open access faecal occurrence and emissions of live- stock

Berliner und Münchener Tierärztliche Wochenschrift 126, Heft 3/4 (2013), Seiten 117–180 117

U.S. Copyright Clearance Center Code Statement: 0007-9366/2013/12603-117 $ 17.00/0

Open Access

Berl Münch Tierärztl Wochenschr 126, 117–180 (2013)DOI 10.2316/0007-9366-126-117

© 2013 Schlütersche Verlagsgesellschaft mbH & Co. KGISSN 0007-9366

Korrespondenzadresse: [email protected]

Eingegangen: 02.06.2012 Angenommen: 22.08.2012

Online first: 08.03.2013http://vetline.de/zeitschriften/bmtw/open_access.htm

Institute for Animal Hygiene and Environmental Health, Free University Berlin, Germany1

Institute for Animal Hygiene, Animal Welfare and Farm Animal Behaviour, University of Veterinary Medicine Hannover, Foundation, Germany2

Faecal occurrence and emissions of live-stock-associated methicillin-resistant Staphylococcus aureus (laMRSA) and ESBL/AmpC-producing E. coli from animal farms in Germany

Vorkommen livestock-assoziierter Methicillin-resistenter Staphylococcus aureus (laMRSA) und ESBL/AmpC-produ-zierender E. coli im Kot und deren Emission aus Nutztier- ställen in Deutschland

Anika Friese1, Jochen Schulz2, Henriette Laube1, Christina von Salviati1, Joerg Hartung2, Uwe Roesler2

Summary The occurrence of laMRSA (livestock-associated methicillin-resistant Staphylococcus aureus) and extended-spectrum β-lactamase (ESBL) and/or plasmid-mediated AmpC β-lactamase-producing (AmpC) Enterobacteriaceae in healthy livestock herds is known for some time. The spread of these bacteria in the environment is discussed critically. The object of this study was to determine the presence of these microorganisms in faeces of livestock as well as the discussion about a potential faecal emission. Therefore, faeces samples from 31 different MRSA positive livestock holdings were tested for MRSA. Furthermore, faeces sam-ples from 70 farms with an unknown status regarding ESBL/AmpC-producing E. coli were screened for those resistant bacteria. LaMRSA was detected in samples of turkey (2/7, 40%) and broiler fattening farms (1/4, 27%) as well as in pig farms with higher detection frequencies in fattening farms (11/17, 13.3%) than in breed-ing farms (4/12, 33.3%). ESBL/AmpC-producing E. coli was found in all investi-gated eight broiler farms (100%), in nine out of 16 (76.3%) breeding pig as well as in six out of 10 (60%) dairy cattle herds and in seven of 16 (43.8%) fattening pig holdings. This presents the first detection of ESBL/AmpC-producing E. coli origi-nating from healthy pigs, turkeys and broilers in Germany. In addition, samples of fertilized field surfaces were studied exemplarily for the presence of MRSA (n = 4) as well as ESBL/AmpC-producing E. coli (n = 2). Furthermore, slurry samples from four broiler and five pig farms were analysed for the latter. Both MRSA and ESBL/AmpC-producing E. coli could be detected on the field surfaces, the last also in slurry samples. Faecal emissions from animal husbandry seem to be one possible route for the spread of these resistant microorganisms in the environment.

Keywords: resistant microorganisms, MRSA, ESBL, livestock, environmental health, slurry

Zusammenfassung Das Vorkommen von laMRSA (livestock-assoziierte Methicillin-resistente Staphylococcus aureus) und ESBL (Extended Spectrum β-Lactamase) und/oder plasmid-vermittelten AmpC-β-Lactamase-produzierenden (AmpC) Enterobacteriaceae in gesunden Nutztierbeständen ist seit einiger Zeit bekannt. Eine mögliche Ausbreitung dieser Keime in die Stallumgebung wird zunehmend kritisch in der Öffentlichkeit betrachtet. Ziel dieser Studie ist die Erfassung des Vorkommens die-ser Mikroorganismen im Kot verschiedener Nutztiere sowie die Diskussion einer möglichen Verbreitung der resistenten Bakterien über fäkale Emissionen. Dazu wurden Sammelkotproben von 31 verschiedenen MRSA-positiven Nutztierbe-ständen auf das Vorkommen von MRSA untersucht. Weiterhin wurden Kotproben aus 70 Beständen mit unbekanntem Status bezüglich ESBL/AmpC-produzie-renden E. coli auf diese Keime untersucht. MRSA konnten sowohl in einigen Proben der Putenbestände (2/7, 40 %) und Broilermastbestände (1/4, 27 %) als

