Trends in antimicrobial resistance among Escherichia coli from defined infections in humans and animals.

OBJECTIVES: To characterize and compare resistance trends in clinical Escherichia coli isolates from humans, food-producing animals (poultry, cattle and swine) and pets (dogs and cats). METHODS: Antibiogram results collected between January 2014 and December 2017 by MedQual [the French surveillance network for antimicrobial resistance (AMR) in bacteria isolated from the community] and RESAPATH (the French surveillance network for AMR in bacteria from diseased animals) were analysed, focusing on resistance to antibiotics of common interest to human and veterinary medicine. Resistance dynamics were investigated using generalized additive models. RESULTS: In total, 743 637 antibiograms from humans, 48 170 from food-producing animals and 7750 from pets were analysed. For each antibiotic investigated, the resistance proportions of isolates collected from humans were of the same order of magnitude as those from food-producing animals or pets. However, resistance trends in humans differed from those observed in pets and food-producing animals over the period studied. For example, resistance to third-generation cephalosporins and fluoroquinolones was almost always below 10% for both humans and animals. However, in contrast to the notable decreases in resistance observed in both food-producing animals and pets, resistance in humans decreased only slightly. CONCLUSIONS: Despite several potential biases in the data, the resistance trends remain meaningful. The strength of the parallel is based on similar data collection in humans and animals and on a similar statistical methodology. Resistance dynamics seemed specific to each species, reflecting different antibiotic-use practices. These results advocate applying the efforts already being made to reduce antibiotic use to all sectors and all species, both in human and veterinary medicine.

Antimicrobial resistance in bacteria isolated from mastitis in dairy cattle in France, 2006-2016.

In dairy cattle, mastitis is the most frequent bacterial disease, and the routine use of antibiotics for treatment and prevention can drive antimicrobial resistance (AMR). The aim of our study was to estimate the levels of AMR of the 3 main bacteria isolated from dairy cattle with mastitis in France (Streptococcus uberis, Escherichia coli, and coagulase-positive staphylococci) and to investigate their changes over time. Data collected between 2006 and 2016 by the French surveillance network for AMR in pathogenic bacteria of animal origin (called RESAPATH) were analyzed. The proportions of mono- and multidrug resistance were calculated and the trends were investigated using nonlinear analyses applied to time series. Over the whole period, the lowest proportions of resistance in S. uberis isolates were observed for oxacillin (2.2%) and gentamicin (2.4%) and most resistance levels were below 20%. The trends in resistance showed some significant variation, mainly for S. uberis, but without a common pattern across the various antibiotics examined. For only 2 combinations of bacteria-antibiotic the trend in resistance showed a continuous increase from 2006 to 2016: tetracycline resistance in S. uberis isolates and third-generation cephalosporin resistance in E. coli isolates. In E. coli, the highest proportions of resistance were observed for amoxicillin (28.1%) and tetracycline (23.1%). Resistance to third-generation cephalosporins in E. coli from dairy cattle was almost nil in 2006, but reached 2.4% in December 2016. This increase is particularly concerning because these antibiotics constitute one of the latest therapeutic alternatives to fight severe infectious diseases in humans. Except for penicillin (33.9%), the proportions of resistance in coagulase-positive staphylococci were below 11% during the whole study period. Multidrug resistance (isolates with acquired resistance to at least one antibiotic in 3 or more antibiotic classes) ranged from 2.4% for coagulase-positive staphylococci to 9.9% for S. uberis. These findings can serve as guidelines for practitioners in the choice of the most appropriate antibiotic according to the prevailing epidemiological context. Ultimately, our results contribute to risk assessment of AMR and provide a baseline for setting up and evaluating control measures and designing strategies to limit AMR.

Impact of heat and cold waves on female cattle mortality beyond the effect of extreme temperatures.

