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High frequencies of antibiotic resistance genes in infants’ meconium and early fecal samples

Published online by Cambridge University Press:  10 September 2015

M. J. Gosalbes
Affiliation:
Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO), Unitat Mixta d’Investigació en Genòmica i Salut, FISABIO-Salut Pública/Institut Cavanilles de Biodiversitat i Biologia Evolutiva, Valencia, Spain CIBER en Epidemiología y Salud Pública (CIBEResp), Madrid, Spain
Y. Vallès
Affiliation:
Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO), Unitat Mixta d’Investigació en Genòmica i Salut, FISABIO-Salut Pública/Institut Cavanilles de Biodiversitat i Biologia Evolutiva, Valencia, Spain
N. Jiménez-Hernández
Affiliation:
Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO), Unitat Mixta d’Investigació en Genòmica i Salut, FISABIO-Salut Pública/Institut Cavanilles de Biodiversitat i Biologia Evolutiva, Valencia, Spain
C. Balle
Affiliation:
Department of Biology, Section of Microbiology, University of Copenhagen, Copenhagen, Denmark
P. Riva
Affiliation:
Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO), Unitat Mixta d’Investigació en Genòmica i Salut, FISABIO-Salut Pública/Institut Cavanilles de Biodiversitat i Biologia Evolutiva, Valencia, Spain
S. Miravet-Verde
Affiliation:
Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO), Unitat Mixta d’Investigació en Genòmica i Salut, FISABIO-Salut Pública/Institut Cavanilles de Biodiversitat i Biologia Evolutiva, Valencia, Spain
L. E. de Vries
Affiliation:
Department of Biology, Section of Microbiology, University of Copenhagen, Copenhagen, Denmark Department of Technology, Metropolitan University College,Copenhagen, Denmark
S. Llop
Affiliation:
CIBER en Epidemiología y Salud Pública (CIBEResp), Madrid, Spain FISABIO-UJI-University of Valencia Epidemiology and Environmental Health Unit of Research, Valencia, Spain
Y. Agersø
Affiliation:
National Food Institute, Technical University of Denmark, Lyngby, Denmark
S. J. Sørensen
Affiliation:
Department of Biology, Section of Microbiology, University of Copenhagen, Copenhagen, Denmark
F. Ballester
Affiliation:
CIBER en Epidemiología y Salud Pública (CIBEResp), Madrid, Spain FISABIO-UJI-University of Valencia Epidemiology and Environmental Health Unit of Research, Valencia, Spain
M. P. Francino*
Affiliation:
Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO), Unitat Mixta d’Investigació en Genòmica i Salut, FISABIO-Salut Pública/Institut Cavanilles de Biodiversitat i Biologia Evolutiva, Valencia, Spain CIBER en Epidemiología y Salud Pública (CIBEResp), Madrid, Spain School of Natural Sciences, University of California Merced, Merced, CA, USA
*
*Address for correspondence: M. P. Francino, Unitat Mixta d’Investigació en Genòmica i Salut, FISABIO-Salut Pública, Ave. Catalunya 21, Valencia 46020, Spain. (Email [email protected])

Abstract

The gastrointestinal tract (GIT) microbiota has been identified as an important reservoir of antibiotic resistance genes (ARGs) that can be horizontally transferred to pathogenic species. Maternal GIT microbes can be transmitted to the offspring, and recent work indicates that such transfer starts before birth. We have used culture-independent genetic screenings to explore whether ARGs are already present in the meconium accumulated in the GIT during fetal life and in feces of 1-week-old infants. We have analyzed resistance to β-lactam antibiotics (BLr) and tetracycline (Tcr), screening for a variety of genes conferring each. To evaluate whether ARGs could have been inherited by maternal transmission, we have screened perinatal fecal samples of the 1-week-old babies’ mothers, as well as a mother–infant series including meconium, fecal samples collected through the infant’s 1st year, maternal fecal samples and colostrum. Our results reveal a high prevalence of BLr and Tcr in both meconium and early fecal samples, implying that the GIT resistance reservoir starts to accumulate even before birth. We show that ARGs present in the mother may reach the meconium and colostrum and establish in the infant GIT, but also that some ARGs were likely acquired from other sources. Alarmingly, we identified in both meconium and 1-week-olds’ samples a particularly elevated prevalence of mecA (>45%), six-fold higher than that detected in the mothers. The mecA gene confers BLr to methicillin-resistant Staphylococcus aureus, and although its detection does not imply the presence of this pathogen, it does implicate the young infant’s GIT as a noteworthy reservoir of this gene.

