Antibiotic resistance patterns in Escherichia coli from gulls in nine European countries.

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by  Stedt, Johan

(cid:2)THE GROWING GLOBAL PROBLEM OF ANTIBIOTIC RESISTANCE
Antibiotic resistance patterns in
Escherichia coli from gulls in nine
European countries
Johan Stedt, PhD Student1, Jonas Bonnedahl, MD, PhD1,2,
Jorge Hernandez, PhD Student1, Barry J. McMahon, PhD3,
Badrul Hasan, PhD4, Bjo¨rn Olsen, Professor4,
Mirva Drobni, PhD4 and Jonas Waldenstro¨m, PhD1*
1Centrefor Ecology andEvolution inMicrobial ModelSystems, Linnaeus University, Kalmar,Sweden;
2Department ofInfectious Diseases, Kalmar County Hospital,Kalmar,Sweden; 3Section ofClinical
Microbiology andInfectious Diseases, Department ofMedical Sciences,Uppsala University, Uppsala,
Sweden; 4UCDSchool of Agriculture&Food Science,University CollegeDublin, Belfield, Dublin, Ireland
Background:Theprevalenceofantibioticresistantfaecalindicatorbacteriafromhumansandfoodproduction
animalshasincreasedoverthelastdecades.InEurope,resistancelevelsinEscherichiacolifromthesesources
show a south-to-north gradient, with more widespread resistance in the Mediterranean region compared
tonorthernEurope.Recentstudiesshowthatresistancelevelscanbehighalsoinwildlife,butitisunknownto
whatextentresistancelevelsinnatureconformtothepatternsobservedinhuman-associatedbacteria.
Methods:Totestthis,wecollected3,158faecalsamplesfrombreedinggulls(Larussp.)fromnineEuropean
countries and tested 2,210 randomly isolated E. coli for resistanceagainst 10antibiotics commonly used in
human andveterinary medicine.
Results: Overall, 31.5% of the gull E. coli isolates were resistant to ]1 antibiotic, but with considerable
variation between countries: highest levels of isolates resistant to ]1 antibiotic were observed in Spain
(61.2%)andlowestlevelsinDenmark(8.3%).Foreachtestedantibiotic,theIberiancountrieswereeitherthe
countries with the highest levels or in the upper range in between-country comparisons, while northern
countries generally had a lower proportion of resistant E. coli isolates, thereby resembling the gradient of
resistance seen inhuman and foodanimal sources.
Conclusion: We propose that gulls may serve as a sentinel of environmental levels of antibiotic resistant
E. colito complement studies of human-associated microbiota.
Keywords: EscherichiaColi;wildlife;birds;gulls;Antibioticresistance;antibiotics
Received:28May2013;Revised:29November2013;Accepted:29November2013;Published:10January2014
Antibiotic resistant bacteria are a global problem coli collections from human and veterinary sources cor-
for both human and animal health (1(cid:2)3). For relates with the antibiotic consumption within sampled
instance, in Europe, the European Centre for regions(7,8).Inreactiontothissituation,theEuropean
Disease Prevention and Control (ECDC) estimated that Union has evoked measures to reduce unnecessary
the death toll from nosocomial infections with resistant usage of antibiotics in the member countries (9, 10).
bacteria exceeds 25,000 patients per year (4). The levels Forexample,since2006antibioticsarebannedasgrowth
of antibiotic resistance have also increased in indicator promoters in food production animals (EC regulation
bacteria from commercially reared swine, poultry and 1831/2003).
cattlepopulationsinEurope(5),andtheresistancelevels The gram-negative bacterium E. coli is the most
inindicatorbacteriafromthesefoodproductionanimals frequently used indicator bacterium for addressing anti-
correlate with resistance levels in human samples at the biotic resistance dissemination in different environments
nationallevel,indicatingregulartransmissionofbacteria and host species, as it is a frequent carrier of different
andresistancegenes(6).Furthermore,therearestrongin- antibiotic resistance genes and a prominent constituent
dications that the occurrence of resistance in Escherichia of human and most mammal and bird gastrointestinal
InfectionEcologyandEpidemiology2014.#2014JohanStedtetal.ThisisanOpenAccessarticledistributedunderthetermsoftheCreativeCommons 1
Attribution3.0UnportedLicense(http://creativecommons.org/licenses/by-nc/3.0/),permittingallnon-commercialuse,distribution,andreproductioninany
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Citation:InfectionEcologyandEpidemiology2014,4:21565-http://dx.doi.org/10.3402/iee.v4.21565
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JohanStedtetal.
