatrics, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma

,

Jessica Marie Goldbeck

¹Department of Pediatrics, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma

,

Denise Robison

²Clinical Microbiology Laboratory, OU Medical Center, Oklahoma City, Oklahoma

,

Annette Marie Eckerd

¹Department of Pediatrics, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma

,

Susana Chavez-Bueno

¹Department of Pediatrics, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma

› Author Affiliations

Abstract

Objectives The objective of this study was to describe the clinical characteristics of neonates with Escherichia coli bacteremia and the antibiotic resistance pattern of the bacterial isolates. We assessed the isolates' genetic relatedness and virulence phenotypic characteristics in vitro.

Study Design A total of 24 neonates with E. coli bacteremia were identified prospectively in a tertiary-care hospital. Clinical and antibiotic resistance data were investigated. The E. coli isolates were analyzed by multilocus sequence typing (MLST); the presence of the K1 capsule and their ability to invade intestinal epithelial cells were also assessed.

Results Most newborns were very low birth weight infants. Overall, 75% of the isolates were ampicillin resistant and 17% were gentamicin and tobramycin nonsusceptible. MLST determined sequence types 95 and 131 (ST95 and ST131) predominated, with ST131 becoming significantly more prevalent recently. The K1 capsule was present in 50% of the isolates. ST131 isolates and those producing bacteremia in newborns younger than 7 days showed a highly invasive phenotype.

Conclusion Resistance to antibiotics currently used empirically to treat newborns is present in bacteremia-producing E. coli. Clonal spread among newborns of multidrug-resistant E. coli is possible; therefore, continued surveillance is needed. Identification of additional virulence factors associated with increased invasion in neonatal E. coli strains is important and further studies are warranted.

