sign in sign up
<<
Home , Cefapirin, Cefquinome, Ceftiofur

P 1103 Antimicrobial susceptibility of mastitis pathogens isolated from diseased dairy cows across Europe: VetPath monitoring results

V. Thomas1, A. de Jong1, S. Simjee1, K. Maher2, I. Morrissey2, 22nd ECCMID P. Butty1, U. Klein1, H. Marion1, H. Moyaert1, D. Rigaut1, M. Vallé1 London, United Kingdom Contact: 31 March – 3 April 2012 1VetPath Study Group, Brussels, Belgium; 2Quotient Bioresearch, Fordham, UK [email protected]

Introduction Results and discussion VetPath is of concern for the antimicrobial therapy of both hu- In total 1066 isolates were recovered during the mans and animals. Monitoring of antimicrobial susceptibility trends over time collection period, 363 E. coli, 356 S. aureus and is important to ensure long-term antimicrobial efficacy. Validated, harmonized 347 S. uberis. The MIC parameters and percent- l The VetPath project is a pan-European pro- and continuous monitoring of antimicrobial susceptibility of bacterial pathogens ages of resistance are presented in table 1 and 2 gramme dedicated to the collection and to the is of importance for a science based risk assessment on resistance. respectively. monitoring of susceptibility of veteri- nary pathogens from diseased food-producing The VetPath programme collects bacteria from diseased food animals (cattle, For E. coli, the highest activity was observed for one animals not recently exposed to antimicrobials. swine, poultry) across Europe, employing a protocol with harmonized methods beta-lactam, and two fluoroquinolones, l VetPath is an initiative of, and is coordinated of sampling and bacterial isolation. A central laboratory (Quotient Bioresearch, enrofloxacin and marbofloxacin with an MIC50 of by, the European Animal Health Study Center UK) conducts MIC determinations using a panel of licensed antimicrobials com- 0.03 mg/L and MIC90 of 0.03 to 0.06 mg/L. (CEESA) in Brussels. It has been operational monly used in veterinary medicine. The present surveillance study is part of MIC90 was 0.5 mg/L. / and since 2000. CEESA’s membership is composed the on-going VetPath programme co-ordinated by the European Animal Health cephapirin showed comparable activity with MIC90 of international pharmaceutical companies re- Study Centre (CEESA). of 8 mg/L. Cephalexin MIC90 was slightly higher at 32 mg/L. Very few have a CLSI break- searching and producing veterinary medicinal Three major bacterial pathogens of acute mastitis, , Staphylo- point and from Table 2 it can be concluded that products. coccus aureus and uberis1, were recovered in 8 EU countries no resistance was observed for ceftiofur and only l Sponsors of the current VetPath project were between 2002 and 2006. The results of their antibiotic susceptibility are pre- 1% resistance for amoxicillin/clavulanic acid. Resis- the following companies: Bayer Animal Health sented here. GmbH, Germany; CEVA Sante Animale, France; tance to tetracycline was moderate at 16%. Two % Elanco Animal Health, UK; MSD Animal Health, of the isolates showed intermediate susceptibility The Netherlands; Novartis Animal Health Inc., Methods to amoxicillin/clavulanic acid and 0.5% to tetracy- cline. For neomycin, although the MIC was 4 mg/L, Switzerland; Pfizer Animal Health, Belgium; Ve- • Eight European countries (Belgium, Czech Republic, France, Germany, Ire- 90 toquinol S. A., France; ­Virbac, France. 