Although heat and cold temperatures are known to have an impact on cattle mortality, no study has evidenced and quantified the influence of the prolonged exposure to extreme temperatures beyond the single effect of daily temperatures. We defined a heat (or cold) wave by a continuous variable indicating the number of successive days with temperatures above (or below) a given threshold. For heat wave, the threshold was set to the 95th or 99th percentile of the mean daily temperature distribution and for cold wave to the 1st or 5th percentile. We collected female cattle mortality data by type of production and age classes between 2001 and 2015 for 100 iso-hygro-thermal areas in France. We used time-series analyses to estimate the area-specific heat wave- and cold wave-mortality relationships. Then, we applied meta-analyses to pool area-specific effects at the country level for each definition of heat and cold wave. For each type of production and age classes, our models predicted symmetrical relationships between temperature and mortality, with a temperature range of minimum mortality located approximately between 15 and 20 °CTHI in most categories. Outside that range, relative risks between 1.3 and 2.5 were estimated for extreme cold temperatures and relative risks between 1.1 and 1.5 were estimated for extreme hot temperatures depending on age categories and production type. Our results indicated that a prolonged exposure to high (or low) temperatures caused a significant increase on mortality (up to 40% during heat waves and 23% for cold waves, depending on type of production and age classes), in addition to the effect of extreme temperature alone. This additional mortality risk increased along with the duration and intensity of the exposure. Our results suggest that not discriminating the effect of the prolonged exposure to extreme temperature, may overestimate the effect of temperature alone on mortality.

Prevalence of carriage of Shiga toxin-producing Escherichia coli serotypes O157:H7, O26:H11, O103:H2, O111:H8, and O145:H28 among slaughtered adult cattle in France.

The main pathogenic enterohemorrhagic Escherichia coli (EHEC) strains are defined as Shiga toxin (Stx)-producing E. coli (STEC) belonging to one of the following serotypes: O157:H7, O26:H11, O103:H2, O111:H8, and O145:H28. Each of these five serotypes is known to be associated with a specific subtype of the intimin-encoding gene (eae). The objective of this study was to evaluate the prevalence of bovine carriers of these "top five" STEC in the four adult cattle categories slaughtered in France. Fecal samples were collected from 1,318 cattle, including 291 young dairy bulls, 296 young beef bulls, 337 dairy cows, and 394 beef cows. A total of 96 E. coli isolates, including 33 top five STEC and 63 atypical enteropathogenic E. coli (aEPEC) isolates, with the same genetic characteristics as the top five STEC strains except that they lacked an stx gene, were recovered from these samples.O157:H7 was the most frequently isolated STEC serotype. The prevalence of top five STEC (all serotypes included) was 4.5% in young dairy bulls, 2.4% in young beef bulls, 1.8% in dairy cows, and 1.0% in beef cows. It was significantly higher in young dairy bulls (P<0.05) than in the other 3 categories. The basis for these differences between categories remains to be elucidated. Moreover,simultaneous carriage of STEC O26:H11 and STEC O103:H2 was detected in one young dairy bull. Lastly, the prevalence of bovine carriers of the top five STEC, evaluated through a weighted arithmetic mean of the prevalence by categories, was estimated to 1.8% in slaughtered adult cattle in France.

Quantifying the influence of ambient temperature on dairy and beef cattle mortality in France from a time-series analysis.

In the context of climate change, the frequency and severity of extreme weather events are expected to increase in temperate regions, and potentially have a severe impact on farmed cattle through production losses or deaths. In this study, we used distributed lag non-linear models to describe and quantify the relationship between a temperature-humidity index (THI) and cattle mortality in 12 areas in France. THI incorporates the effects of both temperature and relative humidity and was already used to quantify the degree of heat stress on dairy cattle because it does reflect physical stress deriving from extreme conditions better than air temperature alone. Relationships between daily THI and mortality were modeled separately for dairy and beef cattle during the 2003-2006 period. Our general approach was to first determine the shape of the THI-mortality relationship in each area by modeling THI with natural cubic splines. We then modeled each relationship assuming a three-piecewise linear function, to estimate the critical cold and heat THI thresholds, for each area, delimiting the thermoneutral zone (i.e. where the risk of death is at its minimum), and the cold and heat effects below and above these thresholds, respectively. Area-specific estimates of the cold or heat effects were then combined in a hierarchical Bayesian model to compute the pooled effects of THI increase or decrease on dairy and beef cattle mortality. A U-shaped relationship, indicating a mortality increase below the cold threshold and above the heat threshold was found in most of the study areas for dairy and beef cattle. The pooled estimate of the mortality risk associated with a 1°C decrease in THI below the cold threshold was 5.0% for dairy cattle [95% posterior interval: 4.4, 5.5] and 4.4% for beef cattle [2.0, 6.5]. The pooled mortality risk associated with a 1°C increase above the hot threshold was estimated to be 5.6% [5.0, 6.2] for dairy and 4.6% [0.9, 8.7] for beef cattle. Knowing the thermoneutral zone and temperature effects outside this zone is of primary interest for farmers because it can help determine when to implement appropriate preventive and mitigation measures.

