Antimicrobial susceptibility among respiratory tract pathogens isolated from diseased cattle and pigs from different parts of Europe.

AIMS: To survey antibiotic susceptibility of bacteria causing cattle and pig respiratory infections in 10 European countries. METHODS AND RESULTS: Non-replicate nasopharyngeal/nasal or lung swabs were collected from animals with acute respiratory signs during 2015-2016. Pasteurella multocida, Mannheimia haemolytica, Histophilus somni from cattle (n = 281), and P. multocida, Actinobacillus pleuropneumoniae, Glaesserella parasuis, Bordetella bronchiseptica, and Streptococcus suis from pigs (n = 593) were isolated. MICs were assessed following CLSI standards and interpreted using veterinary breakpoints where available. Histophilus somni isolates were fully antibiotic susceptible. Bovine P. multocida and M. haemolytica were susceptible to all antibiotics, except tetracycline (11.6%-17.6% resistance). Low macrolide and spectinomycin resistance was observed for P. multocida and M. haemolytica (1.3%-8.8%). Similar susceptibility was observed in pigs, where breakpoints are available. Resistance in P. multocida, A. pleuropneumoniae, and S. suis to ceftiofur, enrofloxacin, and florfenicol was absent or <5%. Tetracycline resistance varied from 10.6% to 21.3%, but was 82.4% in S. suis. Overall multidrug-resistance was low. Antibiotic resistance in 2015-2016 remained similar as in 2009-2012. CONCLUSIONS: Low antibiotic resistance was observed among respiratory tract pathogens, except for tetracycline.

European-wide antimicrobial resistance monitoring in commensal Escherichia coli isolated from healthy food animals between 2004 and 2018.

OBJECTIVES: To describe the susceptibility of Escherichia coli to medically important antibiotics, collected over four periods (2004-2006, 2008-2009, 2013-2014, 2017-2018), from food-producing animals at slaughter. METHODS: Intestinal contents from cattle, pigs and broilers were randomly sampled (5-6 countries/host; ≥4 abattoirs/country; one sample/animal/farm) for isolation of Escherichia coli; antimicrobial susceptibilities were centrally determined by CLSI agar dilution. Clinical breakpoints (CLSI) and epidemiological cut-off values (EUCAST) were applied for data interpretation. RESULTS: In total, 10 613 E. coli strains were recovered. In broilers, resistance percentages were the lowest (P ≤ 0.01) in the latest time period. A significant decrease in MDR over time was also observed for broilers and a tendency for a decrease for pigs. Resistance to meropenem and tigecycline was absent, and resistance to azithromycin was 0.2%-2.0%. Also, low resistance to third-generation cephalosporins (1.1%-7.4%) was detected in broilers. Resistance to colistin varied between 0.1%-4.8%. E. coli from broilers showed high resistance to ciprofloxacin (7.3%-23.3%), whereas for cattle and pigs this was 0.2%-2.5%. Low/moderate resistance to chloramphenicol (9.3%-21.3%) and gentamicin (0.9%-7.0%) was observed in pigs and broilers. The highest resistance was noted for ampicillin (32.7%-65.3%), tetracycline (41.3%-67.5%), trimethoprim (32.0%-35.7%) and trimethoprim/sulfamethoxazole (27.5%-49.7%) from pigs and broilers, with marked country differences. MDR peaked in pigs and broilers with 24 and 26 phenotypes, with 21.9%-26.2% and 18.7%-34.1% resistance, respectively. CONCLUSIONS: In this pan-EU survey antibiotic susceptibility of commensal E. coli varied largely between antibiotics, animal species and countries. Resistance to critically important antibiotics for human medicine was absent or low, except for ciprofloxacin in broilers and ampicillin in pigs and broilers.

Minimal inhibitory concentration of seven antimicrobials to Mycoplasma gallisepticum and Mycoplasma synoviae isolates from six European countries.

