Intracellular and extracellular antimicrobial resistance genes in the sludge of livestock waste management structures

Yuping Zhang, Daniel D Snow, David Parker, Zhi Zhou, Xu Li

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

The sludge compartment in livestock waste management structures is a potential hotbed for the emergence and proliferation of antimicrobial resistance among bacteria. Little is known about the distribution of antimicrobial resistance genes (ARGs) between the intracellular and extracellular DNA pools in the sludge. The overall objective of this study was to assess the significance of extracellular ARGs to the total ARGs in the sludge of livestock waste management structures. In this study, sludge samples were collected from four cattle manure storage ponds and three swine waste treatment lagoons and analyzed for genetic indicators of resistance. Intracellular DNA (iDNA) and extracellular DNA (eDNA) in the sludge were separately extracted using an optimized protocol. ARGs [sul(I), sul(II), tet(O), tet(Q), and tet(X)] in both the iDNA and eDNA extracts were quantified using quantitative polymerase chain reaction (qPCR), and antimicrobials, including sulfonamides and tetracyclines, were measured using liquid chromatography tandem mass spectrometry. Results showed that eDNA constituted less than 1.5% of the total DNA in sludge. All ARGs tested were detected in nearly all eDNA and iDNA samples. Furthermore, every gram of dry sludge contained from 1.7 × 103 to 4.2 × 108 copies of extracellular ARG and from 3.2 × 107 to 3.2 × 1010 copies of intracellular ARG. Chlortetracycline concentrations ranged between 187 and 2674 μg/g of sludge wet weight (ww), while sulfonamide concentrations were lower than 6.3 μg/g of sludge ww. The detection of ARGs in eDNA extracts suggests that transformation is a potential mechanism in ARG proliferation in the sludge of livestock waste management structures.

Original languageEnglish (US)
Pages (from-to)10206-10213
Number of pages8
JournalEnvironmental Science and Technology
Volume47
Issue number18
DOIs
StatePublished - Sep 17 2013

Fingerprint

Waste management
Agriculture
waste management
livestock
Genes
sludge
DNA
gene
Sulfonamides
Chlortetracycline
Tetracyclines
Waste treatment
Manures
Polymerase chain reaction
Liquid chromatography
waste treatment
Ponds
polymerase chain reaction
Mass spectrometry
liquid chromatography

ASJC Scopus subject areas

  • Chemistry(all)
  • Environmental Chemistry

Cite this

Intracellular and extracellular antimicrobial resistance genes in the sludge of livestock waste management structures. / Zhang, Yuping; Snow, Daniel D; Parker, David; Zhou, Zhi; Li, Xu.

In: Environmental Science and Technology, Vol. 47, No. 18, 17.09.2013, p. 10206-10213.

Research output: Contribution to journalArticle

@article{13ba2b9e7051402fa20742b7ea73c3cc,
title = "Intracellular and extracellular antimicrobial resistance genes in the sludge of livestock waste management structures",
abstract = "The sludge compartment in livestock waste management structures is a potential hotbed for the emergence and proliferation of antimicrobial resistance among bacteria. Little is known about the distribution of antimicrobial resistance genes (ARGs) between the intracellular and extracellular DNA pools in the sludge. The overall objective of this study was to assess the significance of extracellular ARGs to the total ARGs in the sludge of livestock waste management structures. In this study, sludge samples were collected from four cattle manure storage ponds and three swine waste treatment lagoons and analyzed for genetic indicators of resistance. Intracellular DNA (iDNA) and extracellular DNA (eDNA) in the sludge were separately extracted using an optimized protocol. ARGs [sul(I), sul(II), tet(O), tet(Q), and tet(X)] in both the iDNA and eDNA extracts were quantified using quantitative polymerase chain reaction (qPCR), and antimicrobials, including sulfonamides and tetracyclines, were measured using liquid chromatography tandem mass spectrometry. Results showed that eDNA constituted less than 1.5{\%} of the total DNA in sludge. All ARGs tested were detected in nearly all eDNA and iDNA samples. Furthermore, every gram of dry sludge contained from 1.7 × 103 to 4.2 × 108 copies of extracellular ARG and from 3.2 × 107 to 3.2 × 1010 copies of intracellular ARG. Chlortetracycline concentrations ranged between 187 and 2674 μg/g of sludge wet weight (ww), while sulfonamide concentrations were lower than 6.3 μg/g of sludge ww. The detection of ARGs in eDNA extracts suggests that transformation is a potential mechanism in ARG proliferation in the sludge of livestock waste management structures.",
author = "Yuping Zhang and Snow, {Daniel D} and David Parker and Zhi Zhou and Xu Li",
year = "2013",
month = "9",
day = "17",
doi = "10.1021/es401964s",
language = "English (US)",
volume = "47",
pages = "10206--10213",
journal = "Environmental Science & Technology",
issn = "0013-936X",
publisher = "American Chemical Society",
number = "18",

