Antibiotic resistance associated with bacteria in irrigation water : a case study of irrigation ponds in Southern Sweden
The focus of this project was to characterize the occurrence of ESBL-‐producing bacteria in two irrigation water ponds in Southern Sweden. Samples were taken from two ponds nearby each other, from which analyses were made based on levels of community and ind...
| Autor principal: | |
|---|---|
| Formato: | H2 |
| Lenguaje: | Inglés sueco |
| Publicado: |
SLU/Plant Breeding and Biotechnology (until 121231)
2013
|
| Materias: |
| _version_ | 1855570818078081024 |
|---|---|
| author | Grudén, Maria |
| author_browse | Grudén, Maria |
| author_facet | Grudén, Maria |
| author_sort | Grudén, Maria |
| collection | Epsilon Archive for Student Projects |
| description | The
focus
of
this
project
was
to
characterize
the
occurrence
of
ESBL-‐producing
bacteria
in
two
irrigation
water
ponds
in
Southern
Sweden.
Samples
were
taken
from
two
ponds
nearby
each
other,
from
which
analyses
were
made
based
on
levels
of
community
and
individual
isolates.
Community
samples
were
used
for
attempts
for
characterization
of
resistance
patterns
in
irrigation
water
using
the
Omnilog
PM-‐plate
system.
The
growth
of
community
microbiota
in
the
presence
of
the
following
antibiotic
substances
was
assessed;
amikacin,
gentamicin,
kanamycin,
penicillin
G,
ampicillin,
piperacillin,
trimethoprim,
potassium
tellurite
and
ciprofloxacin.
Isolates
were
pure
cultured
using
semi-‐selective
media:
LB,
Rainbow
Agar,
mEnterococcus,
0.1
TSA
and
VRBD.
A
total
of
177
isolates
were
isolated
randomly.
Each
isolate
was
identified
using
the
Omnilog
GENIII-‐system
of
biochemical
characterization.
Further,
the
isolates
were
tested
on
Brilliance
ESBL-‐agar
for
their
ability
to
grow.
Out
of
the
total
number
of
isolates,
30
were
selected,
preferably
Enterobacteriaceae
or
Pseudomonas
spp.,
based
on
their
high
similarity
to
library
strain
during
the
identification
in
the
Omnilog-‐system.
These
were
further
characterized
using
Etest-‐sticks.
Three
selected
strains
were
picked
for
further
analyses
using
exogenous
isolation
for
attempts
to
simulate
horizontal
gene
transfer,
and
Omnilog
PM-‐panels
containing
various
antibiotics
of
interest.
Selections
of
samples
were
chosen
for
molecular
tests
targeting
plasmid-‐borne
resistance
genes
CTX-‐M1,
CTX-‐M2
and
SHV.
The
method
for
assessing
community-‐resistance
patterns
showed
ambiguous
data
that
was
difficult
to
interpret;
further
optimization
is
recommended.
For
all
tested
PM-‐plates,
generally
greatest
differences
between
substances
were
found
among
wells
with
the
strongest
concentration
of
antibiotic
substance.
A
total
of
175
isolates
displayed
ability
to
grow
on
ESBL-‐agar.
Etests
confirmed
ability
of
isolates
to
resist
many
β-‐lactam
antibiotics
and
many
were
clinically
resistant
to
certain
substances.
No
plasmids
were
transferred
during
exogenous
isolation.
During
molecular
tests,
a
few
matches
were
found
for
the
CTX-‐M1
gene.
Isolates
analyzed
with
Omnilog
PM-‐plates
showed
resistance
to
most
compounds
tested.
The
isolates
showed
greatest
resistance
toward
penicillin
G,
ampicillin,
amikacin,
kanamycin
and
trimethoprim.
In
the
study,
it
was
established
that
resistance
among
bacteria
was
frequently
occurring
in
the
irrigation
water,
and
β-‐lactamase
producing
bacteria
was
very
common.
Extended
studies
are
needed
to
assess
the
frequency-‐
and
occurrence
of
horizontal
gene
transfer
(HGF)
in
this
environment.
Although
some
potential
corresponding
data
is
found,
it
is
at
this
point
not
possible
to
determine
whether
the
resistances
are
due
to
antibiotics
used
in
veterinary
medicine,
human
medicine
or
one
additional
reason
is
travel
and
trade.
in
irrigation
water
using
the
Omnilog
PM-‐plate
system.