microbiota (11(cid:2)13). Based on data from human E. coli, recent years (22(cid:2)25, 28(cid:2)33). However, most of these
a south-to-north gradient in resistance levels has been studies have been restricted either in the geographical
observed in Europe, with greatest levels of isolates with coverage of the sampling areaor in the time span of the
resistance in the Mediterranean countries and lowest investigation, making large-scale inferences and compar-
levels in the Nordic countries (14, 15). isons between studies difficult. In the present study,
Human and food animal bacteriawith resistant geno- we sampled gulls in nine different European countries
typesarelikelyfrequentlydisseminatedinto theenviron- simultaneously during the late part of the breeding
ment, via sewage water, for example (16, 17). season, allowing forcomparisons of antibiotic resistance
In recent years, several reports of antibiotic resistant levelsinE.colibetweencountrieswithdifferingantibiotic
bacteria from both environmental and wildlife sources resistance levels. As focal species, weused closely related
havebeenpublished(12,13,18),andE.coliwithresistance gull species, the Herring gull (Larus argentatus) and the
genes have been isolated from several animal species, Lesser Black-backed gull (Larus fuscus) that occur in
including wild boars (19), brown rats (20) and Gilthead NorthernandWesternEurope,andtheirsisterspeciesthe
sea bream (21). Furthermore, similar E. coli sequence Yellow-legged gull (Larus michahellis) that occurs in
types(STs)orclonalgroupsareoftenisolatedfromgulls, SouthernandSouthwesternEurope(34).Thesegulls are
humansanddomesticatedanimals(22(cid:2)25).Generally,in widespread,havesimilarfeedingecologyandarecommon
smallerbirds,suchaspasserines,resistantE.coliarerare inmanmadeenvironments,especiallyintowns,harbours
orabsent,andstudiesmainlyexaminingspecificresistance and areas with trash. Our aim was to investigate the
markers such as extended-spectrum beta lactamases spatialvariationingullE.coliresistancelevelsinacross-
(ESBL)havebeenpublished(26,27).Largerbirdspecies, sectionalsurveyacrossEuropetoevaluatethedissemina-
especially those that live in close proximity to humans, tion of resistant E. coli in the environment. If the
such as crows, other corvids and gulls seem to be more antibiotic resistance levels in gulls are mainly reflecting
frequentcarriersofE.coliresistanttoantibioticscommon thesituationintheareawheretheybred,wehypothesize
inhumanandlivestock(13).Gullsandcorvidsutilizefood thatantibioticresistancelevelsingullsshouldconformto
resources made available by humans, and often feed patternsobservedinothersources.
inurban areas, at garbagedumps, at sewage plants or in
agricultural areas rich in livestock. As a consequence, Material and methods
resistant E. coli can be locally high in some wild bird
populationsassociatedwithhumans(23,24). Study species, sampling sites and methodology
Thepresenceofresistantbacteriaingullshasprompted Sampling was conducted from mid-June to early July
their use as an indicator of environmental antibiotic 2009, in the late part of the gull-breeding period,
resistance occurrence, and there have been several pub- where three sampling teams visited eight different
lications presenting data from various gull species in European countries, sampling Herring gulls and Lesser
Table1. Summaryofsampling sites, sampled gull species, numberofcollected samples, isolatedE.coliincluded in thestudyanda
descriptionofthesitessampled.