}

TY - JOUR

T1 - Intracellular and extracellular antimicrobial resistance genes in the sludge of livestock waste management structures

AU - Zhang, Yuping

AU - Snow, Daniel D

AU - Parker, David

AU - Zhou, Zhi

AU - Li, Xu

PY - 2013/9/17

Y1 - 2013/9/17

N2 - The sludge compartment in livestock waste management structures is a potential hotbed for the emergence and proliferation of antimicrobial resistance among bacteria. Little is known about the distribution of antimicrobial resistance genes (ARGs) between the intracellular and extracellular DNA pools in the sludge. The overall objective of this study was to assess the significance of extracellular ARGs to the total ARGs in the sludge of livestock waste management structures. In this study, sludge samples were collected from four cattle manure storage ponds and three swine waste treatment lagoons and analyzed for genetic indicators of resistance. Intracellular DNA (iDNA) and extracellular DNA (eDNA) in the sludge were separately extracted using an optimized protocol. ARGs [sul(I), sul(II), tet(O), tet(Q), and tet(X)] in both the iDNA and eDNA extracts were quantified using quantitative polymerase chain reaction (qPCR), and antimicrobials, including sulfonamides and tetracyclines, were measured using liquid chromatography tandem mass spectrometry. Results showed that eDNA constituted less than 1.5% of the total DNA in sludge. All ARGs tested were detected in nearly all eDNA and iDNA samples. Furthermore, every gram of dry sludge contained from 1.7 × 103 to 4.2 × 108 copies of extracellular ARG and from 3.2 × 107 to 3.2 × 1010 copies of intracellular ARG. Chlortetracycline concentrations ranged between 187 and 2674 μg/g of sludge wet weight (ww), while sulfonamide concentrations were lower than 6.3 μg/g of sludge ww. The detection of ARGs in eDNA extracts suggests that transformation is a potential mechanism in ARG proliferation in the sludge of livestock waste management structures.

AB - The sludge compartment in livestock waste management structures is a potential hotbed for the emergence and proliferation of antimicrobial resistance among bacteria. Little is known about the distribution of antimicrobial resistance genes (ARGs) between the intracellular and extracellular DNA pools in the sludge. The overall objective of this study was to assess the significance of extracellular ARGs to the total ARGs in the sludge of livestock waste management structures. In this study, sludge samples were collected from four cattle manure storage ponds and three swine waste treatment lagoons and analyzed for genetic indicators of resistance. Intracellular DNA (iDNA) and extracellular DNA (eDNA) in the sludge were separately extracted using an optimized protocol. ARGs [sul(I), sul(II), tet(O), tet(Q), and tet(X)] in both the iDNA and eDNA extracts were quantified using quantitative polymerase chain reaction (qPCR), and antimicrobials, including sulfonamides and tetracyclines, were measured using liquid chromatography tandem mass spectrometry. Results showed that eDNA constituted less than 1.5% of the total DNA in sludge. All ARGs tested were detected in nearly all eDNA and iDNA samples. Furthermore, every gram of dry sludge contained from 1.7 × 103 to 4.2 × 108 copies of extracellular ARG and from 3.2 × 107 to 3.2 × 1010 copies of intracellular ARG. Chlortetracycline concentrations ranged between 187 and 2674 μg/g of sludge wet weight (ww), while sulfonamide concentrations were lower than 6.3 μg/g of sludge ww. The detection of ARGs in eDNA extracts suggests that transformation is a potential mechanism in ARG proliferation in the sludge of livestock waste management structures.

UR - http://www.scopus.com/inward/record.url?scp=84884277543&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84884277543&partnerID=8YFLogxK

U2 - 10.1021/es401964s

DO - 10.1021/es401964s

M3 - Article

VL - 47

SP - 10206

EP - 10213

JO - Environmental Science & Technology

JF - Environmental Science & Technology

SN - 0013-936X

IS - 18

ER -