5 isolates (1.4%) were found with an MIC > 64 mg/L. land, Italy, The Netherlands, and The United Kingdom) were involved in the l The culture collection of VetPath was organized study. Milk samples were collected from cows with acute clinical signs, not Findings for E. coli are similar to those reported by by CEESA, and subsequently from each partici- recently exposed to antimicrobial treatment. Samples were processed ac- GERM-Vet, the German national surveillance pro- pating country transmitted to a central Europe- cording to standard procedures for the isolation of major bacterial pathogens gramme and MARAN, the surveillance programme an laboratory (Quotient Bioresearch, Fordham, at regional laboratories. Only one isolate per farm was selected. in the Netherlands.3,4 UK). All antimicrobial susceptibility determina- tions were performed by Quotient Bioresearch. • Isolates were transferred to the central laboratory for broth microdilution MIC With regards to S. aureus, most antibiotics showed determination according to the Clinical and Laboratory Standard Institute comparable activity with MIC50 in the range of 0.12 l The CEESA project manager is Hervé Marion,­ 2 guidelines using 96 well TREK plates. Nine antibiotics were tested against to 1 mg/L and MIC90 of 0.25 to 1 mg/L with the ex- CEESA, Rue Defacqz 1, Bte 8, B-1000 Brussels, E. coli with the following ranges: amoxicillin/clavulanic acid (0.06/0.03-64/ ception of cephalexin which showed slightly higher Belgium. 32 mg/L), cefquinome (0.002-8 mg/L), ceftiofur (0.002-8 mg/L), cephalexin MIC50 and MIC90 at 2 and 4 mg/L and for For correspondence: which a low MIC of 0.03 mg/L was observed but Tel: +32 2 543 7572 (0.03-32 mg/L), cephapirin (0.03-32 mg/L), enrofloxacin (0.002-16 mg/L), 50 a MIC of 2 mg/L corresponding to 38% of resis- e-mail: [email protected] ­marbofloxacin (0.002-16 mg/L), neomycin (0.12-64 mg/L) tetracycline 90 (0.06-64 mg/L. Three additional antibiotics were tested against S. aureus and tance. Resistance to tetracycline was low at 7.3% S. uberis; they were (0.03-8 mg/L), penicillin (0.004-8 mg/L) and and for erythromycin 1.4% of the isolates were erythromycin (0.03-32 mg/L). resistant, 0.3% showed intermediate susceptibil- ity. No resistance was observed for ceftiofur and • The following quality control strains were used: faecalis ATCC 29212, amoxicillin/clavulanic acid. Abstract E. coli ATCC 25922, E. coli ATCC 35218, ATCC 27853, A. pleuropneumoniae ATCC 27090, S. aureus ATCC 25923, S. aureus ATCC 29213, Finally for S. uberis, MIC90 in the range of 0.25 to Objectives: VetPath is an ongoing pan-European resis- and S. pneumoniae ATCC 49619. The numbers of isolates included for suscep- 0.5 mg/L was observed for penicillin, amoxicillin/cla- tance monitoring program for veterinary pathogens iso- tibility testing are detailed in Tables 1 and 2. MIC50 and MIC90 values were deter- vulanic acid, cefquinome, and cephalexin. lated from diseased antimicrobial-naive cattle, pigs and mined, and resistance was calculated for each drug and organism according to For ceftiofur as well as for the fluoroquinolones enro- 2 poultry. Antimicrobial susceptibilities of mastitis patho- CLSI veterinary breakpoints where available (Table 2). floxacin and marbofloxacin MIC90 values were in the gens isolated from lactating cattle are presented here. range of 1 to 2 mg/L. High MIC90 i.e. 32 mg/L and Methods: Milk was sampled from dairy cows with acute Table 1. MIC parameters (mg/L) for different antibiotics against bacte- above were found for erythromycin (19% resistance), mastitis, not recently exposed to antimicrobial treatment, ria isolated from mastitis in dairy cows in eight EU countries neomycin (no resistance breakpoint) and tetracycline (no resistance breakpoint). While no resistance was in 8 EU countries, mainly during 2002-2006. Escherichia MIC parameters (mg/L) coli (Ec), aureus (Sa) and Streptococcus observed for penicillin, amoxicillin/clavulanic acid E. coli S. aureus S. uberis and ceftiofur, 28.2% of the isolates showed inter- uberis (Su), were isolated and biochemically identified (n=363) (n=356) (n=347) (one isolate per farm). MIC values of 9 (Ec) and 12 (Sa, mediate susceptibility to penicillin but only 0.6% to MIC50 MIC90 MIC50 MIC90 MIC50 MIC90 Su) antibiotics were determined in a central laboratory by ceftiofur and 1.1% to erythromycin. broth microdilution as per CLSI guidelines. Susceptibil- Penicillin - - 0.03 2 0.03 0.25 Cloxacillin - - 0.25 0.5 0.5 4 ity was expressed as MIC50 and MIC90, and percentage Conclusions