Mycoplasma gallisepticum and Mycoplasma synoviae are bacterial pathogens that cause disease in poultry, adversely affecting their health and welfare, and are a financial burden on producers. This manuscript describes the results of the MycoPath project that is the first international antimicrobial susceptibility programme for mycoplasma pathogens isolated from poultry. Improved comparative analysis of minimal inhibitory concentration (MIC) results from participating countries was facilitated by using one laboratory determining all MICs. Chicken and turkey isolates were obtained from France, Germany, Great Britain, Hungary, Italy and Spain during 2014-2016. One isolate per farm was retained. The MIC of seven antimicrobial agents was determined using a broth microdilution method, with Friis Medium (M. gallisepticum) or Modified Chanock's Medium (M. synoviae). Of the 222 isolates recovered, 82 were M. gallisepticum and 130 were M. synoviae. M. gallisepticum MIC50/90 values were 0.12/0.5, 2/8, 0.5/4, 0.12/>64, 0.008/0.062, 0.008/32, 0.062/4 mg/l for doxycycline, enrofloxacin, oxytetracycline, spiramycin, tiamulin, tilmicosin and tylosin, respectively. For M. synoviae, the values were 0.5/1, 8/16, 0.5/1, 0.5/8, 0.25/0.5, 0.062/2 and 0.062/16 mg/l respectively. A bimodal MIC distribution for the fluoroquinolone (enrofloxacin) and the macrolides (spiramycin, tilmicosin and tylosin) indicate that both species have sub-populations that are less susceptible in vitro to those antimicrobials. Some differences in susceptibilities were observed according to host species, Mycoplasma species, and country of origin. This study provides a baseline of novel data for future monitoring of antimicrobial resistance in poultry Mycoplasma species. Additionally, this information will facilitate the selection of the antimicrobial agents most likely to be effective, thus ensuring their minimal use with targeted and correct therapeutic treatments.Highlights First large-scale pan-European collection of representative Mg and Ms isolates.MIC values assessed in central laboratory for Mg and Ms from chickens and turkeys.Range of MIC values for 82 Mg and 130 Ms isolates to seven licenced antibiotics shown.Data can be used to help determine Mg and Ms veterinary-specific breakpoints.

Antimicrobial susceptibility monitoring of canine and feline skin and ear pathogens isolated from European veterinary clinics: results of the ComPath Surveillance programme.

BACKGROUND: The ComPath project is a pan-European programme dedicated to the monitoring of antimicrobial susceptibility of canine and feline pathogens using standardized methods and centralized minimal inhibitory concentration (MIC) determination. OBJECTIVES: To report antimicrobial susceptibilities of major pathogens isolated from nontreated animals with acute clinical signs of skin, wound or ear infections in 2013-2014. METHODS AND MATERIALS: MICs were determined by agar dilution for commonly used drugs and interpreted using Clinical and Laboratory Standards Institute (CLSI) breakpoints, if available. RESULTS: Of 1,676 isolates recovered, the main species isolated from dogs were Staphylococcus pseudintermedius, followed by Streptococcus spp., Pseudomonas aeruginosa and Escherichia coli. In cats, Pasteurella multocida, coagulase-negative staphylococci (CoNS) and Staphylococcus aureus were isolated most frequently. Resistance rates observed for S. pseudintermedius were <26.7% for penicillin, clindamycin and chloramphenicol, and ≤11.5% for ampicillin, amoxicillin/clavulanate, cefalexin, cefovecin, gentamicin and fluoroquinolones. For S. aureus, resistance rates ranged up to 90.9% for ß-lactams, and were 19.7% for clindamycin, 27% for fluoroquinolones and 0.0-6.1% for other drugs. The mecA gene was confirmed by PCR in 10.6% of S. pseudintermedius, 11.6% of CoNS and 31.4% of S. aureus isolates. In streptococci/enterococci, resistance to penicillin, ampicillin and chloramphenicol ranged from 0.0% to 11.3%, whereas fluoroquinolone resistance ranged from 0.0% to 8.5%. For E. coli, resistance ranged from 13.8 to 15.9% for fluoroquinolones and from 86.2% to 100.0% for ß-lactams. Low rates of resistance (0.0-6.3%) were observed in P. multocida, and for P. aeruginosa resistance to gentamicin was 10.3%. CONCLUSION: Overall, antimicrobial resistance of cutaneous/otic pathogens isolated from dogs and cats was low (1-10%) to moderate (10-20%). For several pathogens, the paucity of CLSI recommended breakpoints for veterinary use is a bottleneck.

Antimicrobial resistance monitoring in commensal enterococci from healthy cattle, pigs and chickens across Europe during 2004-14 (EASSA Study).

OBJECTIVES: The European Antimicrobial Susceptibility Surveillance in Animals (EASSA) programme collects zoonotic and commensal bacteria from healthy food-producing animals at slaughter and tracks their susceptibility to medically important antibiotics. Results for enterococci, collected over three time periods, are presented. METHODS: Intestinal contents from cattle, pigs and chickens were randomly sampled (five or six countries/host; at least four abattoirs/country; one sample/animal/farm) for isolation of enterococci; antimicrobial susceptibilities were centrally assessed by CLSI agar dilution. Clinical breakpoints (CLSI) and epidemiological cut-off values (EUCAST) were applied for data interpretation. RESULTS: In total, 2435 Enterococcus faecium and 1389 Enterococcus faecalis strains were recovered. Seven E. faecium/faecalis strains were linezolid resistant. One E. faecium strain was non-WT (NWT), with a daptomycin MIC of 8 mg/L. Clinical vancomycin resistance was very low or absent; eight strains had decreased susceptibility (MICs of 8 mg/L). Two strains were clinically resistant to tigecycline. Little resistance to ampicillin or gentamicin was observed. Clinical resistance of E. faecium to quinupristin/dalfopristin was slightly higher (2.2%-33.6%) and 38.5%-83.2% of the strains were classified NWT. Very high resistance to tetracycline (67.4%-79.1%) and erythromycin (27.1%-57.0%) was noted for E. faecium and E. faecalis in pigs and chickens. For both of these compounds, similar NWT results were observed for Enterococcus hirae (n = 935), Enterococcus durans (n = 286) and Enterococcus casseliflavus (n = 154) whereas the percentage of NWT for linezolid, tigecycline and vancomycin was generally zero or low. CONCLUSIONS: In this pan-EU survey of commensal enterococci, antibiotic susceptibility varied widely between antibiotics, animal species, countries and enterococcal species. Clinical resistance to antibiotics that are critically important for human medicine was absent or low, except for erythromycin.