Assessment of Adhesins as an Indicator of Pathovar-Associated Virulence Factors in Bovine Escherichia coli.

The CS31A, F17, and F5 adhesins are usually targeted by serology-based methods to detect pathogenic Escherichia coli associated with calf enteritis. However, the virulence traits of the selected isolates are still poorly described. Here, from a set of 349 diarrheagenic E. coli isolates from cattle, we demonstrated a 70.8% concordance rate (Cohen's kappa, 0.599) between serology- and PCR-based approaches for the detection of adhesins under field conditions. A 79% to 82.4% correspondence between the two methods was found for fimbrial adhesins, whereas major discrepancies (33%) were observed for CS31A-type antigens. Various F17A variants were found, such as F17Ac (20K) (50%), F17Aa (FY) (18.9%), F17Ab (8.1%), and F17Ad (111K) (5.4%), including a high proportion (17.6%) of new F17A internal combinations (F17Aab, F17Aac, and F17Abc) or untypeable variants. In addition, the highest proportion of pathovar-associated virulence factor (VF) genes was observed among E. coli isolates that produced F5/F41 adhesins. A specific link between the heat-stable toxins related to the enterotoxigenic E. coli (ETEC) pathovar and adhesins was identified. STa was significantly linked to F5/F41 and EAST1 to CS31A adhesins (P < 0.001), respectively, whereas NTEC was associated with F17 adhesin (P = 0.001). Clustering between phylogroups according to the adhesin types was also observed. Also, few Shiga toxin-producing E. coli (STEC) or enteropathogenic E. coli (EPEC) pathovars were identified. Finally, no statistically significant difference was observed in the occurrence of extended-spectrum beta lactamase (ESBL) production according to the adhesins expressed by the isolates (P = 0.09). Altogether, this study gives new insights into the relationship between adhesins, VF, and antimicrobial resistance in calf enteritis and supports the need for further standardization of methodologies for such approaches.

Intimin gene (eae) subtype-based real-time PCR strategy for specific detection of Shiga toxin-producing Escherichia coli serotypes O157:H7, O26:H11, O103:H2, O111:H8, and O145:H28 in cattle feces.

Shiga toxin-producing Escherichia coli (STEC) strains belonging to serotypes O157:H7, O26:H11, O103:H2, O111:H8, and O145:H28 are known to be associated with particular subtypes of the intimin gene (eae), namely, γ1, ß1, ε, θ, and γ1, respectively. This study aimed at evaluating the usefulness of their detection for the specific detection of these five main pathogenic STEC serotypes in cattle feces. Using real-time PCR assays, 58.7% of 150 fecal samples were found positive for at least one of the four targeted eae subtypes. The simultaneous presence of stx, eae, and one of the five O group markers was found in 58.0% of the samples, and the five targeted stx plus eae plus O genetic combinations were detected 143 times. However, taking into consideration the association between eae subtypes and O group markers, the resulting stx plus eae subtype plus O combinations were detected only 46 times. The 46 isolation assays performed allowed recovery of 22 E. coli strains belonging to one of the five targeted STEC serogroups. In contrast, only 2 of 39 isolation assays performed on samples that were positive for stx, eae and an O group marker, but that were negative for the corresponding eae subtype, were successful. Characterization of the 24 E. coli isolates showed that 6 were STEC, including 1 O157:H7, 3 O26:H11, and 2 O145:H28. The remaining 18 strains corresponded to atypical enteropathogenic E. coli (aEPEC). Finally, the more discriminating eae subtype-based PCR strategy described here may be helpful for the specific screening of the five major STEC in cattle feces.

Defining syndromes using cattle meat inspection data for syndromic surveillance purposes: a statistical approach with the 2005-2010 data from ten French slaughterhouses.