The
growth
of
community
microbiota
in
the
presence
of
the
following
antibiotic
substances
was
assessed;
amikacin,
gentamicin,
kanamycin,
penicillin
G,
ampicillin,
piperacillin,
trimethoprim,
potassium
tellurite
and
ciprofloxacin.
Isolates
were
pure
cultured
using
semi-‐selective
media:
LB,
Rainbow
Agar,
mEnterococcus,
0.1
TSA
and
VRBD.
A
total
of
177
isolates
were
isolated
randomly.
Each
isolate
was
identified
using
the
Omnilog
GENIII-‐system
of
biochemical
characterization.
Further,
the
isolates
were
tested
on
Brilliance
ESBL-‐agar
for
their
ability
to
grow.
Out
of
the
total
number
of
isolates,
30
were
selected,
preferably
Enterobacteriaceae
or
Pseudomonas
spp.,
based
on
their
high
similarity
to
library
strain
during
the
identification
in
the
Omnilog-‐system.
These
were
further
characterized
using
Etest-‐sticks.
Three
selected
strains
were
picked
for
further
analyses
using
exogenous
isolation
for
attempts
to
simulate
horizontal
gene
transfer,
and
Omnilog
PM-‐panels
containing
various
antibiotics
of
interest.
Selections
of
samples
were
chosen
for
molecular
tests
targeting
plasmid-‐borne
resistance
genes
CTX-‐M1,
CTX-‐M2
and
SHV.
The
method
for
assessing
community-‐resistance
patterns
showed
ambiguous
data
that
was
difficult
to
interpret;
further
optimization
is
recommended.
For
all
tested
PM-‐plates,
generally
greatest
differences
between
substances
were
found
among
wells
with
the
strongest
concentration
of
antibiotic
substance.
A
total
of
175
isolates
displayed
ability
to
grow
on
ESBL-‐agar.
Etests
confirmed
ability
of
isolates
to
resist
many
β-‐lactam
antibiotics
and
many
were
clinically
resistant
to
certain
substances.
No
plasmids
were
transferred
during
exogenous
isolation.
During
molecular
tests,
a
few
matches
were
found
for
the
CTX-‐M1
gene.
Isolates
analyzed
with
Omnilog
PM-‐plates
showed
resistance
to
most
compounds
tested.
The
isolates
showed
greatest
resistance
toward
penicillin
G,
ampicillin,
amikacin,
kanamycin
and
trimethoprim.
In
the
study,
it
was
established
that
resistance
among
bacteria
was
frequently
occurring
in
the
irrigation
water,
and
β-‐lactamase
producing
bacteria
was
very
common.
Extended
studies
are
needed
to
assess
the
frequency-‐
and
occurrence
of
horizontal
gene
transfer
(HGF)
in
this
environment.
Although
some
potential
corresponding
data
is
found,
it
is
at
this
point
not
possible
to
determine
whether
the
resistances
are
due
to
antibiotics
used
in
veterinary
medicine,
human
medicine
or
one
additional
reason
is
travel
and
trade. |
| format | H2 |
| id | RepoSLU5279 |
| institution | Swedish University of Agricultural Sciences |
| language | Inglés swe |
| publishDate | 2013 |
| publishDateSort | 2013 |
| publisher | SLU/Plant Breeding and Biotechnology (until 121231) |
| publisherStr | SLU/Plant Breeding and Biotechnology (until 121231) |
| record_format | eprints |
| spelling | RepoSLU52792015-10-21T14:27:21Z Antibiotic resistance associated with bacteria in irrigation water : a case study of irrigation ponds in Southern Sweden Antibiotikaresistens hos bakterier i bevattningsvatten : en fallstudie från två bevattningsdammar södra Sverige Grudén, Maria Animal husbandry Enterobacteriaceae ESBL Etest exogenous isolation irrigation water hygiene standards in water Water quality The focus of this project was to characterize the occurrence of ESBL-‐producing bacteria in two irrigation water ponds in Southern Sweden. Samples were taken from two ponds nearby each other, from which analyses were made based on levels of community and individual isolates. Community samples were used for attempts for characterization of resistance patterns in irrigation water using the Omnilog PM-‐plate system. The growth of community microbiota in the presence of the following antibiotic substances was assessed; amikacin, gentamicin, kanamycin, penicillin G, ampicillin, piperacillin, trimethoprim, potassium tellurite and ciprofloxacin. Isolates were pure cultured using semi-‐selective media: LB, Rainbow Agar, mEnterococcus, 0.1 TSA and VRBD. A total of 177 isolates were isolated randomly. Each isolate was identified using the Omnilog GENIII-‐system of biochemical characterization. Further, the isolates were tested on Brilliance ESBL-‐agar for their ability to grow. Out of the total number of isolates, 30 were selected, preferably Enterobacteriaceae or Pseudomonas spp., based on their high similarity to library strain during the identification in the Omnilog-‐system. These were further characterized using Etest-‐sticks. Three selected strains were picked