tracking antibiotics and tetracycline resistance genes
TRANSCRIPT
LOGO
Tracking Antibiotics and Tetracycline Resistance Genes
through the Poultry Waste Composting Process – Lessons
Learned
Patricia Keen, PhD
Institute for Resources, Environment & Sustainability
Tracking Antibiotics through Poultry Compost Process
Outline
1. Project Introduction
2. Experiment design/methods
3. Results
4. Lessons learned
Tracking Antibiotics through Poultry Compost Process
Why are Antibiotics in the Environment a concern?
The surviving bacteria can resist the action of the antibiotic
The ‘fittest” bacteria survive
Bioactive compounds act on the organism
Prescription drugs Involuntary exposure
Antimicrobial Resistance
Natural Selection of Bacteria
Changes in Cellular Biochemistry
Exposure of Bacteria to Antibiotics
Antibiotics in an Agricultural Watershed
AMR in the Environment
Exchange genetic
determinants of
resistance
Transport in
Environment
Development of
Resistance
Tracking Antibiotics through Poultry Compost Process
Objectives
To refine LC ESI-MS/MS methodology for analyses of low level antibiotic concentrations in environmental samples.
To determine the concentration of antibiotics and tetracycline resistance genes in samples of waste from poultry production to track through the composting process.
To investigate whether monitoring antibiotic residues or antibiotic resistance genes could indicate potential for spread of antimicrobial resistance through environmental pathways.
To follow the persistence of four selected tetracycline resistance genes on a fertilized field
Tracking Antibiotics through Poultry Compost Process
Sumas Watershed
Headwaters in Whatcom County, USA
Flows across the international border
Drains into the Fraser River at the mouth of the Vedder Canal
Lower Fraser Valley, British Columbia
The Sumas River
Tracking Antibiotics through Poultry Compost Process
Study Design
Measure antibiotics/tetracycline
resistance genes compost samples
Monitor the analytes through
the composting process
Follow the tetracycline resistance
genes after application on field
To examine how
composting affects the
pathways by which
bacteria could develop
AMR via environmental
exposure
Tracking Antibiotics through Poultry Compost Process
Antibiotics Monitored 2004-2005 in the Sumas Watershed
Oxytetracycline Sulphamethazine Monensin
Chloroxytetracycline Sulphathiazole Trimethoprim
Democlocycline Sulphadimethoxine Ormetroprim
Doxycyline Sulphapyridine Narasin
Tetracycline Sulphaguanidine Salinmycin sodium
Tylosin Sulphamethizole Lasolicid sodium
Tilmicosin Sulphachloropyridazine Gentimycin
Virginiamycin Sulphamethoxazole Spectinomycin
Lincomycin Sulphamerizine Erythromycin
Tracking Antibiotics through Poultry Compost Process
Antibiotics Measured in the Sumas Watershed between 2-100 ppb
Oxytetracycline
Chloroxytetracycline
Democlocycline
Doxycyline
Tetracycline
Tylosin
Tilmicosin
Virginiamycin
Lincomycin
Sulphamethazine
Sulphathiazole
Sulphadimethoxine
Sulphapyridine
Sulphaguanidine
Sulphamethizole
Sulphachloropyridazine
Sulphamethoxazole
Sulphamerizine
Monensin
Trimethoprim
Ormetroprim
Narasin
Salinmycin sodium
Lasolicid sodium
Gentimycin
Spectinomycin
Erythromycin
Tracking Antibiotics through Poultry Compost Process
Antibiotic Residues Detected in Receiving Water and Soil – Sumas Watershed
doxycycline 3.7 – 11.1
democlocycline 3.8 – 27.0
oxytetracycline 3.1 – 57.9
chloroxytetracycline 3.9 – 37.2
tetracycline 3.2 – 9.1
Receiving Water (ppb)