Antimicrobial Susceptibility of Canine and Feline Urinary Tract Infection Pathogens Isolated from Animals with Clinical Signs in European Veterinary Practices during the Period 2013-2018.

Bacterial urinary tract infections (UTIs) occur frequently in companion animals and are often treated with antibiotics. However, antimicrobial resistance can severely hamper treatment success. Therefore, antimicrobial susceptibility monitoring is key. UTI isolates were obtained from dogs and cats in two collection periods (ComPath II: 2013-2014 and ComPath III: 2017-2018) as part of CEESA's ComPath programme. Susceptibility testing of the UTI isolates (2021 in total) was carried out at one central laboratory using agar and broth dilution methodology as recommended by the Clinical and Laboratory Standards Institute. Escherichia coli was the most frequently isolated bacterium in UTI in both dogs (46.9%, 43.1%) and cats (61.2%, 48.3%) across ComPath II and ComPath III, respectively. The percentage of resistance in E. coli was low (<10%) across both programmes in both dogs and cats except for trimethoprim-sulfamethoxazole (dogs ComPath III: 12.9%; cats ComPath II: 13.0%) and enrofloxacin (10.5%), marbofloxacin (11.4%), and doxycycline (98.8%) for dogs in ComPath III. Three (7.5%) of the 40 isolated S. aureus bacteria in total were MRSA and harboured mecA. The level of multidrug resistance (MDR) was generally low and ranged from 0.0% for feline coagulase-negative Staphylococcus spp. to 11.7% for canine Proteus spp., except for a peak of MDR observed in canine Klebsiella isolates from ComPath II (36.7%). Overall, antimicrobial resistance for most canine and feline UTI pathogens isolated during the ComPath II and ComPath III programmes was low (1-10%) to moderate (10-20%).

Pan-European monitoring of susceptibility to human-use antimicrobial agents in enteric bacteria isolated from healthy food-producing animals.

OBJECTIVES: To determine the antimicrobial susceptibility of Escherichia coli, Salmonella, Campylobacter and Enterococcus from cattle, pigs and chickens across the European Union (EU) using uniform methodology. METHODS: Intestinal samples (1624) were taken at slaughter across five EU countries. Bacteria were isolated in national laboratories, whilst MICs were determined in a central laboratory for key antimicrobials used in human medicine. Clinical resistance was based on CLSI breakpoints and decreased susceptibility based on European Food Safety Authority (EFSA)/EUCAST epidemiological cut-off values. RESULTS: Isolation rates were high for E. coli (n=1540), low for Salmonella (n=201) and intermediate for Campylobacter (n=940) and Enterococcus (n=786). For E. coli and Salmonella, clinical resistance to newer compounds (cefepime, cefotaxime and ciprofloxacin) was absent or low, but decreased susceptibility was apparent, particularly in chicken strains. Resistance to older compounds (except gentamicin) was variable and higher. Colistin resistance was absent for E. coli, but apparent for Salmonella. For Campylobacter jejuni, ciprofloxacin resistance was markedly prevalent for chickens, whereas clinical resistance and decreased susceptibility to erythromycin was absent or very low. For Campylobacter coli, resistance was notably higher. None of the Enterococcus faecium strains was resistant to linezolid, but some were resistant to ampicillin or vancomycin. Resistance to quinupristin/dalfopristin was frequent. CONCLUSIONS: Resistance patterns varied widely depending on bacterial species, antibiotics, hosts and region. Resistance varied among countries, particularly for older antimicrobials, but clinical resistance to newer antibiotics used to treat foodborne disease in humans was generally very low. In the absence of resistance to newer compounds in E. coli and Salmonella, the apparent decreased susceptibility should be monitored.

Genomic Diversity and Virulence Potential of ESBL- and AmpC-ß-Lactamase-Producing Escherichia coli Strains From Healthy Food Animals Across Europe.