BACKGROUND: The slaughterhouse is a central processing point for food animals and thus a source of both demographic data (age, breed, sex) and health-related data (reason for condemnation and condemned portions) that are not available through other sources. Using these data for syndromic surveillance is therefore tempting. However many possible reasons for condemnation and condemned portions exist, making the definition of relevant syndromes challenging.The objective of this study was to determine a typology of cattle with at least one portion of the carcass condemned in order to define syndromes. Multiple factor analysis (MFA) in combination with clustering methods was performed using both health-related data and demographic data. RESULTS: Analyses were performed on 381,186 cattle with at least one portion of the carcass condemned among the 1,937,917 cattle slaughtered in ten French abattoirs. Results of the MFA and clustering methods led to 12 clusters considered as stable according to year of slaughter and slaughterhouse. One cluster was specific to a disease of public health importance (cysticercosis). Two clusters were linked to the slaughtering process (fecal contamination of heart or lungs and deterioration lesions). Two clusters respectively characterized by chronic liver lesions and chronic peritonitis could be linked to diseases of economic importance to farmers. Three clusters could be linked respectively to reticulo-pericarditis, fatty liver syndrome and farmer's lung syndrome, which are related to both diseases of economic importance to farmers and herd management issues. Three clusters respectively characterized by arthritis, myopathy and Dark Firm Dry (DFD) meat could notably be linked to animal welfare issues. Finally, one cluster, characterized by bronchopneumonia, could be linked to both animal health and herd management issues. CONCLUSION: The statistical approach of combining multiple factor analysis with cluster analysis showed its relevance for the detection of syndromes using available large and complex slaughterhouse data. The advantages of this statistical approach are to i) define groups of reasons for condemnation based on meat inspection data, ii) help grouping reasons for condemnation among a list of various possible reasons for condemnation for which a consensus among experts could be difficult to reach, iii) assign each animal to a single syndrome which allows the detection of changes in trends of syndromes to detect unusual patterns in known diseases and emergence of new diseases.

Phylogenetic grouping and virulence potential of extended-spectrum-ß-lactamase-producing Escherichia coli strains in cattle.

In line with recent reports of extended-spectrum beta-lactamases (ESBLs) in Escherichia coli isolates of highly virulent serotypes, such as O104:H4, we investigated the distribution of phylogroups (A, B1, B2, D) and virulence factor (VF)-encoding genes in 204 ESBL-producing E. coli isolates from diarrheic cattle. ESBL genes, VFs, and phylogroups were identified by PCR and a commercial DNA array (Alere, France). ESBL genes belonged mostly to the CTX-M-1 (65.7%) and CTX-M-9 (27.0%) groups, whereas those of the CTX-M-2 and TEM groups were much less represented (3.9% and 3.4%, respectively). One ESBL isolate was stx(1) and eae positive and belonged to a major enterohemorrhagic E. coli (EHEC) serotype (O111:H8). Two other isolates were eae positive but stx negative; one of these had serotype O26:H11. ESBL isolates belonged mainly to phylogroup A (55.4%) and, to lesser extents, to phylogroups D (25.5%) and B1 (15.6%), whereas B2 strains were quasi-absent (1/204). The number of VFs was significantly higher in phylogroup B1 than in phylogroups A (P = 0.04) and D (P = 0.02). Almost all of the VFs detected were found in CTX-M-1 isolates, whereas only 64.3% and 33.3% of them were found in CTX-M-9 and CTX-M-2 isolates, respectively. These results indicated that the widespread dissemination of the bla(CTX-M) genes within the E. coli population from cattle still spared the subpopulation of EHEC/Shiga-toxigenic E. coli (STEC) isolates. In contrast to other reports on non-ESBL-producing isolates from domestic animals, B1 was not the main phylogroup identified. However, B1 was found to be the most virulent phylogroup, suggesting host-specific distribution of virulence determinants among phylogenetic groups.

Individual factors associated with L- and H-type Bovine Spongiform encephalopathy in France.