Type
Original Article
Copyright
© Cambridge University Press and the International Society for Developmental Origins of Health and Disease 2015 

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References

1. Salyers, AA, Gupta, A, Wang, Y. Human intestinal bacteria as reservoirs for antibiotic resistance genes. Trends Microbiol. 2004; 12, 412416.Google Scholar
2. Seville, LA, Patterson, AJ, Scott, KP, et al. Distribution of tetracycline and erythromycin resistance genes among human oral and fecal metagenomic DNA. Microb Drug Resist. 2009; 15, 159166.Google Scholar
3. Sommer, MO, Dantas, G, Church, GM. Functional characterization of the antibiotic resistance reservoir in the human microflora. Science. 2009; 325, 11281131.CrossRefGoogle Scholar
4. Moore, AM, Patel, S, Forsberg, KJ, et al. Pediatric fecal microbiota harbor diverse and novel antibiotic resistance genes. PLoS One. 2013; 8, e78822.Google Scholar
5. Fouhy, F, Ogilvie, LA, Jones, BV, et al. Identification of aminoglycoside and beta-lactam resistance genes from within an infant gut functional metagenomic library. PLoS One. 2014; 9, e108016.Google Scholar
6. Lu, N, Hu, Y, Zhu, L, et al. DNA microarray analysis reveals that antibiotic resistance-gene diversity in human gut microbiota is age related. Sci Rep. 2014; 4, 4302.Google Scholar
7. Hu, Y, Yang, X, Lu, N, Zhu, B. The abundance of antibiotic resistance genes in human guts has correlation to the consumption of antibiotics in animal. Gut Microbes. 2014; 5, 245249.Google Scholar
8. Gueimonde, M, Salminen, S, Isolauri, E. Presence of specific antibiotic (tet) resistance genes in infant faecal microbiota. FEMS Immunol Med Microbiol. 2006; 48, 2125.Google Scholar
9. Mitsou, EK, Kirtzalidou, E, Pramateftaki, P, Kyriacou, A. Antibiotic resistance in faecal microbiota of greek healthy infants. Benef Microbes. 2010; 1, 297306.Google Scholar
10. de Vries, LE, Valles, Y, Agerso, Y, et al. The gut as reservoir of antibiotic resistance: microbial diversity of tetracycline resistance in mother and infant. PLoS One. 2011; 6, e21644.Google Scholar
11. Zhang, L, Kinkelaar, D, Huang, Y, et al.. Acquired antibiotic resistance: are we born with it? Appl Environ Microbiol. 2011; 77, 71347141.Google Scholar
12. Alicea-Serrano, AM, Contreras, M, Magris, M, Hidalgo, G, Dominguez-Bello, MG. Tetracycline resistance genes acquired at birth. Arch Microbiol. 2013; 195, 447451.Google Scholar
13. Collado, MC, Isolauri, E, Laitinen, K, Salminen, S. Distinct composition of gut microbiota during pregnancy in overweight and normal-weight women. Am J Clin Nutr. 2008; 88, 894899.Google Scholar
14. Gilbert, SF. A holobiont birth narrative: the epigenetic transmission of the human microbiome. Front Genet. 2014; 5, 282.Google Scholar
15. Koren, O, Goodrich, JK, Cullender, TC, et al. Host remodeling of the gut microbiome and metabolic changes during pregnancy. Cell. 2012; 150, 470480.Google Scholar
16. Berg, R. Bacterial translocation from the gastrintestinal tract. Trends Microbiol. 1995; 3, 149154.Google Scholar
17. Cani, PD, Delzenne, NM. The gut microbiome as therapeutic target. Pharmacol Ther. 2011; 130, 202212.Google Scholar
18. Francino, MP. Early development of the gut microbiota and immune health. Pathogens. 2014; 3, 769790.CrossRefGoogle Scholar
19. Gosalbes, MJ, Llop, S, Valles, Y, et al. Meconium microbiota types dominated by lactic acid or enteric bacteria are differentially associated with maternal eczema and respiratory problems in infants. Clin Exp Allergy. 2013; 43, 198211.Google Scholar