The role of livestock animals as a putative source of ESBL/pAmpC E. coli for humans is a central issue of research. In a large-scale pan-European surveillance, 2,993 commensal Escherichia spp. isolates were recovered from randomly collected fecal samples of healthy cattle, pigs and chickens in various abattoirs. One-hundred Escherichia spp. isolates (0.5% from cattle, 1.3% pigs, 8.0% chickens) fulfilled the criteria for cefotaxime and ceftazidime non-wildtype (EUCAST). In silico screening of WGS data of 99 isolates (98 E. coli and 1 E. fergusonii) revealed bla SHV - 12 (32.3%), bla CTX - M - 1 (24.2%), and bla CMY - 2 (22.2%) as predominant ESBL/pAmpC types. Other types were bla SHV - 2 (1.0%), bla CTX - M - 2 / - 14 / - 15 (1.0/6.1/1.0%), and bla TEM - 52 (5.1%). Six isolates revealed AmpC-promoter mutations (position -42 (C > T) and one carried mcr-1. The majority (91.3%) of ESBL/pAmpC genes were located on plasmids. SHV-12 was mainly (50%) encoded on IncI1α plasmids (pST-3/-26/-95), followed by IncX3 (12.5%) and IncK2 (3.1%). The bla TEM - 52 genes were located on IncI1α-pST-36 (60%) and IncX1 plasmids (20%). The dominant plasmid lineage among CTX-M-1 isolates was IncI1α (pST-3/-295/-317) (87.5%), followed by IncN-pST-1 (8.3%). CMY-2 was mostly identified on IncI1α (pST-12/-2) (54.5%) and IncK2 (31.8%) plasmids. Several plasmids revealed high similarity to published plasmids from human and animal Enterobacteriaceae. The isolates were assigned to phylogroups A/C (34.7/7.1%), B1 (27.6%), B2 (3.1%), D/F (9.2/10.2%), E (5.1%), and to E. clades (3.0%). With 51 known and 2 novel MLST types, a wide variety of STs was found, including STs previously observed in human isolates (ST10/38/117/131/648). ESBL/AmpC types or STs were rarely correlated with the geographic origin of the isolates or animal species. Virulence gene typing identified extraintestinal pathogenic E. coli (ExPEC; 2.0%), avian pathogenic E. coli (APEC; 51.5%), and atypical enteropathogenic E. coli (EPEC; 6.1%). In conclusion, the high diversity of STs and phylogenetic groups provides hardly any hint for clonal spread of single lineages but hints toward the dissemination of cephalosporin resistance genes in livestock via distinct, globally successful plasmid lineages. Even though a number of isolates could not be assigned to a distinct pathotype, our finding of combined multidrug-resistance and virulence in this facultative pathogen should be considered an additional threat to public health.

Antimicrobial susceptibility monitoring of Mycoplasma hyopneumoniae isolated from seven European countries during 2015-2016.

Mycoplasma hyopneumoniae is the causative agent of porcine enzootic pneumonia, a chronic respiratory disease, causing significant economic losses. Results from the 2015-2016 MycoPath pan-European antimicrobial susceptibility monitoring survey of M. hyopneumoniae are presented. In total, 147 M. hyopneumoniae porcine isolates from Belgium, France, Germany, Great Britain, Hungary, Italy, and Spain were tested. One isolate per farm was retained from pigs that had not been recently treated with antimicrobial agents. The minimal inhibitory concentration (MIC) of 13 antimicrobial agents was determined in a central laboratory using a broth microdilution method, with Friis Medium, incubated at 35 ± 1 °C for 5-12 days. M. hyopneumoniae NCTC 10110 was used as Quality Control. MIC50/MIC90 (mg/L) values were: enrofloxacin 0.06/1; marbofloxacin 0.06/2; spiramycin 0.06/0.25; tulathromycin ≤0.001/0.004; gamithromycin 0.06/0.5; tylosin 0.016/0.06; tilmicosin 0.06/0.5; florfenicol 0.5/1; doxycycline 0.25/1; oxytetracycline 0.25/2; lincomycin 0.06/0.25; tiamulin 0.016/0.06 and valnemulin ≤0.001/0.004. Compared with the data from 2010 to 2012 MycoPath study (50 isolates), MIC50/90 results were similar and the majority were within ± two dilution steps, except for the MIC50 of oxytetracycline which is more than two dilution steps higher in the present study. Between-country comparisons show some differences in the MIC values for the fluoroquinolones, tulathromycin and tylosin, but the limited sample size per country precludes performing meaningful country comparisons for several countries. Standardized laboratory methods and interpretive criteria for MIC testing of veterinary mycoplasmas are clearly needed; there are currently no clinical breakpoints available to facilitate data interpretation and correlation of MICs with in vivo efficacy.

Antimicrobial susceptibility of nine udder pathogens recovered from bovine clinical mastitis milk in Europe 2015-2016: VetPath results.