BUM_3_4_s175-180_12044_Friese_pr 175 26.02.13 10:12


auch in denen der Schweinebestände, dabei mit höheren Nachweisraten in Mast- (11/17, 13,3 %) als in Zuchtbetrieben (4/12, 33,3 %), gefunden werden. ESBL/AmpC-produzierende E. coli wurden in allen untersuchten Broilermastbeständen (8/8, 100 %), in 9/16 (76,3 %) der Schweinezucht- und 6/10 (60 %) der Milchrin-derbestände sowie in 1/16 (43,8 %) der Schweinemastbestände detektiert. Dies stellt gleichzeitig den ersten Nachweis von ESBL/AmpC-produzierenden E. coli bei gesunden Schweinen, Puten und Broilern in Deutschland dar. Zusätzlich wurden exemplarisch Proben der Bodenoberfläche von begüllten Feldern ausgewählter Bestände auf MRSA (n = 4) sowie ESBL/AmpC-produzierende E. coli (n = 2) unter-sucht. Weiterhin wurden Gülleproben von vier Geflügel- und fünf Schweinebe-ständen hinsichtlich ESBL/AmpC-produzierender E. coli analysiert. Sowohl MRSA als auch ESBL/AmpC-produzierende E. coli konnten auf den gedüngten Flächen nachgewiesen werden, Letztere auch in den Gülleproben. Das Ausbringen von Fäkalien zur Düngung könnte somit einen möglichen Weg des Austrags der resistenten Keime aus den Nutztierhaltungen darstellen.

Schlüsselwörter: Resistente Mikroorganismen, MRSA, ESBL, Nutztierhaltung, Umwelthygiene, Gülle

Introduction

Antibiotics are powerful therapeutic tools to treat bacte-rial diseases of humans and animals. However, constant application in human as well as in veterinary medicine accelerates the development of resistance in bacteria. Rising rates of resistant and multiresistant microorgan-isms in animal holdings are a steadily upcoming problem (Coque et al., 2008). Consequently, problems can arise in the treatment of infectious diseases caused by these resistant pathogens. Especially laMRSA (livestock-asso-ciated methicillin-resistant Staphylococcus aureus) and extended-spectrum N-lactamase (ESBL) and/or AmpC N-lactamase-producing (AmpC) Enterobacteriaceae in food producing animals are in the focus of public atten-tion and might be relevant for public health. In Germany, both laMRSA and ESBL/AmpC-producing Enterobacte-riaceae, occur in animal husbandry. LaMRSA could be found on German cattle farms with mastitis problems (Spohr et al., 2011), in healthy pigs (Meemken et al., 2008; Friese et al., 2012), turkey (Richter et al., 2012) and broilers (Friese et al., 2013). Also ESBL/AmpC-producing Enterobacteriaceae seem to be widespread among healthy livestock like pigs (Mesa et al., 2006 ), cattle (Liebana et al., 2006; Rodriguez et al., 2009) or poultry (Brinas et al., 2003; Mesa et al., 2006; Bortolaia et al., 2010; Overdevest et al., 2011) in Europe. Although published data for Ger-many are rare, ESBL/AmpC-producing E. coli could be detected in healthy cattle (Wieler et al. 2011a) and clini-cal samples of livestock (Ewers et al., 2012) as well as companion animals (Wieler et al., 2011b). The transfer of MRSA between livestock and humans has been reported earlier (Cuny et al., 2009; Graveland et al., 2011) and a transmission of ESBL-producing gram-negative bacteria between food-producing animals and humans via direct contact or meat is supposed (Smet et al., 2009). Another important topic is the spread of these pathogens into the environment, for example via the application of con-taminated faeces on fields as fertilizer. According to the federal statistical office in Germany liquid manure was applied to 5.9 Mio ha agricultural land and solid manure to 2.3 Mio ha in 2010. Therefore, faeces of pigs, cattle and

poultry are commonly used. This study presents data of two different, in parts ongoing, studies and discusses the capability of an emission of laMRSA and ESBL/AmpC-producing E. coli in the vicinity of animal farms in Germany.