BACKGROUND: Cattle with L-type (L-BSE) and H-type (H-BSE) atypical Bovine Spongiform encephalopathy (BSE) were identified in 2003 in Italy and France respectively before being identified in other countries worldwide. As of December 2011, around 60 atypical BSE cases have currently been reported in 13 countries, with over one third in France. While the epidemiology of classical BSE (C-BSE) has been widely described, atypical BSEs are still poorly documented, but appear to differ from C-BSE. We analysed the epidemiological characteristics of the 12 cases of L-BSE and 11 cases of H-BSE detected in France from January 2001 to late 2009 and looked for individual risk factors. As L-BSE cases did not appear to be homogeneously distributed throughout the country, two complementary methods were used: spatial analysis and regression modelling. L-BSE and H-BSE were studied separately as both the biochemical properties of their pathological prion protein and their features differ in animal models. RESULTS: The median age at detection for L-BSE and H-BSE cases was 12.4 (range 8.4-18.7) and 12.5 (8.3-18.2) years respectively, with no significant difference between the two distributions. However, this median age differed significantly from that of classical BSE (7.0 (range 3.5-15.4) years). A significant geographical cluster was detected for L-BSE. Among animals over eight years of age, we showed that the risk of being detected as a L-BSE case increased with age at death. This was not the case for H-BSE. CONCLUSION: To the best of our knowledge this is the first study to describe the epidemiology of the two types of atypical BSE. The geographical cluster detected for L-BSE could be partly due to the age structure of the background-tested bovine population. Our regression analyses, which adjusted for the effect of age and birth cohort showed an age effect for L-BSE and the descriptive analysis showed a particular age structure in the area where the cluster was detected. No birth cohort effect was evident. The relatively small number of cases of atypical BSE and the few individual data available for the tested population limited our analysis to the investigation of age and cohort effect only. We conclude that it is essential to maintain BSE surveillance to further elucidate our findings.

Comparative assessment of two commonly used commercial ELISA tests for the serological diagnosis of contagious agalactia of small ruminants caused by Mycoplasma agalactiae.

BACKGROUND: Contagious agalactia (CA) of sheep and goats caused by Mycoplasma agalactiae is a widely occurring economically important disease that is difficult to control. The ELISA is commonly used for the serological detection of CA but it has some limitations and the performance of the available tests have not been properly evaluated.Two commercial ELISA kits are widely used, one involving a fusion protein as target antigen and the other a total antigen. The objectives were to compare these tests by evaluating:i. Their diagnostic sensitivity and specificity, the relevance of the recommended cut-off points, the correlation between the two tests, and, the correlation between serology data and the milk shedding of M. agalatiae;ii. The influence of extrinsic factors such as the targeted animal species, geographical origin of the samples, intra-specific variability of M. agalactiae and concurrent mycoplasma infections.A sample of 5900 animals from 211 farms with continuous CA monitoring for 20 years and no prior vaccination history was used. The infection status was known from prior bacteriological, epidemiological and serological monitoring with a complementary immunoblotting test. RESULTS: The average diagnostic sensitivity was 56% [51.8-59.8] for the fusion protein ELISA and 84% [81.3-87.2] for the total antigen ELISA, with noteworthy flock-related variations. The average diagnostic specificity for the fusion protein ELISA was 100% [99.9-100], and for the total antigen ELISA differed significantly between goats and sheep: 99.3% [97.4-99.9] and 95.7% [93.8-97.2] respectively.Experimental inoculations with different M. agalactiae strains revealed that the ELISA kits poorly detected the antibody response to certain strains. Furthermore, test performances varied according to the host species or geographical origin of the samples.Finally, the correlation between milk shedding of M. agalactiae and the presence of detectable antibodies in the blood was poor. CONCLUSIONS: These serological tests are not interchangeable. The choice of a test will depend on the objectives (early detection of infection or disease control program), on the prevalence of infection and the control protocol used. Given the variety of factors that may influence performance, a preliminary assessment of the test in a given situation is recommended prior to widespread use.

Quantitative assessments of indoor air pollution and respiratory health in a population-based sample of French dwellings.