VetPath is an ongoing pan-European antimicrobial susceptibility monitoring programme collecting pathogens from diseased cattle, pigs and poultry not recently treated with antibiotics. Non-duplicate isolates (n = 1244) were obtained from cows with acute clinical mastitis in eight countries during 2015-2016 for centrally antimicrobial susceptibility testing according CLSI standards. Among Escherichia coli (n = 225), resistance was high to ampicillin and tetracycline, moderate to kanamycin and low to amoxicillin/clavulanic acid and cefazolin. The MIC50/90 of danofloxacin, enrofloxacin and marbofloxacin were 0.03 and 0.06 µg/mL. For Klebsiella spp. (n = 70), similar results were noted, except for ampicillin and kanamycin. We detected 3.7 % (11/295) Enterobacteriaceae isolates carrying an ESBL/AmpC gene. Staphylococcus aureus (n = 247) and coagulase-negative staphylococci (CoNS; n = 189) isolates were susceptible to most antimicrobials tested except to penicillin (25.1 and 29.1 % resistance). Two S. aureus and thirteen CoNS isolates harboured mecA gene. Streptococcus uberis isolates (n = 208) were susceptible to ß-lactam antibiotics (87.1-94.7 % susceptibility), 23.9 % were resistant to erythromycin and 37.5 % to tetracycline. Resistance to pirlimycin was moderate. For Streptococcus dysgalactiae (n = 132) the latter figures were 10.6 and 43.2 %; pirlimycin resistance was low. MIC values for Streptococcus agalactiae, Trueperella pyogenes and Corynebacterium spp. were generally low. This current VetPath study shows that mastitis pathogens were susceptible to most antimicrobials with exceptions of staphylococci against penicillin and streptococci against erythromycin or tetracycline. For most antimicrobials, the percentage resistance and MIC50/90 values among the major pathogens were comparable to that of the preceeding VetPath surveys. This work highlights the high need to set additional clinical breakpoints for antimicrobials frequently used to treat mastitis.

A pan-European survey of antimicrobial susceptibility towards human-use antimicrobial drugs among zoonotic and commensal enteric bacteria isolated from healthy food-producing animals.

OBJECTIVES: The aim of the study was to study antimicrobial susceptibility in Escherichia coli, Salmonella, Campylobacter and Enterococcus recovered from chickens, pigs and cattle using uniform methodology. METHODS: Intestinal samples were taken at slaughter in five EU countries per host and bacteria isolated in national laboratories. MICs were determined in a central laboratory of key antimicrobials used in human medicine. Clinical resistance was based on CLSI breakpoints and decreased susceptibility on EFSA epidemiological cut-off values. RESULTS: Isolation rates from a total of 1500 samples were high for E. coli (n=1465), low for Salmonella (n=205) and intermediate for Campylobacter (n=785) and Enterococcus (n=718). Resistance prevalence varied among antibiotics, bacteria, hosts and countries. For E. coli and Salmonella, clinical resistance to newer compounds (cefepime, cefotaxime, ciprofloxacin) was absent or low, but a decreased susceptibility was apparent, particularly in chickens. Clinical resistance to older compounds (except colistin and gentamicin) was variable and higher. For Campylobacter jejuni from chickens, ciprofloxacin resistance was markedly higher than in isolates from cattle. Clinical resistance to erythromycin was absent for both hosts; decreased susceptibility very low. Similar trends were determined for Campylobacter coli, but C. jejuni was less resistant. None of the enterococcal strains was resistant to linezolid, but a few displayed resistance to ampicillin or vancomycin. Resistance prevalence to quinupristin/dalfopristin was clearly higher. CONCLUSIONS: Antimicrobial resistance among enteric organisms in food animals varied among countries, particularly for older antimicrobials, but clinical resistance to essential compounds used to treat disease in humans was generally zero or low. In the absence of clinical resistance to newer compounds in E. coli and Salmonella, the apparent decreased susceptibility should be monitored carefully.

New antimicrobial susceptibility data from monitoring of Mycoplasma bovis isolated in Europe.

Mycoplasma bovis is an important respiratory pathogen of cattle across Europe and is included in the MycoPath pan-European antimicrobial susceptibility monitoring programme. M. bovis strains (232) were isolated from cattle, not recently treated with antimicrobials, at diverse geographical locations in France, Great Britain, Hungary, Italy and Spain during 2014 to 2016. Only one isolate per farm and per outbreak was retained. For each isolate, the MICs of ten antimicrobials were determined in a central laboratory using a broth microdilution method with modified Eaton's medium and incubation at 35 °C ± 1 °C for 24 ± 6 h. MIC50/MIC90 (mg/L) values for the 232 strains were: danofloxacin 0.25/1; enrofloxacin 0.5/8; marbofloxacin 1/4; gamithromycin >64/>64; spiramycin 8/16; tilmicosin >64/>64; tulathromycin >64/>64; tylosin 64/>64; florfenicol 4/8; oxytetracycline 8/32. Minor between-country differences in the MIC90 values were observed for the fluoroquinolones, spiramycin and oxytetracycline, whilst the MIC values for the other compounds were similar. Spain and Italy had the higher MIC90 values for the fluoroquinolones. Compared with the 2010-2012 study (156 isolates) results are similar, with an overall MIC50 increase of at most one doubling dilution for enrofloxacin, spiramycin, tylosin, florfenicol and oxytetracycline. In contrast, the MIC90 value for oxytetracycline decreased from >64 to 32 mg/L. Standardized laboratory methods and interpretive criteria for MIC testing of veterinary mycoplasmas are clearly needed; there are currently no clinical breakpoints available to facilitate data interpretation and correlation of MICs with in vivo efficacy.

mcr-1-like detection in commensal Escherichia coli and Salmonella spp. from food-producing animals at slaughter in Europe.