Material and Methods

Animal farmsA total of 37 conventional farms (15 fattening pig, twelve breeding pig including weaner-to-grower-farms, five turkey, four broiler and one cattle farm) in Germany with a positive MRSA-status were examined for the presence of these bacteria in pooled faeces samples. The pig farms were distributed in the northern and eastern region of Germany, turkey farms in the southern and eastern part, the broiler farms were in the northwest as well as northeast and the cattle farm was in eastern Germany. A positive status of MRSA means that except the faecal sample at least one other sample of animals and/or environment inside the barn was tested positive for MRSA simultaneously. In addition to this, boot swabs taken from the surface of previously fertilized fields (within the last three months) around three selected pig farms and one turkey fattening farm were analysed. For pooled faeces samples around 250 g faeces from at least five different locations inside the barn were collected. For sampling the field ground surface a 50-metre distance was stepped with the boot swabs. These results are partly published in Friese et al. (2012) and Schulz et al. (2012). Furthermore, for the ESBL/AmpC study pooled faecal samples of 50 conventional farms with an unknown ESBL/AmpC status distributed regionally in the north-ern and eastern part of Germany (16 fattening pig, 16 breeding pig including weaner-to-grower-farms, eight broiler and ten dairy cattle farms), slurry samples of five selected pig fattening and four broiler farms, and boot swabs from surfaces of fertilized fields (within the last six weeks) around one pig and one broiler farm were inves-tigated for ESBLs/AmpC-producing E. coli. The positive

BUM_3_4_s175-180_12044_Friese_pr 176 26.02.13 10:12


status of these two selected farms was known before sampling. Nine out of all investigated pig farms were tested for MRSA and ESBLs/AmpC-producing E. coli simultaneously.

Analysis of MRSABoot swabs or 25 g faeces were inoculated in 225 ml Mueller Hinton broth (868517, Oxoid Lt., UK) with 6.5% NaCl (MHB+) and homogenized using a stomacher (230 rpm, 2 min). After 24 h of incubation at 37°C 2.5 ml of MHB+ was transferred into 22.5 ml tryptone soy broth (583439, Oxoid Lt., UK) including 75 mg/l aztreonam and 3.5 mg/l cefoxitin (TSB+). This selective broth was incubated for 17 h at 37°C. A loop-full of TSB+ was streaked onto chromogenic MRSA screen agar (CHRO-MagarMRSA™, MAST Diagnostica GmbH, GER) and aerobically incubated at 37°C for 24 h. Characteristic colonies were transferred onto sheep blood agar (Oxoid, CM 0331, GER) and tested for coagulase reaction. Fur-thermore, positive results were confirmed by the detec-tion of the nuc-gene specific for S. aureus and mecA-gene specific for the methicillin resistance (Pasanen et al., 2010). To emphasise the livestock association, exem-plarily, eleven MRSA isolates originating from faeces of pigs, turkey and broilers were tested concerning their belonging to the clonal complex 398 as described before (Stegger et al., 2011).

Analysis of ESBL/AmpC-producing E. coliBoot swabs, 25 g faeces or 25 g slurry were inoculated in 225 ml Luria-Bertani(LB)-broth (Merck kGaA, GER) and aerobically incubated at 37°C for 24 h. A loop-full of LB-broth was streaked onto MacConkey agar plates (Oxoid, CM 0115, Wesel, GER) containing 1 mg/l cefo-taxime and incubated equally. For each sample one characteristic E. coli colony was chosen randomly and the species was confirmed using MALDI TOF (Bruker Daltonik GmbH, Bremen). Subsequently the existence of the N-lactamase genes blaCTX-M, blaTEM, blaSHV and the plasmid-mediated AmpC N-lactamase gene blaCMY-2 was examinded by PCR as described before (Guerra et al., 2001; Zhao et al., 2001; Weill et al., 2004; Batchelor et al., 2005). For isolates in which either the blaTEM and/or the blaSHV gene was detected solely, a disk diffusion test with cefotaxime 30 µg and cefotaxime 30 µg + clavulanic acid 10 µg (D62C, MAST Diagnostica GmbH, Reinfeld, GER) was applied. An increase of the inhibition zone about more than 5 mm in the presence of clavulanic

acid was considered to be indicative for the presence of extended-spectrum N-lactamases.

Results

LaMRSA as well as ESBL/AmpC-producing E. coli could regularly be found in pooled faeces samples of different farm animals. Moreover, a detection of ESBL/AmpC-producing E. coli was possible in samples of slurry and boot swabs of fertilized fields. MRSA was also found in boot swab samples from fertilized fields around three pig farms and one turkey farm.

Pooled faecal samplesTable 1 summarises the detection frequencies of laMRSA and ESBL/AmpC-producing E. coli in pooled faecal sam-ples of different animal farms. LaMRSA could regularly be found (48.6%) in pooled faecal samples of different farms with a positive MRSA-status. Especially in fat-tening pig barns the detection of MRSA was very high (73.3%). In breeding pig farms the resistant pathogen was detected sporadically and significantly less than in fattening farms (b2-test, p = 0.038). Also in samples of turkey and broiler farms the microorganisms could be found although the number of investigated farms is low. The faecal sample of the only sampled MRSA-positive cattle farm was negative. Ten out of eleven (90.9%) tested MRSA isolates originating from the pooled faeces samples belong to the clonal complex 398 and are thus livestock-associated (Stegger et al., 2011). One isolate of a broiler farm was negative for that.