20. Valles, Y, Artacho, A, Pascual-Garcia, A, et al. Microbial succession in the gut: directional trends of taxonomic and functional change in a birth cohort of Spanish infants. PLoS Genet. 2014; 10, e1004406.Google Scholar
21. Perez, PF, Dore, J, Leclerc, M, et al.. Bacterial imprinting of the neonatal immune system: lessons from maternal cells? Pediatrics. 2007; 119, e724e732.Google Scholar
22. Donnet-Hughes, A, Perez, PF, Dore, J, et al. Potential role of the intestinal microbiota of the mother in neonatal immune education. Proc Nutr Soc. 2010; 69, 407415.Google Scholar
23. Amar, J, Chabo, C, Waget, A, et al. Intestinal mucosal adherence and translocation of commensal bacteria at the early onset of type 2 diabetes: molecular mechanisms and probiotic treatment. EMBO Mol Med. 2011; 3, 559572.Google Scholar
24. McGovern, N, Chan, JK, Ginhoux, F. Dendritic cells in humans – from fetus to adult. Int Immunol. 2015; 27, 6572.Google Scholar
25. DiGiulio, DB, Romero, R, Amogan, HP, et al. Microbial prevalence, diversity and abundance in amniotic fluid during preterm labor: a molecular and culture-based investigation. PLoS One. 2008; 3, e3056.Google Scholar
26. DiGiulio, DB, Romero, R, Kusanovic, JP, et al. Prevalence and diversity of microbes in the amniotic fluid, the fetal inflammatory response, and pregnancy outcome in women with preterm pre-labor rupture of membranes. Am J Reprod Immunol. 2010; 64, 3857.Google Scholar
27. Dong, Y, St Clair, PJ, Ramzy, I, Kagan-Hallet, KS, Gibbs, RS. A microbiologic and clinical study of placental inflammation at term. Obstet Gynecol. 1987; 70, 175182.Google Scholar
28. Lewis, JF, Johnson, P, Miller, P. Evaluation of amniotic fluid for aerobic and anaerobic bacteria. Am J Clin Pathol. 1976; 65, 5863.Google Scholar
29. Gibbs, RS, Blanco, JD, St Clair, PJ, Castaneda, YS. Quantitative bacteriology of amniotic fluid from women with clinical intraamniotic infection at term. J Infect Dis. 1982; 145, 18.Google Scholar
30. Romero, R, Mazor, M, Morrotti, R, et al. Infection and labor. Vii. Microbial invasion of the amniotic cavity in spontaneous rupture of membranes at term. Am J Obstet Gynecol. 1992; 166, 129133.Google Scholar
31. Romero, R, Sirtori, M, Oyarzun, E, et al. Infection and labor. V. Prevalence, microbiology, and clinical significance of intraamniotic infection in women with preterm labor and intact membranes. Am J Obstet Gynecol. 1989; 161, 817824.Google Scholar
32. Bearfield, C, Davenport, ES, Sivapathasundaram, V, Allaker, RP. Possible association between amniotic fluid micro-organism infection and microflora in the mouth. BJOG. 2002; 109, 527533.Google Scholar
33. Steel, JH, Malatos, S, Kennea, N, et al.. Bacteria and inflammatory cells in fetal membranes do not always cause preterm labor. Pediatr Res. 2005; 57, 404411.Google Scholar
34. Jimenez, E, Fernandez, L, Marin, ML, et al.. Isolation of commensal bacteria from umbilical cord blood of healthy neonates born by cesarean section. Curr Microbiol. 2005; 51, 270274.Google Scholar
35. Roos, PJ, Malan, AF, Woods, DL, et al. The bacteriological environment of preterm infants. S Afr Med J. 1980; 57, 347350.Google Scholar
36. Stout, MJ, Conlon, B, Landeau, M, et al. Identification of intracellular bacteria in the basal plate of the human placenta in term and preterm gestations. Am J Obstet Gynecol 2013; 208, 226 e221226 e227.Google Scholar
37. Aagaard, K, Ma, J, Antony, KM, et al. The placenta harbors a unique microbiome. Sci Transl Med 2014; 6, 237ra265.Google Scholar