resistance was determined for antibiotics with defined Amoxicillin/ 4 8 0.25 1 0.12 0.5 CLSI breakpoints (M31-A3, 2008). clavulanic acid The results of this pan-European survey, with standardized methods, in general show an ab- Results: In total, 1066 isolates were recovered, 363 Ec, 356 Sa Cefquinome 0.03 0.06 0.5 0.5 0.03 0.25 sence or a low prevalence of antimicrobial re- and 347 Su. For Ec, resistance to beta-lactam antibiotics was Ceftiofur 0.25 0.5 1 1 0.12 2 sistance among the major pathogens collected virtually absent; tetracycline resistance amounted to 16%. For Cephalexin 4 8 2 4 0.5 0.5 the tested without CLSI breakpoints, MIC from non-treated dairy cows with acute mas- 90 Cephapirin 8 32 0.12 0.25 0.06 0.5 amounted to 0.06 mg/L for cefquinome and 8 and 64 mg/L for titis. Some exceptions noted were tetracycline cephalexin and cephapirin. MIC distributions of marbofloxa- Enrofloxacin 0.03 0.03 0.12 0.25 0.5 1 vs. E. coli (16% resistance), penicillin vs. S. au- cin and enrofloxacin were similar, with MIC50 and MIC90 of 0.03 Marbofloxacin 0.03 0.06 0.25 0.5 1 2 reus (38% resistance) and erythromycin vs. and 0.03-0.06 mg/L. For neomycin, five Ec isolates showed Erythromycin - - 0.25 0.5 0.06 ≥64 S. uberis (19% resistance). The VetPath network, MICs > 128 mg/L, while MIC50 was 1 mg/L. For Sa, resistance to amoxicillin/clavulanic acid was absent, whilst penicillin re- Neomycin 1 4 0.5 1 64 ≥128 based on quantitative and representative data, sistance was 38%, but MIC90 for cloxacillin was as low as Tetracycline 2 64 0.25 0.5 0.25 32 enables to reliably monitor antimicrobial sus- 0.5 mg/L. MIC90 of cefquinome and cephapirin were 0.25 and - : not tested ceptibility of major veterinary pathogens across 0.5 mg/L; for cephalexin 4 mg/L. Erythromycin and tetracy- Europe. cline resistance were low (1.4 and 7.3%, respectively). MIC 50/90s Table 2. Prevalence of antibiotic resistance (%) in bacteria isolated of enrofloxacin and marbofloxacin varied from 0.12 to 0.5 from mastitis applying CLSI interpretive criteria mg/L. Neomycin MICs ranged from 0.25 to 2 mg/L, except for References Percentage of resistance two isolates. In contrast, for Su beta-lactam resistance was 1. Cressier B, Bissonette N. (2011). Assessment of an extraction pro- absent, whereas erythromycin resistance was 19%. For the Antimicrobial Resistance ­breakpoint E.coli (Ec) S.aureus (Sa) S.uberis (Su) tocol to detect the major mastitis-causing pathogens in bovine milk. J. Dairy Sci 94: 2171-84. beta-lactam antibiotics without CLSI breakpoints, MIC90 of (CLSI) (n=363) (n=356) (n=347) cefquinome, cephalexin and cephapirin were 0.25-0.5 mg/L; Amoxicillin/clavulanic acid Ec, Su: R ≥ 32 mg/L 1.0 0.0 0.0 2. CLSI. (2008). Performance standards for antimicrobial disk and dilu- Sa: R ≥ 8mg/L tion susceptibility tests for bacteria isolated from animals; Approved cloxacillin MIC90 was 4 mg/L. Fluoroquinolone MIC50/90s were Standard-third edition. M31-A3. Clinical and Laboratory Standards 0.5-2 mg/L, whereas neomycin MIC50 and MIC90 were 64 Ceftiofur R ≥ 8mg/L 0.0 0.0 0.0 Institute, Wayne, PA, USA. and ≥ 128 mg/L. Erythromycin Sa ≥ 8 mg/L - 1.4 19.0 3. GERMAP 2008. (2009). Bericht über den Antibiotikaverbrauch und Su ≥ 1 mg/L die Verbreitung von Antibiotikaresistenzen in der Human-und Veter- Conclusions: Overall, the prevalence of antibiotic resis- inärmedizin in Deutschland. Report on the usage of antibiotics and tance is low among the three major mastitis pathogens Penicillin Sa ≥0.25 mg/L - 38.0 0.0 the development of resistance in human and veterinary medicine. ISBN 978-3-00-025097-2. isolated from diseased but non-treated dairy cows across Su ≥4 mg/L Tetracycline Ec, Sa ≥ 16 mg/L 16.0 7.3 - 4. MARAN 2008. (2009). Monitoring of Antimicrobial Resistance and the EU, however penicillin resistance of Sa is fairly high. Antibiotic Usage in Animals in the Netherlands in 2008. www.maran. Generally, the tested antibiotics exhibited high efficacy. - : not tested wur.nl.