Concerning ESBL/AmpC the majority of pooled faecal samples (60%) was tested positive for ESBL/AmpC-producing E. coli (Tab. 1). In detail, all broiler farms were classified positive for this resistant pathogen. In many dairy cattle farms as well as in breeding pig farms ESBL/AmpC-producing E. coli could be found. In fattening pig farms the detection frequency was lowest (43.8%), however, there was no significant difference between breeding and fattening pig farms (b2-test, p = 0.48). All of the four mentioned N-lactamase genes blaCTX-M, blaTEM, blaSHV or blaCMY-2 could be found in the E. coli isolates with different extent. Among isolates originat-ing from fattening and breeding pig farms the CTX-M-type enzyme was dominant in 14 out of 16 positive isolates whereas five isolates produce both the CTX-M and TEM-type N-lactamases. The remaining two iso-lates from fattening pig farms carried the blaTEM gene. Interestingly, in one of these pig fattening farms also the first carpapenem resistant E. coli from livestock with the blaVIM-1 gene was found in parallel (Fischer et al., 2012). Concerning the eight broiler flocks five isolates had a plasmid-mediated AmpC N-lactamase, three of them in combination with a blaTEM gene. In two isolates only the TEM-type N-lactamase could be detected and in one only the SHV-type. Isolates from cattle farms showed the CTX-M-type of enzyme in two samples, the TEM-type in two, the AmpC-type in one sample and one isolate contained a combination of the three genes blaCTX-M, blaTEM and blaSHV.

Slurry samplesIn all slurry samples (100%) from four broiler farms and five pig fattening farms phenotypic suspected ESBL/AmpC-producing E. coli could be isolated on

TABLE 1: Occurrence of MRSA and ESBL/AmpC-producing E. coli in pooled faeces samples from different animal farms in Germany

MRSA1 in faeces in % ESBL/AmpC- producing E. coli in faeces in %

All farms2 48.6 (18/37) 60 (30/50)Fattening pig farms 73.3 (11/15) 43.8 (7/16)Breeding pig farms3 33.3 (4/12) 56.3 (9/16)Turkey farms 40 (2/5) –Broiler farms 25 (1/4) 100 (8/8)Dairy cattle farms 0 (0/1) 60 (6/10)

The number of positive samples (detection of nuc- and mecA-gene for MRSA, detection of blaCTX-M gene, blaTEM gene, blaSHV gene and/or blaCMY-2 gene for ESBL/AmC-producing E. coli) from the total number of samples is in parenthesis.1 partly published in Friese et al. (2012)2 farms with known positive MRSA status but with unknown ESBL/AmpC status3 including weaner-to-grower farms

BUM_3_4_s175-180_12044_Friese_pr 177 26.02.13 10:12


MacConkey agar. In two samples originating from broiler farms and in two from pig farms the blaTEM gene was detected. In one broiler farm sample the blaCTX-M gene and in another the blaCMY-2 gene was found. In the remaining slurry samples originating from three pig farms none of the four genes was detected by testing only this one randomly chosen E. coli isolate.

Boot swabs of fertilized fieldsBoth MRSA and ESBL/AmpC-producing E. coli could be detected on the surface of previously fertilized fields. MRSA was found around three pig farms. More details concerning these results were shown by Schulz et al. (2012). Phenotypic suspected ESBL/AmpC-producing E. coli occurred on three different locations on a ferti-lized field around one pig farm. In these three samples the blaTEM and blaSHV genes were detected. The blaTEM gene was also found in the isolate of the associated slurry sample of this farm. The environment of the sampled broiler farm showed also positive results for ESBL/AmpC-producing E. coli in the boot swab of the surface of a fertilized field. There the blaSHV gene could be detected.

Disk DiffusionAll isolates with an exclusive detection of blaTEM or blaSHV gene tested in disk diffusion test showed an increase of inhibition zone about more than 5 mm in the presence of clavulanic acid and could be therefore proved as ESBL-producing E. coli.