(July 04 – April 05)
doxycycline n/d
democlocycline n/d
oxytetracycline 20 – 40
chloroxytetracycline n/d
tetracycline n/d
sulphamethizole 20 – 25
sulphathiazole 20 – 25
virginiamycin 20 – 40
lincomycin 20 – 25
erythromycin 20 – 25
Soil (ppb)
(April 05)
Total Tetracycline Resistance Genes in Compost & Soil
Tracking Antibiotics through Poultry Compost Process
Total tet Resistance Genes - Poultry Compost & Soils
1.00E+00
1.00E+01
1.00E+02
1.00E+03
1.00E+04
1.00E+05
1.00E+06
1.00E+07
1.00E+08
1.00E+09
Jan
25/0
4 ch
icke
n litte
r bef
ore
bird
s
June
2/0
4 co
mpo
st
July 2
3/04
com
post
Aug
20/
04 com
post
Sep
t 6/0
4 co
mpo
st
Sep
t PRE s
pray
soi
l
Sep
t 8/0
4 so
il
Sep
t 15/
04 s
oil
Sep
t 24/
04 s
oil
Oct 8
/04
soil
Oct
14/
04 s
oil
Oct 2
0/04
soi
l
Oct
29/
04 s
oil
Nov
4/0
4 so
ilDec
3/0
4 so
il
Dec
10/
04 soi
l
Jan
25/0
5 so
il
Sample date
Ab
un
da
nc
e C
op
ies
/mg
tet M
tet Q
tet O
tet W
Total Tcr and 16S rRNA genes
Tracking Antibiotics through Poultry Compost Process
1.00E+00
1.00E+01
1.00E+02
1.00E+03
1.00E+04
1.00E+05
1.00E+06
1.00E+07
1.00E+08
1.00E+09
1.00E+10
Jan
25/0
4 ch
icke
n litte
r bef
ore
bird
s
June
2/0
4 co
mpo
st
July 2
3/04
com
post
Aug
20/
04 com
post
Sep
t 6/0
4 co
mpo
st
Sep
t PRE s
pray
soi
l
Sep
t 8/0
4 so
il
Sep
t 15/
04 s
oil
Sep
t 24/
04 s
oil
Oct 8
/04
soil
Oct
14/
04 s
oil
Oct 2
0/04
soi
l
Oct
29/
04 s
oil
Nov
4/0
4 so
il
Dec
3/0
4 so
il
Dec
10/
04 soi
l
Jan
25/0
5 so
il
Sample Date
Co
pie
s/m
g
Total tet resistance genes
16S rRNA
Tracking Antibiotics through Poultry Compost Process
Prevalence of multi-drug resistance Salmonelle spp. isolates recovered from broiler chickens to 16 antimicrobials on the
NARMS panel (n=16)
Tracking Antibiotics through Poultry Compost Process
Prevalence of multi-drug resistance E.coli isolates recovered from broiler chickens to 16 antimicrobials on the NARMS
panel (n=16)
Tracking Antibiotics through Poultry Compost Process
Project Results - Summary
Some antibiotics require further method development for accurate and reliable results in compost/soil matrices (eg. Aminoglycocides)
Some antibiotic residues have been observed in ‘organic’ compost
Antibiotics present in the Sumas watershed coincide with local land use practices
There appears to be no correlation between tetracyclinesfound in the poultry compost samples and tetracycline resistance genes
Results appeared to be in line with observations of other global researchers
Lessons Learned
Measuring the antibiotic residues did not yield sufficient evidence to reliably monitor their presence or persistence
Analytical methodology required further refinement
Measuring selected tetracycline resistance genes allowed some observation of effects
Selecting an agricultural control site is extremely difficult –implications for experimental design
Some management practises can be recommended
Antibiotics in an Agricultural Watershed
Tracking Antibiotics through Poultry Compost Process
Project Collaborators
Health Santé
Canada Canada
Dr. Patricia Keen, University of British ColumbiaDr. Ken Hall, University of British ColumbiaDr. Bob Hancock, OC, University of British ColumbiaDr. Pierre Berubé, University of British ColumbiaDr. Julian Davies, OC, University of British Columbia
Dr. David Graham, University of KansasDr. Charles Knapp, University of Kansas
Dr. Merv Wetzstein, BC MALDr. Nancy deWith, BC MALDr. Heather Hannah, BC MALDr. Jane Pritchard, BC MAL
Dr. Dayue Shang, Health Canada Derek Smith, UBC Department of Chemistry