Country Location Species Samplesn(IsolatedE.coli) Description
Denmark Bla˚vand Larusfuscus(60%), 158(96) CoastlineclosetoRoskilde
Larusargentatus(40%)
England Bristol Larusargentatus 133(68) CentralBristol
Ireland Howth Larusargentatus 266(239) SuburbofDublincontainingfishingharbour
Latvia Daugava Larusargentatus 323(224) ClosetocentralRiga
Latvia Kaltene Larusargentatus 101(60) Countryside60kmfromRiga
Netherlands Borsele Larusargentatus 280(229) Industryharbour
Netherlands Rotterdam Larusfuscus(90%), 280(228) Industryharbour
Larusargentatus(10%)
Poland Wladyslawowo Larusargentatus 280(235) CoastlineclosetoGdansk
Portugal Lagos Larusmichaellis 314(176) FishmarketinLagos
Portugal Portimao Larusmichaellis 111(76) CoastlineclosetoPortimao
Spain Emporda Larusmichaellis 199(84) CitydumpoutsideEmporda
Spain Mazarron Larusmichaellis 321(200) BreedingcolonyclosetoMazarron
Spain Malaga Larusmichaellis 175(120) HarbourinMalaga
Sweden Hudiksvall Larusargentatus 217(175) ArchipelagooutsidethecityofHudiksvall
2
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AntibioticresistancepatternsinEscherichiacoli
Black-backed gulls in northern Europe, and Yellow- tion, 2,210 E. coli isolates remained, representing an
legged gulls in southern Europe (Table 1). Additionally, isolationfrequencyof42(cid:2)90%ofthetotalsampleineach
inIrelandoneoftheauthors(BMcM)carriedoutsampl- country(Table1).
ing from June to August. In collaboration with local The susceptibility of each isolate was tested with disc
ornithologistsineachcountry,thefieldpersonnelvisited diffusionagainstapanelof10antibioticagentsusedinE.
localitieswithlargenumbersofbreedinggulls.Fromeach coli infections in human and veterinary medicine: ampi-
site, 50(cid:2)450 samples were collected. Sampling sites were cillin 10 mg/disc, cefadroxil 30 mg/disc, chloramphenicol
chosen to be as similar as possible between countries, 30 mg/disc, nalidixic acid 30 mg/disc, nitrofurantoin 100
taking human activity and gull feeding habitats into mg/disc, mecillinam 10 mg/disc, tetracycline 30 mg/disc,
consideration. tigecycline 15 mg/disc, streptomycin 10 mg/disc and
For sampling, a sterile cotton wool swab was swirled trimethoprim/sulfamethoxazole25mg/disc(allantibiotics
in freshly deposited faecal material from the ground from Oxoid Ltd, Cambridge, UK). Susceptibility testing
wheregullshadrecentlybeenroosting.Toavoidmultiple was conducted using clinical breakpoints on Mueller-
sampling from the same bird, samples were collected Hinton agar according to EUCAST recommendations,
from dispersed flocks and always in less numbers than including the E. coli ATCC 25922 as a reference strain.
therewereindividualspresentateachsite.Allswabswere Currently, tigecycline and streptomycin lack defined
inoculated into tubes containing a bacterial freeze med- EUCAST breakpoints for susceptibility testing; there-
ium(Luriabroth; BD,Sparks, USA,phosphatebuffered fore, breakpoints for these antibiotics were defined
saline containing 0.45% Na-citrate, 0.1% MgSO , 1% according to the NRI method (35).
4
(NH ) SO , and 4.4% glycerol) and immediately frozen
4 2 4
andstoredinliquidnitrogeninthefield,andlaterinthe Results
laboratory at (cid:3)808C. All samples were collected from
adultandsub-adultbirdsinoraroundbreedingcolonies. Antibiotic susceptibility of randomly selected E. coli
In total, 31.5% of the 2,210 tested E. coli isolates were
Isolation of E. coli and antibiotic resistance resistant to ]1 antibiotic, and 19% of the isolates
testing displayed resistance to ]2 antibiotics (Table 2). In
Intotal,3,158faecalsampleswithbacterialgrowth(101(cid:2) general, tetracycline resistance was the most common
323samples/location)wereavailableforanalysis(Table1). phenotype (19.0%) of E. coli isolates, followed by resis-
EachsamplewasplatedonanUriselect4plate(Bio-Rad tance to ampicillin (18.1%), while none of the isolates
Laboratories Ltd, Hemel Hempstead, UK) for isolation were resistant to tigecycline, and very few to mecillinam
of putative E. coli and one randomly chosen colony per (0.7%)(Fig.1,Table3).Thelevelofantibioticresistance,
samplewas purified and further isolated. E. coli identity measuredasthenumberofisolateswithresistanceto ]1
was confirmed using conventional biochemical testing antibiotic, varied between countries (x2(cid:4)626.2 df(cid:4)8,
(urease-negative, H S-negative, indole-positive and b- pB0.001),withhighestobservedlevelsinSpain(61.2%),
2
galactosidase-positive). After isolation and characteriza- England (44.1%) and Latvia (38.7%). Lowest resistance
Table2. TheproportionofE.coliisolateswithresistancetooneormoreantibioticsoneachsamplingsite.