BACKGROUND: Various volatile organic compounds (VOCs) have been related to respiratory health effects, but have generally been assessed individually without taking into account the fact that such pollutants are highly correlated to one other. AIMS: We investigated the effects of exposure to various VOC, and considered their combined effect on adult asthma and rhinitis. METHOD: A national cross-sectional representative survey conducted by the Indoor Air Quality Observatory objectively assessed 20 VOCs in 490 main dwellings in France. A standardized questionnaire determined the prevalence of asthma and rhinitis among 1012 inhabitants of the dwellings (≥ 15 years). Marginal models for binary outcome were used to relate VOCs exposure to asthma and rhinitis, controlling for potential confounders. A global score representing the number of VOCs in each dwelling with an elevated concentration (using the 3(rd) quartile value of the distribution as a threshold value) was then derived as a measure of the combined effect of VOCs. Specific scores were built using a similar approach, grouping VOCs by family. RESULTS: Asthma (8.6%) was significantly associated with N-undecane and 1,2,4-trimethylbenzene and rhinitis (38.3%) with ethylbenzene, trichloroethylene, m/p- and o-xylene. The global VOC score was associated with a significant risk of asthma and rhinitis (odds ratio (OR) of 1.40 and 1.22, respectively, for 5 additional VOCs with high exposure level). Both specific scores for aromatic hydrocarbons and aliphatic hydrocarbons were associated with a significantly risk of asthma (OR=1.12; 95% confidence interval (CI): 1.01-1.24 and OR=1.41; 95% CI=1.03-1.93, respectively). The specific VOC score for halogenated hydrocarbons was associated with a significant risk of rhinitis (OR=1.28; 95% CI: 1.07-1.54). CONCLUSION: We have shown that high concentrations of VOCs in homes were associated with an increasing prevalence of asthma and rhinitis in adults.

Drivers of ESBL-producing Escherichia coli dynamics in calf fattening farms: A modelling study.

The contribution of bacteria in livestock to the global burden of antimicrobial resistance raises concerns worldwide. However, the dynamics of selection and diffusion of antimicrobial resistance in farm animals are not fully understood. Here, we used veal calf fattening farms as a model system, as they are a known reservoir of Extended Spectrum ß-Lactamase-producing Escherichia coli (ESBL-EC). Longitudinal data of ESBL-EC carriage and antimicrobial use (AMU) were collected from three veal calf farms during the entire fattening process. We developed 18 agent-based mechanistic models to assess different hypotheses regarding the main drivers of ESBL-EC dynamics in calves. The models were independently fitted to the longitudinal data using Markov Chain Monte Carlo and the best model was selected. Within-farm transmission between individuals and sporadic events of contamination were found to drive ESBL-EC dynamics on farms. In the absence of AMU, the median carriage duration of ESBL-EC was estimated to be 19.6 days (95% credible interval: [12.7; 33.3]). In the best model, AMU was found to influence ESBL-EC dynamics, by affecting ESBL-EC clearance rather than acquisition. This effect of AMU was estimated to decrease gradually after the end of exposure and to disappear after 62.5 days [50.0; 76.9]. Moreover, using a simulation study, we quantified the efficacy of ESBL-EC mitigation strategies. Decreasing ESBL-EC prevalence by 50% on arrival at the fattening farm reduced prevalence at slaughter age by 33.3%. Completely eliminating the use of selective antibiotics on arrival had a strong effect on average ESBL-EC prevalence (relative reduction of 77.0%), but the effect was mild if this use was only decreased by 50% compared to baseline (relative reduction of 3.3%).

Co-resistance to Amoxicillin and Tetracycline as an Indicator of Multidrug Resistance in Escherichia coli Isolates From Animals.

OBJECTIVES: To examine the relevance of co-resistance to amoxicillin and tetracycline as an indicator of multidrug resistance (MDR) in animal health. METHODS: Escherichia coli isolates collected between 2012 and 2016 by the French surveillance network for antimicrobial resistance in diseased animals (RESAPATH) were analyzed. The proportions of MDR isolates and the proportions of isolates presenting co-resistance to amoxicillin and tetracycline were calculated for seven animal species (cattle, horse, dog, swine, poultry, duck, and turkey). The degree of agreement between these two proportions was estimated by calculating the kappa value. RESULTS: In total, 55,904 isolates were analyzed. MDR proportions were variable among animal species, ranging from 21.9% [20.2; 23.7] in horses to 56.0% [55.4; 56.7] in cattle. A similar situation was observed for proportions of isolates with co-resistance to amoxicillin and tetracycline, with the highest value for cattle 65.0% [64.3; 65.6]. This co-resistance was also most often associated with resistance to other antibiotics, regardless of the animal species considered. Comparative analysis showed substantial agreement between MDR and this co-resistance, with a kappa value of 0.75, all animal species considered. CONCLUSION: Given the widespread use of penicillins and tetracyclines in animal health, co-resistance to amoxicillin and tetracycline could be an efficient indicator of MDR in E. coli isolates. Based on a specific resistance profile and not an arbitrary number of resistances compared with MDR, this potential indicator is also precise, convenient and suitable for routine use.

Antimicrobial resistance of Pasteurella multocida isolated from diseased food-producing animals and pets.