Keywords

neonatal sepsis - Escherichia coli - antibiotic resistance - intestinal invasion

Full Text

References

References
1 Cordero L, Rau R, Taylor D, Ayers LW. Enteric gram-negative bacilli bloodstream infections: 17 years' experience in a neonatal intensive care unit. Am J Infect Control 2004; 32 (4) 189-195
2 Schrag SJ, Hadler JL, Arnold KE, Martell-Cleary P, Reingold A, Schuchat A. Risk factors for invasive, early-onset Escherichia coli infections in the era of widespread intrapartum antibiotic use. Pediatrics 2006; 118 (2) 570-576
3 Stoll BJ, Hansen N, Fanaroff AA , et al. Changes in pathogens causing early-onset sepsis in very-low-birth-weight infants. N Engl J Med 2002; 347 (4) 240-247
4 Bizzarro MJ, Dembry LM, Baltimore RS, Gallagher PG. Changing patterns in neonatal Escherichia coli sepsis and ampicillin resistance in the era of intrapartum antibiotic prophylaxis. Pediatrics 2008; 121 (4) 689-696
5 Stoll BJ, Hansen NI, Sánchez PJ , et al; Eunice Kennedy Shriver National Institute of Child Health and Human Development Neonatal Research Network. Early onset neonatal sepsis: the burden of group B Streptococcal and E. coli disease continues. Pediatrics 2011; 127 (5) 817-826
6 Cotten CM, Smith PB. Duration of empirical antibiotic therapy for infants suspected of early-onset sepsis. Curr Opin Pediatr 2013; 25 (2) 167-171
7 Clark RH, Bloom BT, Spitzer AR, Gerstmann DR. Empiric use of ampicillin and cefotaxime, compared with ampicillin and gentamicin, for neonates at risk for sepsis is associated with an increased risk of neonatal death. Pediatrics 2006; 117 (1) 67-74
8 Moore MR, Schrag SJ, Schuchat A. Effects of intrapartum antimicrobial prophylaxis for prevention of group-B-streptococcal disease on the incidence and ecology of early-onset neonatal sepsis. Lancet Infect Dis 2003; 3 (4) 201-213
9 Kim KS. Pathogenesis of bacterial meningitis: from bacteraemia to neuronal injury. Nat Rev Neurosci 2003; 4 (5) 376-385
10 Kim KS. Mechanisms of microbial traversal of the blood-brain barrier. Nat Rev Microbiol 2008; 6 (8) 625-634
11 Huang SH, He L, Zhou Y, Wu CH, Jong A. Lactobacillus rhamnosus GG suppresses meningitic E. coli K1 penetration across human intestinal epithelial cells in vitro and protects neonatal rats against experimental hematogenous meningitis. Int J Microbiol 2009; 2009: 647862
12 Burns JL, Griffith A, Barry JJ, Jonas M, Chi EY. Transcytosis of gastrointestinal epithelial cells by Escherichia coli K1. Pediatr Res 2001; 49 (1) 30-37
13 Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically; Approved Standard. M07–A9. Vol 32. No. 2. Clinical and Laboratory Standards Institute; 2012
14 Achtman M, Mercer A, Kusecek B , et al. Six widespread bacterial clones among Escherichia coli K1 isolates. Infect Immun 1983; 39 (1) 315-335
15 Wirth T, Falush D, Lan R , et al. Sex and virulence in Escherichia coli: an evolutionary perspective. Mol Microbiol 2006; 60 (5) 1136-1151
16 Sarff LD, McCracken GH, Schiffer MS , et al. Epidemiology of Escherichia coli K1 in healthy and diseased newborns. Lancet 1975; 1 (7916) 1099-1104
17 Hyde TB, Hilger TM, Reingold A, Farley MM, O'Brien KL, Schuchat A ; Active Bacterial Core surveillance (ABCs) of the Emerging Infections Program Network. Trends in incidence and antimicrobial resistance of early-onset sepsis: population-based surveillance in San Francisco and Atlanta. Pediatrics 2002; 110 (4) 690-695
18 Shane AL, Stoll BJ. Recent developments and current issues in the epidemiology, diagnosis, and management of bacterial and fungal neonatal sepsis. Am J Perinatol 2013; 30 (2) 131-141
19 Weston EJ, Pondo T, Lewis MM , et al. The burden of invasive early-onset neonatal sepsis in the United States, 2005-2008. Pediatr Infect Dis J 2011; 30 (11) 937-941
20 Hornik CP, Fort P, Clark RH , et al. Early and late onset sepsis in very-low-birth-weight infants from a large group of neonatal intensive care units. Early Hum Dev 2012; 88 (Suppl. 02) S69-S74
21 Puopolo KM, Eichenwald EC. No change in the incidence of ampicillin-resistant, neonatal, early-onset sepsis over 18 years. Pediatrics 2010; 125 (5) e1031-e1038
22 Stoll BJ, Hansen N, Fanaroff AA , et al. Late-onset sepsis in very low birth weight neonates: the experience of the NICHD Neonatal Research Network. Pediatrics 2002; 110 (2 Pt 1) 285-291
23 Stoll BJ, Hansen NI, Higgins RD , et al; National Institute of Child Health and Human Development. Very low birth weight preterm infants with early onset neonatal sepsis: the predominance of gram-negative infections continues in the National Institute of Child Health and Human Development Neonatal Research Network, 2002-2003. Pediatr Infect Dis J 2005; 24 (7) 635-639
24 Fanaroff AA, Stoll BJ, Wright LL , et al; NICHD Neonatal Research Network. Trends in neonatal morbidity and mortality for very low birthweight infants. Am J Obstet Gynecol 2007; 196 (2) 147, e1-e8