0 ]1 ]2 ]3 ]4 ]5 ]6 ]7 8
Denmark 91.7 8.3 5.2 0.0 0.0 0.0 0.0 0.0 0.0
England 55.9 44.1 25 13.2 7.4 1.5 2.9 1.5 0.0
Ireland 88.4 11.6 1.9 0.4 0.0 0.0 0.0 0.0 0.0
Latvia,Daugava 58.0 42.0 31.7 26.3 18.7 12.5 40.0 0.4 0.0
Latvia,Kaltene 96.7 3.3 0.0 0.0 0.0 0.0 0.0 0.0 0.0
Netherlands,Borsele 75.5 24.5 10.9 7.0 3.5 1.7 0.0 0.0 0.0
Netherlands,Rotterdam 83.3 16.7 9.2 7.4 5.2 2.6 2.2 0.4 0.0
Poland 78.7 21.3 4.7 2.6 1.7 0.9 0.0 0.0 0.0
Portugal,Lagos 76.4 23.6 18.3 11.0 9.4 7.3 2.1 0.0 0.0
Portugal,Portimao 69.1 30.9 24.7 16.0 9.9 3.7 1.2 0.0 0.0
Spain,Emporda 39.3 60.7 40.5 28.6 20.2 10.7 2.4 1.2 0.0
Spain,Mazzaron 32.9 67.1 57.3 45.3 31.5 16.9 5.8 0.9 0.0
Spain,Malaga 50.0 50.0 42.5 35.0 27.5 15.0 7.5 0.8 0.0
Sweden 82.3 17.7 9.7 6.9 4.0 3.4 1.1 0.0 0.0
3
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Fig.1. GeographicalvariationingullE.coliresistancelevelsinninedifferentEuropeancountries.Dotsrepresentsamplingsitesineach
country,andthebarplotsshowaverageresistancelevels(%)toeachoftheanalysedantibiotics.Theantibiotics,fromlefttotheright:
nalidixicacid(NA),streptomycin(S),tetracycline(TE),ampicillin(AMP),chloramphenicol(C),cefadroxil(CFR),tigecycline(TGC),
nitrofurantoin(N),mecillinam(MEL)andtrimethoprim/sulfamethoxazole(SXT).ThebarplotfromSpainisenlargedtoindicatethe
scaleused.
levelswere seen in Denmark (8.3%) and Ireland (11.6%) there were no evident differences in resistance levels
(Table 3). depending on sample location.