We evaluate here the presence of the mcr-1-like and mcr-2 genes in Escherichia coli and Salmonella spp. isolated from healthy food-producing animals at slaughter between 2002 and 2014 in Europe. Isolates were retrieved from cattle, pig and chicken from 11 European countries of production. The susceptibility to colistin and antibiotics used in human medicine was determined by agar dilution. Colistin-resistant isolates were PCR-screened for mcr genes. mcr-positive isolates were typed by Pulsed-Field Gel Electrophoresis (PFGE) and Multi-Locus Sequence Typing. Among the 10,206 E. coli and 1774 Salmonella spp. isolated from cattle, pigs and chickens, 148 E. coli and 92 Salmonella spp. isolates were resistant to colistin. We found mcr-1-like gene in 68 (0.7%) E. coli and 2 (0.1%) Salmonella isolates whereas none of the isolates tested positive for mcr-2. MCR-1-like-positive E. coli were isolated from 2008 to 2014 in chicken (n=44, 1.2%) and pigs (n=24, 0.7%). The presence of mcr-1-like varied from 0 to 4.0% depending on the year and the animal species. mcr-1-like-positive isolates came from animals originating from Germany (n=38), Spain (n=23), The Netherlands (n=5), and France (n=4). They were distributed in 63 different PFGE types and 37 different STs, with ST10 being the most prevalent. The two mcr-1-like-positive Salmonella spp. were isolated from France and Germany from a pig and a chicken, respectively. mcr-1-like gene is present in food-producing animals at slaughter in European countries with the highest occurrence in chickens. The high clonal diversity of E. coli underlines the evidence for horizontal transfer of mcr-1-like genes.

Monitoring of antimicrobial susceptibility of udder pathogens recovered from cases of clinical mastitis in dairy cows across Europe: VetPath results.

VetPath is an ongoing pan-European antimicrobial susceptibility monitoring programme collecting pathogens from diseased cattle, pigs and poultry not recently treated with antibiotics. Non-duplicate milk samples were collected from cows with acute clinical mastitis in nine countries and 934 isolates were obtained during 2009-2012 for subsequent antimicrobial susceptibility testing in a central laboratory. CLSI broth microdilution methodology was used, and where available, MICs were interpreted using CLSI approved veterinary-specific (ceftiofur) otherwise human clinical breakpoints. Among Escherichia coli (n=207) and Klebsiella spp., (n=87), resistance was moderate to tetracycline and high to cephapirin (E. coli only) whereas resistance to other ß-lactam antibiotics was very low (ceftiofur) to low (amoxicillin/clavulanic acid, cephalexin, cephalonium). The MIC90 of enrofloxacin and marbofloxacin was 0.03 and 0.06µg/mL respectively (E. coli) with 0.5% strains displaying higher MICs. Staphylococcus aureus (n=192) and coagulase-negative staphylococci (CNS; n=165) strains were susceptible to most antibiotics tested except to penicillin (25.0 and 29.1% resistance), respectively. Three S. aureus and seven CNS strains were oxacillin-resistant and harboured mecA. Streptococcus uberis strains (n=188) were susceptible to the ß-lactam antibiotics although 35.6% were penicillin intermediately susceptible, and 20.2% were resistant to erythromycin, 36.7% to tetracycline. For Streptococcus dysgalactiae (n=95) the latter figures were 13.7 and 56.8%, respectively. For most antibiotics, the percentage resistance among E. coli, S. aureus and S. uberis was comparable to that of the VetPath 2002-2006 survey. This current, expanded VetPath study shows that mastitis pathogens were susceptible to most antibiotics with exceptions of staphylococci tested against penicillin and streptococci against erythromycin or tetracycline. This work highlights the high need to set additional clinical breakpoints for antibiotics frequently used to treat mastitis.

Characterization of quinolone resistance mechanisms in Enterobacteriaceae isolated from companion animals in Europe (ComPath II study).

