Qualitative risk assessment of homogeneity, stability, and residual concentrations of antimicrobials in medicated feed and drinking water in pig rearing.

BACKGROUND: Despite the common use of oral group treatment in pig rearing, the magnitude of the factors influencing the homogeneity and stability of antimicrobial drugs in medicated feed and medicated drinking water are largely unknown, as well as the residual concentrations of the drugs after the end of the treatment. RESULTS: This study presents a qualitative risk assessment to estimate the magnitude of the risks for reduced homogeneity and stability, and increased residual concentrations of antimicrobial drugs in medicated feed and drinking water on the farm. Risk assessment was done using a questionnaire and farm visits (n = 52), combined with a second questionnaire, and concentrations of amoxicillin and doxycycline measured in medicated feed and water samples, each collected on 10 farms. For medicated feed, the duration of storage in the silo did not show to influence the concentration levels in a consistent trend, while the treatment duration had a low to negligible effect. A moderate to high risk was found caused by human error when preparing the medicated feed on the farm. Purchased medicated feed greatly reduces the risk of human error and drugs remain stable during the duration of treatment, while the risk of residual concentrations after the end of the treatment was estimated to be low to moderate. The feed intake variability was identified as a moderate to high risk factor. For medicated drinking water, the type of dosing pump, age of pre-solution, and human errors during the preparation of the pre-solution present a moderate to high risk on homogeneity and stability. Precipitation of the active substance in the absence of a stirrer in a drinking water tank was shown to be a low to moderate risk factor for residues after treatment. Waterline length had a weak correlation with the concentrations of the antimicrobials, while a moderate to high influence was detected for the water intake by the pigs. CONCLUSIONS: A considerable variation in drug concentration in both medicated feed and medicated drinking water was detected depending on their preparation. Therefore, it is important to know which factors influence the homogeneity and stability, and the residual concentrations after treatment.

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.

Biosecurity and antimicrobial use in broiler farms across nine European countries: toward identifying farm-specific options for reducing antimicrobial usage.

Broiler chickens are among the main livestock sectors worldwide. With individual treatments being inapplicable, contrary to many other animal species, the need for antimicrobial use (AMU) is relatively high. AMU in animals is known to drive the emergence and spread of antimicrobial resistance (AMR). High farm biosecurity is a cornerstone for animal health and welfare, as well as food safety, as it protects animals from the introduction and spread of pathogens and therefore the need for AMU. The goal of this study was to identify the main biosecurity practices associated with AMU in broiler farms and to develop a statistical model that produces customised recommendations as to which biosecurity measures could be implemented on a farm to reduce its AMU, including a cost-effectiveness analysis of the recommended measures. AMU and biosecurity data were obtained cross-sectionally in 2014 from 181 broiler farms across nine European countries (Belgium, Bulgaria, Denmark, France, Germany, Italy, the Netherlands, Poland and Spain). Using mixed-effects random forest analysis (Mix-RF), recursive feature elimination was implemented to determine the biosecurity measures that best predicted AMU at the farm level. Subsequently, an algorithm was developed to generate AMU reduction scenarios based on the implementation of these measures. In the final Mix-RF model, 21 factors were present: 10 about internal biosecurity, 8 about external biosecurity and 3 about farm size and productivity, with the latter showing the largest (Gini) importance. Other AMU predictors, in order of importance, were the number of depopulation steps, compliance with a vaccination protocol for non-officially controlled diseases, and requiring visitors to check in before entering the farm. K-means clustering on the proximity matrix of the final Mix-RF model revealed that several measures interacted with each other, indicating that high AMU levels can arise for various reasons depending on the situation. The algorithm utilised the AMU predictive power of biosecurity measures while accounting also for their interactions, representing a first step toward aiding the decision-making process of veterinarians and farmers who are in need of implementing on-farm biosecurity measures to reduce their AMU.

A case of clubbed down syndrome in broilers.

This study presents a case of clubbed down syndrome in conventional broilers. During the first week of life, severe growth retardation was observed in approximately 25% of the flock. The growth-retarded chicks weighed only 45 g and showed a typical feather disorder which was most apparent on their abdomen and was defined in literature as typical for clubbed down syndrome. Necropsies, histology, biochemical analysis of blood and liver samples, serology and different PCR tests were performed in broilers to assess the aetiology of the clinical signs that were present in the affected broiler farm. Because of the suspicion of a possible link with the broiler-breeder farms, different investigations including serology, PCR and feed analysis were also performed on these farms. The results suggest that an accidentally excessive amount of calcium and iron in the feed of broiler-breeders, 3 weeks prior to first clinical signs in broilers, led to the development of clubbed down in the offspring, because of a relative Zn-deficiency in broiler-breeders and an absolute Zn-deficiency in the hatching eggs that were produced during this period. This appeared to be a reversible process as no clinical signs were observed in younger offspring of these broiler-breeders after they had consumed more of the new batch of feed. A potential involvement of Astrovirus could not be completely ruled out. This study demonstrates the importance of correct mineral concentrations in broiler-breeder feed and the impact it can have on the development of the offspring.