Discussion

The increasing presence of multiresistant bacteria is a problem in veterinary as well as in human medicine. Especially the occurrence in healthy farm animals is discussed to be a possible source for transmission of laMRSA and ESBL/AmpC-producing Enterobacteriaceae to humans or the environment. This study presents the first results of ongoing research on the spread of multiresistant organisms in the environment of animal barns via the faecal route. To the best of our knowledge this is the first detection of ESBL/AmpC-producing Enterobacteriaceae in faecal samples of healthy pigs, tur-keys and broilers, slurry and the environment of farms in Germany. Our results show a regular detection of MRSA and ESBL/AmpC-producing E. coli in faeces and exemplarily in slurry as well as on fertilized field sur-faces in the vicinity of farms. There were a few isolates suspected to produce ESLB/AmpC in which merely the blaTEM gene or blaSHV could be detected by PCR. This is no evidence for the presence of extended-spectrum N-lactamases. To underline the results of these isolates disk diffusion tests were applied.

The detection of ESBL/AmpC-producing E. coli in pooled faeces samples originating from various animal farms is quite high (43.8% to 100% for the different farm animals) and indicates the importance of the feacal spread of these multiresistant pathogens. Similar results were found by Mesa et al. (2006). This group investi-gated ten broiler and ten pig farms in Spain by testing pooled faecal samples and found the resistant bacteria also in every broiler and in eight pig farms. Furthermore, five examined broiler farms were positive for ESBL-producing E. coli in Italy (Bortolaia et al., 2010) as well

as in Belgium (Smet et al., 2008). However, many other studies in Europe had different study designs focused on either the prevalence of ESBL-producing E. coli in animal samples originating from one individual farm or differ-ent slaughterhouses (Costa et al., 2009; Dierikx et al., 2010; Goncalves et al., 2010). Since our study examined many different farms by investigating one pooled sam-ple the data are hardly comparable. Concerning MRSA, there exist only a few other studies about the detection especially in faeces. Interestingly, samples from fatten-ing farms had a significantly higher MRSA detection rate than those from breeding farms. This phenomenon can also be observed in other sample matrices (Friese et al., 2012) and maybe correlates with a higher animal exchange and the mixing of pigs from different suppliers in fattening farms (Broens et al., 2011). Pletinckx et al. (2011) isolated MRSA in cloacal swabs of broilers from three different farms, Anukool et al. (2011) in a rectal swab of one pig and Szabo et al. (2011) in rectal swabs of experimentally infected swine. Although no manure samples were analysed, the occurrence of viable laMRSA in it is very likely.

However, a few slurry samples were studied exem-plarily for the presence of ESBL/AmpC-producing E. coli and lead to positive results. In addition to this, a detection of these microorganisms as well as MRSA was possible in the direct environment of some selected farms on fertilized fields. A French study investigated cattle farms and their vicinity and led to similar results (Hartmann et al., 2012). They found CTX-M-producing E. coli in faecal samples as well as in soil of the envi-ronment of the barns. Cobbold et al. (2006) detected multiresistant Salmonella in faecal and environmental samples on dairy farms, too. This indicates a potential emission of these multiresistant bacteria in the vicinity of different animal farms. Manure has been applied on the fields which were tested for ESBL/AmpC-producing E. coli in this study within six weeks before our meas-urements. Regarding the fields around the three pig barns investigated for MRSA fertilization within the last three month before our measurements was performed. However, at the time of sampling no residue of manure was visible. In the study of Hartmann et al. (2012) there has not been any application of manure since one year. That all means that MRSA as well as E. coli originat-ing from livestock as a potential carrier of genes for ESBL/AmpC seem to survive in the environment for longer times. Until now there is no specific knowl-edge about the tenacity of MRSA or E. coli on different grounds and under different environmental conditions. Factors like temperature, rain, ultraviolet radia-tion, structure of the soil, concentration of heavy metals or the presence of other microorganisms could influence the viability of these pathogens (Chapple et al., 1992; Wang et al., 2007). Apart from the con-tamination of the surfaces via faeces other routes are conceivable. An emission via the airborne way is possible and has already been described for laMRSA (Friese et al., 2013; Schulz et al., 2012). They detected laMRSA in exhaust air of pig barns. Furthermore, rodents or other wild animals as carriers of laMRSA or ESBL/AmpC-producing E. coli could spread these microorganisms (van de Giessen et al., 2009; Guen-ther et al., 2011). However, a risk for farms, animals or humans living in the vicinity can currently not be estimated.

BUM_3_4_s175-180_12044_Friese_pr 178 26.02.13 10:12


Acknowledgement

This work was financially supported by the German Fed-eral Ministry of Food, Agriculture and Consumer Protec-tion (BMELV) through the Federal Office for Agriculture and Food (BLE, funding number 2808HS029/2808HS040) as well as the Federal Ministry of Education and Research (funding number 01Kl1013C). We thank Dr. Beatriz Guerra for scientific advices. All cooperat-ing farmers and veterinarians as well as the excellent technical assistance of H. Jansen, M. Thieck, S. Sellen-thin and K. Fiedler (Institute for Animal Hygiene and Environmental Health) are gratefully acknowledged.