Similarly, there were also large differences in resis-
tance levels between countries when each antibiotic was Discussion
analysedseparately(Table3).Greatestresistancelevelsat
the country level were to ampicillin (42.7%) and tetra- Spatial variation in gull E. coli resistance levels
cycline (44.1%) in Spain, but sixof the tested antibiotics across Europe
hadresistancelevels(cid:2)15%inatleastonecountry(Table3). This study demonstrates that the spread of antibiotic
Spain also had the highest proportion (6.5%) of isolates resistant E. coli is not confined to clinical and food pro-
withresistanceto ]6antibiotics,incontrasttoDenmark, duction environments, as high rates of E. coli resistance
PolandandIrelandgullsamplesthathadnoisolatewith to ]1 antibiotic were frequently found in the faecal
resistance to ]6 antibiotics (Table 2). In total, 59 dif- microbiotaofgulls.Insixofthecountries, (cid:2)20%ofthe
ferentphenotypeswithresistanceto ]3antibioticswere E.colifromthegullswasresistantto ]1antibiotic,and
found. At the local scale, the most striking variation in in Spain more than half of the isolates had resistant
resistance levels was found in Latvia where 224 E. coli phenotypes. The diversityof multiresistant (]3 antibio-
isolated from agull-breeding colony situated in the river tics)phenotypeswithinthetestedE.coliwassubstantial,
Daugava, downstream of central Riga, had higher resis- with 59 different resistance combinations detected
tance levels than the corresponding 80 E. coli isolates (Table 4). The large diversity in resistance phenotypes
from gulls breeding in the countryside 100 km outside both within and between sites indicates that more than
Riga (Fig. 2). In other countries with more than one oneclonepercountrywasinvolved.Highratesofgeneral
samplesite,suchasSpain,PortugalandTheNetherlands, antibioticresistance ingullE.colihavebeenobservedin
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AntibioticresistancepatternsinEscherichiacoli
e
ol previous studies. For example, observed resistance was
z
xa 47.1% to ]1 antibiotic in E. coli isolated from Yellow-
o
h legged gulls in southern France (24). In addition, E. coli
et
m from Black-headed gulls Larus ridibundus sampled in
ulfa 2.1 0.3 0.8 7.4 0.0 5.7 5.7 0.9 8.4 2.3 5.5 9.8 6.6 4.6 Sweden and the Czech Republic had resistance levels of
s 1 1 1 1 2 1
m/ 13%and29%,respectively(25,30).However,ourstudyis
pri the first that investigates the distribution of antibiotic
o
h
et resistancecarriageacrossEuropeusingasingletaxaasa
m
Tri sentinel system.
Ingeneral,ourresultsconformedtothesouth-to-north
m
a gradient of resistance levels seen in E. coli samples from
cillin 0.0 1.5 0.4 0.4 0.0 0.4 0.0 0.0 0.0 2.5 1.2 0.4 0.8 0.0 humansandfoodproductionanimalsacrossEurope(15),
e
M but with considerable variation between countries in
different regions. For each tested antibiotic, the Iberian
n
ntoi countrieswereintheupperrangeinlevelsofE.coliwith
ura 0.0 0.0 0.4 4.0 0.0 0.0 0.0 0.0 1.0 0.0 0.0 1.3 0.0 0.0 resistance in between-country comparisons, while coun-
Nitrof tarniedsDinenthmeanrko)rthhaedrnassammaplllienrgparorepaor(tfioorninosftaisnocleatIersewlainthd
ne resistance. Additionally, antibiotics with overall low
gsite. gecycli 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0 3.3 0.0 rFeosrisetaxnacmepllee,v1el.s2%shoofwtehdeissiomlailtaerdEge.ocgorlaipwhaiscarlespisattatnertntso.
plin Ti nitrofurantoin in Spain and Portugal, while it was not
m xil found at all in Sweden, Denmark, England, Poland and
persa efadro 0.0 1.5 0.8 1.8 0.0 0.4 0.9 0.0 3.1 0.0 3.6 11.1 10.8 2.3 TnohteaNbseothluetrela,nadnsd(sToambleec3o).unHtroiwese,vseur,chthaesgSrwadeideennt awnads
d C
parate nicol Econmglpaanrde,dhatdohoigthheerrrecsoiustnatnrcieesleoveflstihnethseamguelllsaatmitupdlees,
ntibioticse hloramphe 0.0 2.9 0.4 10.3 0.0 0.4 2.6 0.4 3.7 4.9 10.7 20.4 16.7 2.9 stiuogTng,heeostrliansraggmeesiptthldienirfgfreeerfgefineoccntesa.almoronlogcsailtevsawriiatthioinnaincoduisnstermyiwnaas-
a C seeninLatvia,wherethegullpopulationsampledcloseto
d
e n the capital Riga had higher resistance levels than E. coli
thetest Ampicilli 6.2 33.8 1.9 29.9 1.7 13.5 8.8 6.0 15.7 17.3 39.3 44.9 42.5 13.7 fTrhoims stuhgegpeostpsutlhataitonthseadmepglreedeoonf athnethcrooupnotgreynsiidcein(Ffliuge.n2c)e.