ComPath is an ongoing European programme dedicated to monitor antibiotic susceptibility of bacterial pathogens from diseased dogs and cats. The objective was to characterize determinants associated with quinolone resistance among 160 enrofloxacin non-wild type strains (100 Escherichia coli, 45 Proteus mirabilis, 14 Klebsiella pneumoniae, 1 Enterobacter cloacae) selected among 843 non-duplicate Enterobacteriaceae isolates collected in 12 European countries (2013-2014). These strains with non-wild type MICs of ≥0.25 mg/L, for P. mirabilis ≥0.5 mg/L, were screened for PMQR determinants (qnr, oqxAB, qepA and aac(6')-Ib-cr), and for QRDR mutations in gyrA, gyrB, parC and parE genes. Among them, 20% (32/160) carried at least one PMQR (18/32 qnrB, qnrS or qnrD, 10/32 aac(6')-Ib-cr and 13/32 oqxAB), and 80% (128/160) no PMQR. qnrB was detected in 3 E. coli, 2 K. pneumoniae and 1 E. cloacae strains; qnrS in 6 E. coli and 1 P. mirabilis and aac(6')-Ib-cr in 4 E. coli, 5 K. pneumoniae and 1 E. cloacae strains. All qnrD1 were detected in P. mirabilis. oqxAB was detected in 12/14 K. pneumoniae and 1 E. cloacae. No qepA genes were detected. From the 32 PMQR-positive strains, 10 showed enrofloxacin MICs ≤2 mg/L and 22 MICs ≥8 mg/L, the latter carrying 1-4 mutations in QRDR. For the 128 non-PMQR strains, 37 showed enrofloxacin MICs ≤2 mg/L with 0-2 QRDR mutations, and 91 MICs ≥4 mg/L carrying 1-4 QRDR mutations. In conclusion, qnr was the major PMQR and qnrD only detected in Proteeae. Mutations in QRDR play a markedly greater role in mediating fluoroquinolone resistance than PMQR.

Antimicrobial susceptibility of enterococci recovered from healthy cattle, pigs and chickens in nine EU countries (EASSA Study) to critically important antibiotics.

The European Antimicrobial Susceptibility Surveillance in Animals (EASSA) program collects zoonotic and commensal bacteria from food-producing animals at slaughter and tracks their susceptibility to medically important antibiotics. Results of commensal enterococci species (2013-2014) are presented here. Intestinal content from cattle, pigs and chickens were randomly sampled (5-6 countries/host; ≥4 abattoirs/country; 1 sample/animal/farm) for isolation of enterococci, MICs of 9 antibiotics were assessed by CLSI agar dilution in a central laboratory. Clinical breakpoints (CLSI) and epidemiological cut-off values (EUCAST) were applied for data interpretation. In total 960 Enterococcus faecium and 779 Enterococcus faecalis strains were recovered. Seven porcine E. faecium/faecalis strains of Spanish origin were resistant to linezolid. One avian E. faecalis and one porcine E. faecium strain were non-wild type (MICs 8 mg/L) to daptomycin. Clinical vancomycin resistance was absent; 2 poultry E. faecium and 1 bovine E. faecalis strains were non-wild type, all with MICs of 8 mg/L. None of the strains tested were clinically resistant to tigecycline. Little clinical resistance to ampicillin or gentamicin was observed. Clinical resistance of E. faecium to quinupristin/dalfopristin was slightly higher (2.2-12.0%) but 61.9-83.2% of the strains were classified as non-wild type. Very high percentages resistance to tetracycline (67.4-78.3%) and to erythromycin (27.1-57.0%) were noted for both E. faecium and E. faecalis in pigs and chickens compared to cattle (5.2-30.4 and 9.0-10.4%, respectively). Similar non-wild type results were observed for E. hirae (n = 557), E. durans (n = 218) and E. casseliflavus (n = 55) including percentage non-wild type for daptomycin, linezolid, tigecycline being absent and for vancomycin low. For these species percentage non-wild type to erythromycin was lower as compared to E. faecalis/faecium. This pan-EU survey shows high variability in antibiotic susceptibility of commensal enterococci from healthy food animals. Clinical resistance to critically important antibiotics for human medicine was absent or low, except for erythromycin.

Re-analysis of the risks attributed to ciprofloxacin-resistant Campylobacter jejuni infections.

Recent publications were compared and analysed in addition to novel surveillance data to assess the hypothesis that fluoroquinolone-resistant Campylobacter infections are causing more severe disease than susceptible infections. The available data did not support this hypothesis. There was no significant difference in duration of disease between susceptible and resistant infections. However, both for resistant and susceptible infections, disease symptoms were prolonged by on average 1-2 days for Campylobacter cases acquired during foreign travel. Nevertheless, the likelihood and duration of hospitalisation were not increased for cases related to foreign travel. These observations were confirmed by a new analysis of almost 11,000 cases. We conclude that fluoroquinolone-resistant Campylobacter infections are not more severe than susceptible infections.

Detection of a Novel qnrB19-Carrying Plasmid Variant Mediating Decreased Fluoroquinolone Susceptibility in Salmonella enterica Serovar Hadar.