Surveillance of Pasteurella multocida resistance in food-producing animals is essential to guide the first-line treatment of respiratory diseases and to limit economic losses. Since Pasteurella are the most common bacteria isolated from dog and cat bites, this surveillance is also needed to guide treatment in humans in case of bites. The aim of this study was to characterize the phenotypic resistance of P. multocida strains isolated from respiratory infections in animals, including both food-producing animals and pets. Data collected between 2012 and 2017 by the French national surveillance network for antimicrobial resistance referred to as RESAPATH were analyzed. The proportions of resistance to antimicrobials of relevance in veterinary and human medicines were estimated for each animal species. For cattle, resistance trends over the period were investigated using non-linear analysis applied to time-series. In total, 5356 P. multocida isolates were analyzed. Proportions of resistance of P. multocida were almost all below 20% over the period, and, more precisely, all resistance proportions were below 10% for rabbits, sheep and dogs. The highest resistance proportions to enrofloxacin were identified for cattle (4.5%) and dogs (5.2%). Despite its frequent use in livestock, resistance to florfenicol was less than 1% in P. multocida strains, regardless of the animal species considered. Time series analyses revealed continuous increases in resistance to tetracycline, tilmicosin, flumequine and fluoroquinolones in P. multocida strains isolated from cattle. These trends contrast with the decrease in use of antibiotics in cattle in France and with the decrease in resistance observed in E. coli isolated from diseased cattle.

Antimicrobial Usages and Antimicrobial Resistance in Commensal Escherichia coli From Veal Calves in France: Evolution During the Fattening Process.

Extended-Spectrum-Cephalosporin (ESC)-resistant Enterobacteriaceae have widely spread in all settings worldwide. In animals, Extended-Spectrum Beta-Lactamase (ESBL) producers have been frequently identified in veal calves. The objectives of this study were to investigate the trends in the ESBL load and antimicrobial resistance (AMR) proportions, and antimicrobial usages (AMU) in veal calves during the fattening process. Ten fattening farms were selected and 50 animals per farm were sampled. AMR was assessed in bacteria from the dominant flora (collected on non-selective MacConckey agar) and in ESBL/AmpC-carrying bacteria from the subdominant flora (selected on ChromID ESBL selective plates) upon arrival and 5-6 months later before slaughter. The number and types of treatments during fattening were also collected. Rates of ESBL-producing E. coli from the subdominant flora significantly decreased in all farms (arrival: 67.7%; departure: 20.4%) whereas rates of multidrug-resistant E. coli from the dominant flora have significantly increased (arrival: 60.2%; departure: 67.2%; p = 0.025). CTX-M-1 was the most frequently identified ESBL enzyme (arrival: 59.3%; departure: 52.0%). The plasmid-mediated mcr-1 gene was also identified occasionally. In parallel, levels of resistances to non-critically important antimicrobials were already high upon arrival but have still further increased over time until slaughter. Our study also highlighted that if only ESBL-producing isolates were monitored, it might have led to a partial (and partly false) picture of AMR rates globally decreasing during the fattening period. The mean number of antimicrobial treatments per calf (NTPC) was 8.75 but no association between AMU and AMR was evidenced. Most ESBL producers were clonally unrelated suggesting multiple sources and not cross-contaminations among calves during transportation. Feeding milk containing antimicrobial residues to veal calves is hypothesized to explain the high ESBL loads in animals at the entrance on farms.

Why do veterinarians ask for antimicrobial susceptibility testing? A qualitative study exploring determinants and evaluating the impact of antibiotic reduction policy.