for further analyses using exogenous isolation for attempts to simulate horizontal gene transfer, and Omnilog PM-‐panels containing various antibiotics of interest. Selections of samples were chosen for molecular tests targeting plasmid-‐borne resistance genes CTX-‐M1, CTX-‐M2 and SHV. The method for assessing community-‐resistance patterns showed ambiguous data that was difficult to interpret; further optimization is recommended. For all tested PM-‐plates, generally greatest differences between substances were found among wells with the strongest concentration of antibiotic substance. A total of 175 isolates displayed ability to grow on ESBL-‐agar. Etests confirmed ability of isolates to resist many β-‐lactam antibiotics and many were clinically resistant to certain substances. No plasmids were transferred during exogenous isolation. During molecular tests, a few matches were found for the CTX-‐M1 gene. Isolates analyzed with Omnilog PM-‐plates showed resistance to most compounds tested. The isolates showed greatest resistance toward penicillin G, ampicillin, amikacin, kanamycin and trimethoprim. In the study, it was established that resistance among bacteria was frequently occurring in the irrigation water, and β-‐lactamase producing bacteria was very common. Extended studies are needed to assess the frequency-‐ and occurrence of horizontal gene transfer (HGF) in this environment. Although some potential corresponding data is found, it is at this point not possible to determine whether the resistances are due to antibiotics used in veterinary medicine, human medicine or one additional reason is travel and trade. in irrigation water using the Omnilog PM-‐plate system. The growth of community microbiota in the presence of the following antibiotic substances was assessed; amikacin, gentamicin, kanamycin, penicillin G, ampicillin, piperacillin, trimethoprim, potassium tellurite and ciprofloxacin. Isolates were pure cultured using semi-‐selective media: LB, Rainbow Agar, mEnterococcus, 0.1 TSA and VRBD. A total of 177 isolates were isolated randomly. Each isolate was identified using the Omnilog GENIII-‐system of biochemical characterization. Further, the isolates were tested on Brilliance ESBL-‐agar for their ability to grow. Out of the total number of isolates, 30 were selected, preferably Enterobacteriaceae or Pseudomonas spp., based on their high similarity to library strain during the identification in the Omnilog-‐system. These were further characterized using Etest-‐sticks. Three selected strains were picked for further analyses using exogenous isolation for attempts to simulate horizontal gene transfer, and Omnilog PM-‐panels containing various antibiotics of interest. Selections of samples were chosen for molecular tests targeting plasmid-‐borne resistance genes CTX-‐M1, CTX-‐M2 and SHV. The method for assessing community-‐resistance patterns showed ambiguous data that was difficult to interpret; further optimization is recommended. For all tested PM-‐plates, generally greatest differences between substances were found among wells with the strongest concentration of antibiotic substance. A total of 175 isolates displayed ability to grow on ESBL-‐agar. Etests confirmed ability of isolates to resist many β-‐lactam antibiotics and many were clinically resistant to certain substances. No plasmids were transferred during exogenous isolation. During molecular tests, a few matches were found for the CTX-‐M1 gene. Isolates analyzed with Omnilog PM-‐plates showed resistance to most compounds tested. The isolates showed greatest resistance toward penicillin G, ampicillin, amikacin, kanamycin and trimethoprim. In the study, it was established that resistance among bacteria was frequently occurring in the irrigation water, and β-‐lactamase producing bacteria was very common. Extended studies are needed to assess the frequency-‐ and occurrence of horizontal gene transfer (HGF) in this environment. Although some potential corresponding data is found, it is at this point not possible to determine whether the resistances are due to antibiotics used in veterinary medicine, human medicine or one additional reason is travel and trade. Målet med detta projekt var att karaktärisera förekomsten av ESBL-‐producerande bakterier i bevattningsvatten i södra Sverige. Prover togs från två närbelägna bevattningsdammar, från vilka analyser gjordes på enskilda isolat och direkta prover. De prover som användes utan att selektera fram enskilda isolat användes för att testa-‐ och utvärdera