Conflict of interest: There are no protected financial, professional or other personal interests in any product, service and/or a company that could affect in this manu-script presented content or opinions.

References

Anukool U, O’Neill CE, Butr-Indr B, Hawkey PM, Gaze WH, Wellington EM (2011): Meticillin-resistant Staphylococcus aureus in pigs from Thailand. Int J Antimicrob Agents 38: 86–87.

Batchelor M, Hopkins K, Threlfall EJ, Clifton-Hadley FA, Stall-wood AD, Davies RH, Liebana E (2005): bla(CTX-M) genes in clinical Salmonella isolates recovered from humans in England and Wales from 1992 to 2003. Antimicrob Agents Chemother 49: 1319–1322.

Bortolaia V, Guardabassi L, Trevisani M, Bisgaard M, Venturi L, Bojesen AM (2010): High diversity of extended-spectrum beta-lactamases in Escherichia coli isolates from Italian broiler flocks. Antimicrob Agents Chemother 54: 1623–1626.

Brinas L, Moreno M A, Teshager T, Zarazaga M, Saenz Y, Porrero C, Dominguez L, Torres C (2003): Beta-lactamase characteriza-tion in Escherichia coli isolates with diminished susceptibility or resistance to extended-spectrum cephalosporins recovered from sick animals in Spain. Microb drug resis 9: 201–209.

Broens EM, Graat EA, van der Wolf P J, van de Giessen AW, van Duijkeren E, Wagenaar JA, van Nes A, Mevius DJ, de Jong MC (2011): MRSA CC398 in the pig production chain. Prev Vet Med 98: 182–189.

Chapple RM, Inglis B, Stewart PR (1992): Lethal and mutational effects of solar and UV radiation on Staphylococcus aureus. Arch microbiol 157: 242–248.

Cobbold RN, Rice DH, Davis MA, Besser T E, Hancock DD (2006): Long-term persistence of multi-drug-resistant Salmo-nella enterica serovar Newport in two dairy herds. J Am Vet Med Assoc 228: 585–591.

Coque TM, Baquero F, Canton R (2008): Increasing prevalence of ESBL-producing Enterobacteriaceae in Europe. Euro Surveill 13.

Costa D, Vinue L, Poeta P, Coelho AC, Matos M, Saenz Y, Somalo S, Zarazaga M, Rodrigues J, Torres C (2009): Prevalence of extended-spectrum beta-lactamase-producing Escherichia coli isolates in faecal samples of broilers. Vet Microbiol 138: 339–344.

Cuny C, Nathaus R, Layer F, Strommenger B, Altmann D, Witte W (2009): Nasal colonization of humans with methicillin-resistant Staphylococcus aureus (MRSA) CC398 with and without exposure to pigs. PLoS One 4: e6800.

Dierikx C, van Essen-Zandbergen A, Veldman K, Smith H, Mevius D (2010): Increased detection of extended spectrum beta-lactamase producing Salmonella enterica and Escherichia coli isolates from poultry. Vet Microbiol 145: 273–278.

Ewers C, Bethe A, Semmler T, Guenther S, Wieler LH (2012): Extended-spectrum beta-lactamase-producing and AmpC-pro-ducing Escherichia coli from livestock and companion animals, and their putative impact on public health: a global perspective. Clin Microbiol Infect 18: 646–655.

Fischer J, Rodriguez I, Schmoger S, Friese A, Roesler U, Hel-muth R, Guerra B (2012): Escherichia coli producing VIM-1 carbapenemase isolated on a pig farm. J Antimicrob Chemother 67: 1793–1795.

Friese A, Schulz J, Hoehle L, Fetsch A, Tenhagen BA, Hartung J, Roesler U (2012): Occurrence of MRSA in air and housing environment of pig barns. Vet Microbiol 158: 129–135.

Friese A, Schulz J, Zimmermann K, Tenhagen BA, Fetsch A, Hartung J, Rösler U (2013): The occurrence of LA-MRSA in turkey and broiler barns and the contamination of air and soil surfaces in their vicinity. Appl Environ Microbiol (accepted with minor revision).

Goncalves A, Torres C, Silva N, Carneiro C, Radhouani H, Coelho C, Araujo C, Rodrigues J, Vinue L, Somalo S, Poeta P, Igrejas G (2010): Genetic characterization of extended-spectrum beta-lactamases in Escherichia coli isolates of pigs from a Portuguese intensive swine farm. Foodborne Pathog Dis 7: 1569–1573.