38. Jimenez, E, Marin, ML, Martin, R, et al. Is meconium from healthy newborns actually sterile? Res Microbiol. 2008; 159, 187193.Google Scholar
39. Koenig, JE, Spor, A, Scalfone, N, et al. Succession of microbial consortia in the developing infant gut microbiome. Proc Natl Acad Sci U S A. 2011; 108(Suppl. 1), 45784585.Google Scholar
40. Moles, L, Gomez, M, Heilig, H, et al. Bacterial diversity in meconium of preterm neonates and evolution of their fecal microbiota during the first month of life. PLoS One. 2013; 8, e66986.Google Scholar
41. Dominguez-Bello, MG, Costello, EK, Contreras, M, et al. Delivery mode shapes the acquisition and structure of the initial microbiota across multiple body habitats in newborns. Proc Natl Acad Sci U S A. 2010; 107, 1197111975.Google Scholar
42. Hu, J, Nomura, Y, Bashir, A, et al. Diversified microbiota of meconium is affected by maternal diabetes status. PLoS One. 2013; 8, e78257.Google Scholar
43. Madan, JC, Salari, RC, Saxena, D, et al. Gut microbial colonisation in premature neonates predicts neonatal sepsis. Arch Dis Child Fetal Neonatal Ed. 2012; 97, F456F462.Google Scholar
44. Mshvildadze, M, Neu, J, Shuster, J, et al. Intestinal microbial ecology in premature infants assessed with non-culture-based techniques. J Pediatr. 2010; 156, 2025.Google Scholar
45. Valles, Y, Gosalbes, MJ, de Vries, LE, Abellan, JJ, Francino, MP. Metagenomics and development of the gut microbiota in infants. Clin Microbiol Infect. 2012; 18(Suppl. 4), 2126.Google Scholar
46. Cabrera-Rubio, R, Collado, MC, Laitinen, K, et al. The human milk microbiome changes over lactation and is shaped by maternal weight and mode of delivery. Am J Clin Nutr. 2012; 96, 544551.Google Scholar
47. Nichols, DA, Renslo, AR, Chen, Y. Fragment-based inhibitor discovery against beta-lactamase. Fut Med Chem. 2014; 6, 413427.CrossRefGoogle Scholar
48. Lowy, FD. Antimicrobial resistance: the example of Staphylococcus aureus . J Clin Invest. 2003; 111, 12651273.Google Scholar
49. Kazimierczak, KA, Scott, KP, Kelly, D, Aminov, RI. Tetracycline resistome of the organic pig gut. Appl Environ Microbiol. 2009; 75, 17171722.Google Scholar
50. Ribas-Fito, N, Ramon, R, Ballester, F, et al. Child health and the environment: the INMA Spanish Study. Paediatr Perinat Epidemiol. 2006; 20, 403410.Google Scholar
51. Ng, LK, Martin, I, Alfa, M, Mulvey, M. Multiplex pcr for the detection of tetracycline resistant genes. Mol Cell Probes. 2001; 15, 209215.Google Scholar
52. Clemente, JC, Pehrsson, EC, Blaser, MJ, et al. The microbiome of uncontacted amerindians. Sci Adv. 2015; 1, e1500183.Google Scholar
53. Agerso, Y, Aarestrup, FM, Pedersen, K, et al. Prevalence of extended-spectrum cephalosporinase (esc)-producing Escherichia coli in danish slaughter pigs and retail meat identified by selective enrichment and association with cephalosporin usage. J Antimicrob Chemother. 2012; 67, 582588.Google Scholar
54. Poirel, L, Walsh, TR, Cuvillier, V, Nordmann, P. Multiplex pcr for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis. 2011; 70, 119123.Google Scholar
55. Mendes, RE, Kiyota, KA, Monteiro, J, et al. Rapid detection and identification of metallo-β-lactamase-encoding genes by multiplex real-time PCR assay and melt curve analysis. J Clin Microbiol. 2007; 45, 544547.Google Scholar
56. Poulsen, AB, Skov, R, Pallesen, LV. Detection of methicillin resistance in coagulase-negative staphylococci and in staphylococci directly from simulated blood cultures using the evigene mrsa detection kit. J Antimicrob Chemother. 2003; 51, 419421.Google Scholar