25 Vergnano S, Sharland M, Kazembe P, Mwansambo C, Heath PT. Neonatal sepsis: an international perspective. Arch Dis Child Fetal Neonatal Ed 2005; 90 (3) F220-F224
26 Friedman S, Shah V, Ohlsson A, Matlow AG. Neonatal Escherichia coli infections: concerns regarding resistance to current therapy. Acta Paediatr 2000; 89 (6) 686-689
27 Alarcon A, Peña P, Salas S, Sancha M, Omeñaca F. Neonatal early onset Escherichia coli sepsis: trends in incidence and antimicrobial resistance in the era of intrapartum antimicrobial prophylaxis. Pediatr Infect Dis J 2004; 23 (4) 295-299
28 Polin RA ; Committee on Fetus and Newborn. Management of neonates with suspected or proven early-onset bacterial sepsis. Pediatrics 2012; 129 (5) 1006-1015
29 Watt S, Lanotte P, Mereghetti L, Moulin-Schouleur M, Picard B, Quentin R. Escherichia coli strains from pregnant women and neonates: intraspecies genetic distribution and prevalence of virulence factors. J Clin Microbiol 2003; 41 (5) 1929-1935
30 Mahjoub-Messai F, Bidet P, Caro V , et al. Escherichia coli isolates causing bacteremia via gut translocation and urinary tract infection in young infants exhibit different virulence genotypes. J Infect Dis 2011; 203 (12) 1844-1849
31 Weissman SJ, Chattopadhyay S, Aprikian P , et al. Clonal analysis reveals high rate of structural mutations in fimbrial adhesins of extraintestinal pathogenic Escherichia coli . Mol Microbiol 2006; 59 (3) 975-988
32 Johnson JR, Johnston B, Clabots C, Kuskowski MA, Castanheira M. Escherichia coli sequence type ST131 as the major cause of serious multidrug-resistant E. coli infections in the United States. Clin Infect Dis 2010; 51 (3) 286-294
33 Johnson JR, Anderson JT, Clabots C, Johnston B, Cooperstock M. Within-household sharing of a fluoroquinolone-resistant Escherichia coli sequence type ST131 strain causing pediatric osteoarticular infection. Pediatr Infect Dis J 2010; 29 (5) 473-475
34 Chandramohan L, Revell PA. Prevalence and molecular characterization of extended-spectrum-β-lactamase-producing Enterobacteriaceae in a pediatric patient population. Antimicrob Agents Chemother 2012; 56 (9) 4765-4770
35 Adams-Sapper S, Diep BA, Perdreau-Remington F, Riley LW. Clonal composition and community clustering of drug-susceptible and -resistant Escherichia coli isolates from bloodstream infections. Antimicrob Agents Chemother 2013; 57 (1) 490-497
36 Weissman SJ, Johnson JR, Tchesnokova V , et al. High-resolution two-locus clonal typing of extraintestinal pathogenic Escherichia coli. Appl Environ Microbiol 2012; 78 (5) 1353-1360
37 Robbins JB, McCracken Jr GH, Gotschlich EC, Ørskov F, Ørskov I, Hanson LA. Escherichia coli K1 capsular polysaccharide associated with neonatal meningitis. N Engl J Med 1974; 290 (22) 1216-1220
38 Kim KJ, Elliott SJ, Di Cello F, Stins MF, Kim KS. The K1 capsule modulates trafficking of E. coli-containing vacuoles and enhances intracellular bacterial survival in human brain microvascular endothelial cells. Cell Microbiol 2003; 5 (4) 245-252
39 Rogers BA, Sidjabat HE, Paterson DL. Escherichia coli O25b-ST131: a pandemic, multiresistant, community-associated strain. J Antimicrob Chemother 2011; 66 (1) 1-14
40 Huang SH, Wan ZS, Chen YH, Jong AY, Kim KS. Further characterization of Escherichia coli brain microvascular endothelial cell invasion gene ibeA by deletion, complementation, and protein expression. J Infect Dis 2001; 183 (7) 1071-1078
41 Mora A, Herrera A, Mamani R , et al. Recent emergence of clonal group O25b:K1:H4-B2-ST131 ibeA strains among Escherichia coli poultry isolates, including CTX-M-9-producing strains, and comparison with clinical human isolates. Appl Environ Microbiol 2010; 76 (21) 6991-6997
42 Vejborg RM, Friis C, Hancock V, Schembri MA, Klemm P. A virulent parent with probiotic progeny: comparative genomics of Escherichia coli strains CFT073, Nissle 1917 and ABU 83972. Mol Genet Genomics 2010; 283 (5) 469-484
43 Meier C, Oelschlaeger TA, Merkert H, Korhonen TK, Hacker J. Ability of Escherichia coli isolates that cause meningitis in newborns to invade epithelial and endothelial cells. Infect Immun 1996; 64 (7) 2391-2399
44 Mukhopadhyay S, Puopolo KM. Risk assessment in neonatal early onset sepsis. Semin Perinatol 2012; 36 (6) 408-415
45 Graham III PL, Begg MD, Larson E, Della-Latta P, Allen A, Saiman L. Risk factors for late onset gram-negative sepsis in low birth weight infants hospitalized in the neonatal intensive care unit. Pediatr Infect Dis J 2006; 25 (2) 113-117
46 Shakir SM, Goldbeck JM, Morton DJ, Whitby PW, Chavez-Bueno S. Signature-Tagged Mutagenesis Identifies Escherichia coli Strain RS218 Genes Involved in Invasion of Intestinal Epithelial Cells (Abstract 692). Presented at the 112th General Meeting of the American Society for Microbiology. San Francisco, CA; June 2012