Graveland H, Wagenaar JA, Bergs K, Heesterbeek H, Heederik D (2011): Persistence of livestock associated MRSA CC398 in humans is dependent on intensity of animal contact. PLoS One 6: e16830.

Guenther S, Ewers C, Wieler L H (2011): Extended-spectrum beta-lactamases producing E. coli in wildlife, yet another form of environmental pollution? Front Microbiol 2: 246.

Guerra B, Soto SM, Arguelles JM, Mendoza MC (2001): Multi-drug resistance is mediated by large plasmids carrying a class 1 integron in the emergent Salmonella enterica serotype [4,5,12:i:-]. Antimicrob Agents Chemother 45: 1305–1308.

Hartmann A, Locatelli A, Amoureux L, Depret G, Jolivet C, Gueneau E, Neuwirth C (2012): Occurrence of CTX-M Pro-ducing Escherichia coli in Soils, Cattle, and Farm Environment in France (Burgundy Region). Front Microbiol 3: 83.

Liebana E, Batchelor M, Hopkins KL, Clifton-Hadley FA, Teale CJ, Foster A, Barker L, Threlfall EJ, Davies RH (2006): Lon-gitudinal farm study of extended-spectrum beta-lactamase-mediated resistance. J Clin Microbiol 44: 1630–1634.

Meemken D, Cuny C, Witte W, Eichler U, Staudt R, Blaha T (2008): Occurrence of MRSA in pigs and in humans involved in pig production – preliminary results of a study in the northwest of Germany. Dtsch Tierarztl Wochenschr 115: 132–139.

Mesa RJ, Blanc V, Blanch AR, Cortes P, Gonzalez JJ, Lavilla S, Miro E, Muniesa M, Saco M, Tortola MT, Mirelis B, Coll P, Llagostera M, Prats G, Navarro F (2006): Extended-spectrum beta-lactamase-producing Enterobacteriaceae in different envi-ronments (humans, food, animal farms and sewage). J Antimi-crob Chemother 58: 211–215.

Overdevest I, Willemsen I, Rijnsburger M, Eustace A, Xu, L, Hawkey P, Heck M, Savelkoul P, Vandenbroucke-Grauls C, van der Zwaluw K, Huijsdens X, Kluytmans J (2011): Extended-spectrum beta-lactamase genes of Escherichia coli in chicken meat and humans, The Netherlands. Emerg Infect Dis 17:1216–1222.

Pasanen T, Korkeila M, Mero S, Tarkka E, Piiparinen H, Vuopio-Varkila J, Vaara M, Tissari P (2010): A selective broth enrich-ment combined with real-time nuc-mecA-PCR in the exclusion of MRSA. Apmis 118: 74–80.

Pletinckx LJ, Verhegghe M, Dewulf J, Crombe F, de Bleecker Y, Rasschaert G, Goddeeris BM, de Man I (2011): Screening of poultry-pig farms for methicillin-resistant Staphylococcus aureus: sampling methodology and within herd prevalence in broiler flocks and pigs. Infect Genet Evol 11: 2133–2137.

Richter A, Sting R, Popp C, Rau J, Tenhagen BA, Guerra B, Hafez HM, Fetsch A (2012): Prevalence of types of methicillin-resistant Staphylococcus aureus in turkey flocks and personnel attending the animals. Epidemiol Infect 140: 2223–2232.

BUM_3_4_s175-180_12044_Friese_pr 179 26.02.13 10:12


Rodriguez I, Barownick W, Helmuth R, Mendoza MC, Rodicio MR, Schroeter A, Guerra B (2009): Extended-spectrum {beta}-lactamases and AmpC {beta}-lactamases in ceftiofur-resistant Salmonella enterica isolates from food and livestock obtained in Germany during 2003–07. J Antimicrob Chemother 64: 301–309.

Schulz J, Friese A, Klees S, Tenhagen BA, Fetsch A, Rosler U, Hartung J (2012): Longitudinal Study of the Contamination of Air and of Soil Surfaces in the Vicinity of Pig Barns by Livestock-Associated Methicillin-Resistant Staphylococcus aureus. Appl Environ Microbiol 78: 5666–5671.

Smet A, Martel A, Persoons D, Dewulf J, Heyndrickx M, Catry B, Herman L, Haesebrouck F, Butaye P (2008): Diversity of extended-spectrum beta-lactamases and class C beta-lacta-mases among cloacal Escherichia coli Isolates in Belgian broiler farms. Antimicrob Agents Chemother 52: 1238–1243.