en metod för karakterisering av resistensmönster i bevattningsvatten med hjälp av Omnilogs system med PM-‐plattor. I detta test bedömdes tillväxten av mikroorganismerna i närvaro av följande antibiotiska ämnen: amikacin, gentamicin, kanamycin, penicillin G, ampicillin, piperacillin, trimetoprim, kalium tellurit och ciprofloxacin. Ur samma prover isolerade enskilda isolat på följande medium: LB, Rainbow Agar, mEnterococcus, 0,1 TSA och VRBD. Totalt isolerades 177 stammar vilka plockades baserat på utseende på media, där heterogenitet i insamlat material var målet. Varje isolat identifierades med användning av Omnilog GENIII-‐system av biokemisk karakterisering. Vidare var isolaten testades på Brilliance ESBL-‐agar för deras förmåga att växa genom att producera betalaktamaser. Av det totala antalet isolat var valdes 30 stycken, företrädesvis Enterobacteriaceae eller Pseudomonas spp., baserat på likhet med modellorganismer under identifieringen i Omnilog-‐systemet. Dessa isolat karaktäriserades ytterligare med Etest-‐stickor. Utifrån dessa tester valdes tre stammar för ytterligare analyser där försök gjordes att överföra plasmider i en ansats att simulera horisontell genöverföring. Isolaten testades också med Omnilog PM-‐paneler med olika antibiotika av intresse. Ett antal isolat valdes även för molekylära tester med PCR där man sökte efter de plasmidburna resistensgenerna CTX-‐M1, CTX-‐M2 och SHV. Data från tester på den totala mikrofloran var svår att tolka; för att utveckla metoden till ett tillförlitligt verktyg krävs vidare optimering. För alla test med PM-‐plattor (total microbiota och enskilda isolat) visades största skillnader mellan olika antibiotika i brunnar med starkast koncentration. Totalt uppvisade 175 isolat förmåga att växa på ESBL-‐agar. E-‐tester bekräftade resistens hos isolat mot β-‐laktam-‐antibiotika. Många av dessa var kliniskt resistenta. Inga plasmider överfördes under exogen isolering. Under molekylära tester har några träffar för CTX-‐M1 genen hos de utvalda isolaten, men inte för de andra generna. Isolaten som analyserades med Omnilog PM-‐plattor visade resistens mot de flesta testade substanserna. Isolaten visade störst resistens mot penicillin G, ampicillin, amikacin, kanamycin och trimetoprim. I studien konstaterades att resistens bland bakterier var vanligt i bevattningsvattnet och att bakterier resistenta mot β-‐laktam-‐antibiotika var mycket vanligt. Vidare studier behövs för att bedöma frekvens-‐och förekomsten av horisontell genöverföring (HGF) i denna miljö. Även angående utredning av orsak till uppkomsten av resistens krävs mer undersökningar. Data som erhållits skulle kunna visar att resistens finns mot substanser som är vanliga inom veterinärmedicin (t ex penicillin G). Dock är dessa substanser även vanliga inom humanmedicin vilket gör att det inte går att dra någon slutsats angående resistensens ursprung utifrån data i denna studie. Spridning av bakterier med ESBL-‐resistens har även visats ske via människor som exponeras-‐ och blir bärare av resistenta bakterier via exempelvis resor. Studier visar också att resistenta bakterier sprids via transport-‐ och handel med varor och djur. SLU/Plant Breeding and Biotechnology (until 121231) 2013 H2 eng swe https://stud.epsilon.slu.se/5279/ |
| spellingShingle | Animal husbandry Enterobacteriaceae ESBL Etest exogenous isolation irrigation water hygiene standards in water Water quality Grudén, Maria Antibiotic resistance associated with bacteria in irrigation water : a case study of irrigation ponds in Southern Sweden |
| title | Antibiotic resistance associated with bacteria
in irrigation water : a case study of irrigation ponds in Southern Sweden |
| title_full | Antibiotic resistance associated with bacteria
in irrigation water : a case study of irrigation ponds in Southern Sweden |
| title_fullStr | Antibiotic resistance associated with bacteria
in irrigation water : a case study of irrigation ponds in Southern Sweden |
| title_full_unstemmed | Antibiotic resistance associated with bacteria
in irrigation water : a case study of irrigation ponds in Southern Sweden |
| title_short | Antibiotic resistance associated with bacteria
in irrigation water : a case study of irrigation ponds in Southern Sweden |
| title_sort | antibiotic resistance associated with bacteria
in irrigation water : a case study of irrigation ponds in southern sweden |
| topic | Animal husbandry Enterobacteriaceae ESBL Etest exogenous isolation irrigation water hygiene standards in water Water quality |