For public health reasons, increasing attention has focused on more rational use of antimicrobials in farm animals. Guidance concerning the prescription of antibiotics and antimicrobial susceptibility testing (antibiograms in this case) are beneficial tools to help control the development of antimicrobial resistance. Nevertheless, even though there are already several qualitative studies analysing the determinants of antimicrobial prescription and use in veterinary medicine, little is known about decision-making concerning the use of antibiograms. The aim of this study was to provide a better understanding of veterinarians' motivations and role-players' influence concerning the choice of whether to ask for an antibiogram in the bovine, porcine, poultry and equine sectors in France. We concurrently evaluated the impact of a new French decree (2016) requiring an antibiogram before certain critically important antimicrobial agents can be used in veterinary medicine. Semi-structured interviews with veterinarians were conducted in France. Thematic analysis was used to analyse transcripts. In all, we surveyed 66 veterinarians. Use of antibiograms in veterinary medicine was multifactorial - 46 factors grouped into 11 categories were identified - and differed between animal sectors: use was almost systematic in poultry, frequent in pigs and rare in both the bovine and equine sectors. The decree has not increased the use of antibiograms but has induced a change in prescriptions due to field constraints and the time needed to obtain the results of antibiograms. Respondents see the decree as an aid in promoting responsible and rational use of antibiotics, fostering the use of alternatives. Our findings provide the basis of veterinarians' position regarding antibiogram use and antimicrobial resistance, pointing out levers to facilitate the use of antibiograms in veterinary medicine (for example communication on the benefits of this test and external financial support). Furthermore, the evaluation of the impact of the decree aimed at reducing the use of critically important antibiotic highlights key factors for a successful change in regulations, such as advance planning, precise and adapted communication, and demonstration of the measure's legitimacy. These results will be useful in guiding representative veterinary bodies and regulatory authorities during their decision-making, communication, and policy and regulation choices to combat antimicrobial resistance.

Comparative epidemiology of E. coli resistance to third-generation cephalosporins in diseased food-producing animals.

Categorized by WHO as critically important antibiotics, third-generation cephalosporins (3GCs) are one of the latest therapeutic alternatives to fight severe infectious diseases in humans. Some antibiotics belonging to this class are prescribed to treat food-producing animals in specific pathological contexts. Preserving the effectiveness of 3GCs requires characterization and careful monitoring of 3GCs resistance and the identification and implementation of measures that can limit this antimicrobial resistance (AMR). Here, we characterized the 3GCs resistance in Escherichia coli isolated from diseased animals. Using data collected from broilers, hens, calves, piglets, sows, turkeys and ducks between 2006 and 2016 by the French surveillance network of AMR in pathogenic bacteria of animal origin (called RESAPATH), we investigated the dynamics of resistance to 3GCs. Our non-linear analysis applied to time series showed that the evolution of E. coli resistance to 3GCs is specific to each animal category. From 2006 to 2010, resistance to 3GCs increased for most animal categories. We observed peaks of high-level of resistance for hens (21.5% in 2010) and broilers (26.7% in 2011), whereas trends stayed below 10% for the other animal categories throughout the study period. Resistance later decreased and, since 2014, 3GCs resistance has dropped below 10% for all animal categories. The parallel between trends and measures to limit AMR over the period shed lights on the impact of practices changes, public policies (EcoAntibio Plan) and sector-led initiatives (moratorium in swine sector). Finally, they highlight the usefulness and importance of AMR surveillance networks in animal health, such as RESAPATH.

An optimal cut-off point for the calving interval may be used as an indicator of bovine abortions.

The bovine abortion surveillance system in France aims to detect as early as possible any resurgence of bovine brucellosis, a disease of which the country has been declared free since 2005. It relies on the mandatory notification and testing of each aborting cow, but under-reporting is high. This research uses a new and simple approach which considers the calving interval (CI) as a "diagnostic test" to determine optimal cut-off point c and estimate diagnostic performance of the CI to identify aborting cows, and herds with multiple abortions (i.e. three or more aborting cows per calving season). The period between two artificial inseminations (AI) was considered as a "gold standard". During the 2006-2010 calving seasons, the mean optimal CI cut-off point for identifying aborting cows was 691 days for dairy cows and 703 days for beef cows. Depending on the calving season, production type and scale at which c was computed (individual or herd), the average sensitivity of the CI varied from 42.6% to 64.4%; its average specificity from 96.7% to 99.7%; its average positive predictive value from 27.6% to 65.4%; and its average negative predictive value from 98.7% to 99.8%. When applied to the French bovine population as a whole, this indicator identified 2-3% of cows suspected to have aborted, and 10-15% of herds suspected of multiple abortions. The optimal cut-off point and CI performance were consistent over calving seasons. By applying an optimal CI cut-off point to the cattle demographics database, it becomes possible to identify herds with multiple abortions, carry out retrospective investigations to find the cause of these abortions and monitor a posteriori compliance of farmers with their obligation to report abortions for brucellosis surveillance needs. Therefore, the CI could be used as an indicator of abortions to help improve the current mandatory notification surveillance system.