Smet A, Martel A, Persoons D, Dewulf J, Heyndrickx M, Her-man L, Haesebrouck F, Butaye P (2009): Broad-spectrum beta-lactamases among Enterobacteriaceae of animal origin: molecular aspects, mobility and impact on public health. FEMS Microbiol Rev 34: 295–316.

Spohr M, Rau J, Friedrich A, Klittich G, Fetsch A, Guerra B, Hammerl JA, Tenhagen BA (2011): Methicillin-resistantStaphylococcus aureus (MRSA) in three dairy herds in southwest Germany. Zoonoses Public Health 58: 252–261.

Stegger M, Lindsay JA, Moodley A, Skov R, Broens EM, Guardabassi L (2011): Rapid PCR detection of Staphylococcus aureus clonal complex 398 by targeting the restriction-modifica-tion system carrying sau1-hsdS1. J Clin Microbiol 49: 732–734.

Szabo I, Beck B, Friese A, Fetsch A, Tenhagen BA, Roesler U (2011): Colonization Kinetics of Different Methicillin-Resistant Staphylococcus aureus Sequence Types in Pigs and Host Suscep-tibilities. Appl Environ Microbiol 78: 541–548.

Van de Giessen AW, van Santen-Verheuvel MG, Hengeveld PD, Bosch T, Broens EM, Reusken CB (2009): Occurrence of methicillin-resistant Staphylococcus aureus in rats living on pig farms. Prev Vet Med 91: 270–273.

Wang Y, Shi J, Wang H, Lin Q, Chen X, Chen Y (2007): The influ-ence of soil heavy metals pollution on soil microbial biomass, enzyme activity, and community composition near a copper smelter. Ecotoxicol Environl Saf 67: 75–81.

Weill FX, Demartin M, Tande D, Espie E, Rakotoarivony I, Grimont PA (2004): SHV-12-like extended-spectrum-beta-lactamase-producing strains of Salmonella enterica serotypes Babelsberg and Enteritidis isolated in France among infants adopted from Mali. J Clin Microbiol 42: 2432–2437.

Wieler LH, Semmler T, Eichhorn I, Antao EM, Kinnemann B, Geue L, Karch H, Guenther S, Bethe A (2011a): No evidence of the Shiga toxin-producing E. coli O104:H4 outbreak strain or enteroaggregative E. coli (EAEC) found in cattle faeces in northern Germany, the hotspot of the 2011 HUS outbreak area. Gut Pathog 3: 17.

Wieler LH, Ewers C, Guenther S, Walther B, Lubke-Becker A (2011b): Methicillin-resistant staphylococci (MRS) and extended-spectrum beta-lactamases (ESBL)-producing Entero-bacteriaceae in companion animals: nosocomial infections as one reason for the rising prevalence of these potential zoonotic pathogens in clinical samples. Int J Med Microbiol 301: 635–641.

Zhao S, White DG, McDermott PF, Friedman S, English L, Ayers S, Meng J, Maurer JJ, Holland R, Walker RD (2001): Identi-fication and expression of cephamycinase bla(CMY) genes in Escherichia coli and Salmonella isolates from food animals and ground meat. Antimicrob Agents Chemother 45: 3647–3650.

Address for correspondence:Dr. Anika FrieseFreie Universität BerlinInstitut für Tier- und UmwelthygieneRobert-von-Osterag-Straße 1–1314163 [email protected]

DanksagungenDie Schlütersche Verlagsgesellschaft & Co. KG dankt Herrn Professor Hartung herzlich für die langjährige Zusam-menarbeit. Die Schriftleitung der Zeitschrift „Deutsche Tierärzliche Wochenschrift" (DTW) lag viele Jahre in seinen vertrauensvollen Händen.

Der Verlag und die Redaktion wünschen Professor Hartung und seiner Ehefrau eine glückliche Zeit im zukünftigen wohlverdienten Ruhestand.

Die Initiatoren und die Autorenschaft dieses Themenheftes bedanken sich ganz herzlich für die finanzielle Unterstützung, ohne die eine vollständige Open-Access-Veröffentlichung der Arbeiten dieser Ausgabe einschließlich des Editorials nicht möglich gewesen wäre.

Unser besonderer Dank gilt:

Boehringer Ingelheim Virbac Tierarzneimittel Vetmedica GmbH GmbH

Hölscher + Leuschner BfT Bundesverband GmbH & Co. KG für Tiergesundheit e. V.

BUM_3_4_s175-180_12044_Friese_pr 180 26.02.13 10:12

open access faecal occurrence and emissions of live- stock

Documents