of canalsovaryonasmallspatialscale.Italsohighlightsthe
h ne
ncetoeac Tetracykli 1.0 27.9 2.7 29.0 0.0 15.3 12.3 6.0 18.3 25.9 34.5 52 37.5 10.3 uulassreegfeutrhlnessepesasthioaafblisutcataitllsiezsai.nsIgnsernSivtpeianriencls,atPocohfrmatunegtniatblsioaatnnidcdrTsephsieesctNiaensecthteheaartt-
a lands, where samples were also collected from different
esist ycin locations,wedidnotdetectanysignificant differencesin
r m 1 3 8 4 7 6 9 4 4 8 4 7 9
with epto 3. 10. 0. 21. 1. 9. 7. 3. 9. 14. 27. 28 21. 6. rtheseissetansicteeslebveeilnsgbemtwoereenssimamilaprlinignsfieteesd,inpgerhhaapbsitadtus,e otor
oli Str humandensity.Weaimedtostandardisesamplingsitesin
c
E. d allcountriesasfaraspossible.Withtheexceptionofone
ci
atedgull alidixica 0.0 5.9 0.8 21.4 0.0 2.2 5.7 3.4 12.0 8.6 32.1 36.9 30.8 2.9 switeeLreoincookLlilnaetgcvteiaadtacsnlpodes,ceitftoiocslraoermsgiesetraecnxictteieenstp.rSowfieledse,nt,htehemsoasmtpfrlees-
ol N
s quently recovered phenotypes were resistant to tetracy-
i m
of a cline or ampicillin. These results are in concordance
Table3.Percentage Denmark England Ireland Latvia,Daugava Latvia,Kaltene Netherlands,Borsele Netherlands,Rotterd Poland Portugal,Lagos Portugal,Portimao Spain,Emporda Spain,Mazzaron Spain,Malaga Sweden wtiolMnihncietceabhucmfoiraltorlchieattnhnnvatecdehmretaneostratfriutlneihoddsaetiiirhssexyteseiacranaonrancnedcetsiigibdhsuiitusoralmletnssisccait(ssein2ltla4hpmn,ahrcvee2aeedn5r,eio,bcwtie3yniae0npesn)emfsacco,lonorisnmdrudeiccmemhascoualaanrdyspilesyrrmsoie.sulfeTailsncetheiegcldest-.
5
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Nalidixic acid
Streptomycin
30
Tetracycline
Ampicillin
Chloramphenicol
Cefadroxil
Tigecyline
Nitrofurantoin
20 Mecillinam
%)
Sulfametoxazole
nt (
e
c
r
e
P
10
0
Riga Country side
Fig.2. DifferencesinantibioticresistancelevelsbetweentwosamplingsitesinLatvia.BarsshowsthepercentageofisolatedE.coliwith
resistancetoeachtestedantibiotic.
Table4. Allcombinationsofrecognisedphenotypeswithmultiresistance(]3antibiotics).Illustratingthediversityofmultiresistant
phenotypesamongthetestedE.colifromtheninesampledcountriesacrossEurope.
Poland Sweden Latvia Ireland England Netherlands Spain Portugal Denmark Total
NA-S-TE 0 0 2 0 0 0 2 0 0 4
NA-S-AMP 0 0 3 0 0 0 0 1 0 4
NA-S-C 0 0 0 0 0 0 1 0 0 1