Thirty Salmonella enterica subsp. enterica serovar Hadar isolates of avian origin collected between 2007 and 2010 from chicken carcasses in five geographically spread abattoirs in Germany were investigated for plasmid-mediated quinolone resistance determinants. Four isolates were identified by PCR analysis and hybridization experiments to carry qnrB genes. The isolates were indistinguishable by their XbaI macrorestriction patterns and did not exhibit a mutation in the quinolone resistance-determining regions of the DNA gyrase and topoisomerase IV genes. The qnrB genes were found to be located on small plasmids of ∼2.6 kb, which mediated decreased susceptibility only to quinolones. The plasmids were assigned to the same type, pHAD28, and transformation studies into an Escherichia coli recipient strain confirmed their transferability. Sequence analysis of the complete plasmid pHAD28 revealed the presence of a qnrB19 gene. The gene was found on a novel variant of qnrB19-harboring plasmids with high similarity to plasmids pPAB19-3 from E. coli and pPAB19-4 from Salmonella sp. M9397. A presumptive recombination side was detected, suggesting that interplasmid recombination events might have played a role in the development of this plasmid variant.

Antimicrobial Susceptibility Monitoring of Bacterial Pathogens Isolated from Urinary Tract Infections in Dogs and Cats Across Europe: ComPath Results.

ComPath is a pan-European antimicrobial surveillance program collecting bacterial pathogens from dogs and cats not recently exposed to antimicrobials. We present minimum inhibitory concentration data obtained using Clinical and Laboratory Standards Institute methodology for 616 urinary tract infection (UTI) isolates collected between 2008 and 2010. In both dogs and cats, the most common pathogen was Escherichia coli (59.8% and 46.7%, respectively). Antimicrobial activity against E. coli in dogs and cats was similar with fluoroquinolone and trimethoprim/sulfamethoxazole susceptibility >90%. Ampicillin susceptibility was ∼80%. Staphylococcus intermedius Group isolates from dogs (67/437, 15.3%) had high antimicrobial susceptibility (>90%) toward beta-lactams, fluoroquinolones, and trimethoprim/sulfamethoxazole. Four canine isolates (6%) were oxacillin resistant, and harbored mecA. Proteus mirabilis from dogs (48/437, 11.0%) had high antimicrobial susceptibility (∼90%) to amoxicillin/clavulanic acid, enrofloxacin, and marbofloxacin and slightly lower susceptibility (∼80-85%) to ampicillin and orbifloxacin. Streptococcus canis isolates (35/437, 8.0%) from dogs were all susceptible to ampicillin and amoxicillin/clavulanic acid and >90% susceptible to marbofloxacin. Although resistance was not observed, high intermediate susceptibility was seen for both enrofloxacin (28.6%) and orbifloxacin (85.7%). Overall, antimicrobial in vitro activity appears to be high in UTI pathogens from dogs and cats with low multidrug resistance, although a lack of specific dog and cat breakpoints for important antimicrobials such as cefovecin, cephalexin, and ibafloxacin prevents analysis of susceptibility for these agents.

Antimicrobial susceptibility monitoring of Mycoplasma hyopneumoniae and Mycoplasma bovis isolated in Europe.

Mycoplasma hyopneumoniae in pigs and Mycoplasma bovis in cattle are major pathogens affecting livestock across Europe and are the focus of the MycoPath pan-European antimicrobial susceptibility monitoring programme. Fifty M. hyopneumoniae isolates from Belgium, Spain and the United Kingdom (UK), and 156 M. bovis isolates from France, Hungary, Spain and the UK that met specific criteria were tested for antimicrobial susceptibility in a central laboratory by using a microbroth dilution method. Specific isolate criteria included recovery from animals not recently treated with antimicrobials, isolates from different locations within each country and retaining only one isolate per farm. MIC50/MIC90 values were 0.031/0.5, 0.031/0.5, 0.062/0.25, ≤0.001/0.004, 0.031/0.125, 0.25/0.5 and 0.062/0.25mg/L for enrofloxacin, marbofloxacin, spiramycin, tulathromycin, tylosin, florfenicol and oxytetracycline respectively against M. hyopneumoniae and 0.25/4, 1/4, 4/16, >64/ >64, 32/ >64, 2/4 and 4/64mg/L, respectively against M. bovis. MIC50/MIC90 values for tiamulin and valnemulin against M. hyopneumoniae were 0.016/0.062 and ≤0.001/ ≤0.001mg/L respectively. The MIC50/MIC90 values of danofloxacin and gamithromycin for M. bovis were 0.25/1 and >64/ >64mg/L respectively. The highest MIC90 values for M. hyopneumoniae were found in the UK at 1.0mg/L for enrofloxacin, marbofloxacin and florfenicol. In contrast, for M. bovis the lowest MIC90 value was 1.0mg/L, but ranged to >64mg/L. Specific laboratory standards and clinical breakpoints for veterinary Mycoplasma species are required as no independently validated clinical breakpoints are specified for veterinary Mycoplasma species, which makes data interpretation and correlation to in vivo efficacy difficult.