NA-TE-AMP 0 1 5 0 0 3 12 1 0 22
NA-TE-C 0 0 0 0 0 0 0 1 0 1
NA-TE-F 0 0 0 0 0 0 1 0 0 1
NA-TE-SXT 0 0 0 0 0 0 1 0 0 1
NA-AMP-CFR 0 0 0 0 0 0 3 0 0 3
NA-AMP-SXT 0 0 2 0 0 0 1 0 0 3
TE-AMP-C 0 0 1 0 0 0 0 0 0 1
TE-AMP-CFR 0 0 0 0 0 0 3 1 0 4
TE-AMP-MEL 0 0 0 0 1 1 0 1 0 3
TE-AMP-SXT 0 1 0 0 2 1 2 0 0 6
TE-C-SXT 0 1 0 0 0 0 1 3 0 5
S-AMP-SXT 1 0 3 0 1 0 1 0 0 6
S-TE-AMP 1 1 0 0 0 7 8 0 0 17
S-TE-C 0 1 0 0 0 0 0 0 0 1
S-TE-SXT 0 0 1 0 0 1 8 0 0 10
NA-S-TE-AMP 3 0 3 0 0 0 8 1 0 15
NA-S-TE-F 0 0 0 0 0 0 1 0 0 1
NA-S-TE-SXT 0 0 0 0 0 0 2 0 0 2
NA-S-AMP-SXT 0 0 1 0 0 1 4 0 0 6
NA-TE-C-SXT 0 0 1 0 0 0 0 0 0 1
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AntibioticresistancepatternsinEscherichiacoli
Table4(Continued)
Poland Sweden Latvia Ireland England Netherlands Spain Portugal Denmark Total
NA-TE-AMP-SXT 0 0 0 0 0 1 7 0 0 8
NA-TE-AMP-C 0 0 0 0 0 0 10 2 0 12
NA-TE-AMP-CFR 0 0 0 0 0 0 2 0 0 2
NA-TE-AMP-MEL 0 0 0 0 0 0 1 0 0 1
NA-AMP-C-CFR 0 0 0 0 0 0 2 0 0 2
NA-AMP-C-F 0 0 1 0 0 0 0 0 0 1
NA-AMP-C-SXT 0 0 0 0 0 0 2 0 0 2
S-TE-AMP-CFR 0 0 1 0 0 0 0 0 0 1
S-TE-AMP-C 0 0 0 0 1 0 0 1 0 2
S-TE-AMP-SXT 0 1 7 0 2 8 12 1 0 31
S-TE-C-SXT 0 0 0 0 0 0 2 2 0 4
S-AMP-MEL-SXT 0 0 0 0 0 0 0 1 0 1
TE-AMP-C-CFR 0 0 0 0 0 0 1 0 0 1
TE-AMP-C-SXT 0 0 0 0 0 0 3 1 0 4
NA-S-TE-AMP-C 0 0 1 0 0 0 4 0 0 5
NA-S-TE-AMP-CFR 0 3 0 0 0 0 1 0 0 4
NA-S-TE-AMP-F 0 0 0 0 0 0 1 0 0 1
NA-S-TE-AMP-SXT 1 1 5 0 0 3 12 10 0 32
NA-S-AMP-F-SXT 0 0 0 0 0 0 1 0 0 1
NA-TE-AMP-CFR-SXT 0 0 1 0 0 0 2 0 0 3
NA-TE-AMP-C-CFR 0 0 0 0 0 0 10 1 0 11
NA-TE-AMP-C-F 0 0 5 0 0 0 0 0 0 5
NA-TE-AMP-C-SXT 0 0 5 0 0 1 1 0 0 7
NA-TE-C-F-SXT 0 0 1 0 0 0 0 0 0 1
S-TE-AMP-C-SXT 0 0 1 0 0 0 6 0 0 7
S-TE-AMP-CFR-SXT 0 0 0 0 0 1 1 0 0 2
S-TE-AMP-MEL-SXT 0 0 0 0 0 0 1 0 0 1
S-TE-AMP-F-SXT 0 0 0 0 0 0 0 1 0 1
TE-AMP-C-CFR-SXT 0 0 0 0 0 0 1 0 0 1
NA-S-TE-AMP-C-SXT 0 2 7 0 1 4 13 3 0 30
NA-S-TE-AMP-F-SXT 0 0 1 0 0 0 0 0 0 1
NA-S-TE-AMP-CFR-SXR 0 0 0 0 1 0 2 2 0 5
NA-S-TE-AMP-C-F-SXT 0 0 1 0 0 0 0 0 0 1
NA-S-TE-AMP-C-CFR 0 0 0 0 0 0 3 0 0 3
NA-S-AMP-C-CFR-SXT 0 0 0 0 0 0 1 0 0 1
NA-S-TE-AMP-C-CFR-SXT 0 0 0 0 0 1 4 0 0 5
NA, nalidixic acid; S, streptomycin; TE, tetracycline; AMP, ampicillin; C, chloramphenicol; CFR, cefadroxil; TGC, tigecycline; N, nitro-
furantoin;MEL,mecillinamandSXT,trimethoprim/sulfamethoxazole.
(36, 37), but was detected in gull E. coli in six countries, negative pathogens; consequently, the resistance levels
albeit at low levels. Nalidixic acid resistance has been to tigecycline are still low in human populations (39).
previously found in E. coli from wildlife, for instance, in
Yellow-legged gulls (25) and Mallards (31), but the very Origin of resistance?
highratesofresistancetothisantibioticinSpanishgullE. Therearecomparativelyfewstudiesthatpresentdataon
coliwereremarkableandcomparable,orevenhigher,than general resistance levels from E. coli isolates at larger
thelevelsseeninhumanclinicalisolatesinSpain(36,38). spatial scales in Europe. EFSA publishes ayearly report
Tigecycline was the only tested antibiotic that we did on resistance data for bacteria from food production
notfindanyresistanceto.Itisarelativelynewantibiotic, animalsthatcoverslargepartsofEurope.Arecentstudy
used for skin-structure infections and complicated intra- compared resistance in human E. coli from the EARSS
abdominal infections, and has abroad spectrum activity scheme with EFSA E. coli resistance data from food
against multidrug-resistant Gram-positive and Gram- production animals, which showed that resistance levels
7
Citation:InfectionEcologyandEpidemiology2014,4:21565-http://dx.doi.org/10.3402/iee.v4.21565
(pagenumbernotforcitationpurpose)
JohanStedtetal.
infoodproductionanimals(especiallypoultryandswine) thereforehaveoriginatedfromdifferentlocations,obscur-
E. coli and human E. coli correlated at the country level ing the link between gulls carrying antibiotic resistant
(6). A key issue for the future is to increase monitoring bacteria and a particular region, and sampling should
schemes with standardised methodologies for measuring therefore be performed during breeding times prior to
and interpreting antibiotic resistance in bacteria from migration.Animportantquestiontoaskisforhowlong
humans, food production animals and the environment. do gulls shed resistant bacteria, as this figure would
Inthisregard,EFSAalreadyhasanongoingharmonised influence the potential of birds introducing resistant
monitoring system (40), while suitablereferencematerial bacteriatonewareasfarfromtheorigin.ForSalmonella,
of single antibiotics from human indicator bacteria is thesheddingtimeinHerringgullsisshort,onlyafewdays
lacking. Without such baseline data, our understanding (42), while some empirical data suggest that Salmonella
of antibiotic resistance dissemination could beobscured. can survive in the soil of gull nesting colonies between
From available data, it has been shown that Sweden breeding seasons (43). Furthermore, chickens can carry
andTheNetherlandshavesimilarlowantibioticusagein resistant E. coli for several weeks (44), and it has been
thehumanhealthcaresector(14),andalsoacomparable suggested that gulls and other birds bring antibiotic
low resistance level among E. coli from humans (15). resistant bacteria to very remote areas in the Arctic,
However, antibiotic usage in food production is much indicatinglongersheddingtime(22,45).
higher in The Netherlands, and concomitant high resis-
tance levels are seen in E. coli from Dutch food animal
Acknowledgements
sources (5, 41). It is interesting to note that this pattern
Wewouldliketoextendourthankstothosewhomadeitpossible
was also seen in the gull samples, where resistance levels
tosamplelargepartsofEurope:A.Eriksson,G.Norevik,A.Steen,
tosingleantibioticswerehigherinTheNetherlandsthan
M. Stervander, P. O¨sterman, M. Axbrink, R. J. Buijs, P. Feliu,
in Sweden for five antibiotics, including tetracycline and
P. Andersen Harild, S. Garcia, P. Rock, D. Rodrigues, A. Cama
streptomycin, while Sweden only had higher levels to TorellandS.Wates.
cefadroxil and chloramphenicol. These differences indi-
cate that food production could be important for the Conflict of interest and funding
dissemination of resistant bacteria to the environment.
This work was supported by grants from the Swedish
However, at the same time, The Netherlands has a far
Research Council FORMAS (2008-326), The Karin
greater population density than Sweden, thereby poten-
Korsner’s Foundation and The Olle Engkvist Byggma¨s-
tially increasing the likelihood of transmission from
tare Foundation.
human or food-producing animals to wildlife, which
could further disseminate resistance. The mechanisms of
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