A study on risk factors for macroscopic gut abnormalities in intensively reared broiler chickens.

Intensification of broiler production has coincided with an increase in enteric disorders. Enteric syndromes of unknown aetiology are often associated with an increased feed conversion ratio and are given the general term "dysbiosis". Despite the importance of dysbiosis, information on factors contributing to this condition are scarce. Therefore, the aim of this study was to describe dysbiosis in broilers (Ross 308) during one production round and to identify risk factors. Fifteen farms in Flanders (Belgium) were followed up, with visits at days 10, 17, 20, 24 and 28 of production. At every visit, 10 random birds were inspected for footpad lesions, hock burns and breast blisters. Also, coccidiosis and enteric abnormalities were scored after necropsy. A gut appearance score (GAS) was given based on 10 macroscopically visible parameters, where a higher GAS equalled more enteric abnormalities. Footpad lesions were seen in 14 farms and increased in prevalence with the age of the birds. Hock burns were seen less frequently, and no breast blisters were detected. Eimeria acervulina lesions were most frequently observed, followed by E. maxima and E. tenella lesions. The average GAS increased from day 10 until day 20. The strong correlations between the GAS at days 10, 17 and 20 indicate that prevalence of gut abnormalities at day 10 can be predictive for scores later on. A higher amount of intestinal defects was seen in older female birds, in the presence of a conceivable E. tenella infection and at farms with a higher productivity and sanitary status.RESEARCH HIGHLIGHTSGut lesions found in young broilers can predict further development of defects.Caecal lesions showed the strongest correlation with GAS in a multivariate model.

Risk factors for poor health and performance in European broiler production systems.

BACKGROUND: Conventional broilers are currently one of the most efficient protein converters. Although decades of progress in genetic selection and feed formulation have lead to high standards of efficient broiler production, still a lot of variability is found between farms and between successive flocks. The aim of this study was to investigate risk- and/or protective factors for poor health and performance in conventional broiler-farms in Europe by developing eight multivariable linear mixed models. Three different models were used to investigate mortality (overall, first week, after first week), three models for performance variables (growth, feed conversion, European production index) and two models were related to slaughterhouse data (i.e. dead on arrival and condemnation rate). RESULTS: Several factors related to management and housing were significantly associated with health and performance of broilers. The following factors were associated with increased mortality: floor quality, neonatal septicemia, ventilation type and other professional activities of the farmer. The factors associated with performance were chick sex, coccidiosis infections, necrotic enteritis, dysbacteriosis, light intensity adaptations, ventilation type, comparing daily flock results with previous flock results by farmer, daily check of feed and water system and type of feed. For dead on arrival three risk factors were identified i.e. daily growth, type of light adaptation and type of drinkers system. For condemnation rate seven risk factors were found, i.e. type of drinking system, daily growth, feed withdrawal time, type of ventilation, house size, septicemia after seven days and type of feed. CONCLUSIONS: These results imply that a multifactorial approach is required with adaptations involving both improvements in management, housing, health programs and an increasing level of professionalism of the farmer in order to improve broiler performance and health.

Quantitative and qualitative analysis of antimicrobial usage at farm and flock level on 181 broiler farms in nine European countries.

OBJECTIVES: To control the emerging threat of antimicrobial resistance, international policy appeals for appropriate monitoring of antimicrobial usage (AMU) at supranational, species and farm level. The aim of this study was to quantify AMU in broilers at farm and flock level in nine European countries. METHODS: Antimicrobial treatment data of one flock and purchased antimicrobials over one year were collected at 181 European broiler farms. Afterwards AMU was quantified using treatment incidence (TI) per 100 days based on Defined Daily Dose (DDDvet), Defined Course Dose (DCDvet) or Used Daily Dose (UDDvet) values. Total AMU at flock level was obtained by summing the TIDDDvet of all treatments in the sampled flock (TIDDDvetFl*). RESULTS: The median TIDDDvetFl* was 9.0 (95% CI 5.5-10.8), meaning that broilers were treated with antimicrobials during 9% of their rearing period. TIDDDvetFl* varied considerably within and between countries. However, in every country at least one untreated flock was present. Average TIDDDvetFl* at country level ranged from 3.3 to 36.7. Polymyxins, extended-spectrum aminopenicillins and fluoroquinolones were the most used antimicrobials, accounting for 26%, 26% and 18% of total AMU, respectively. Twenty-six percent of the farms started a treatment on day 1 of production, and 49% of overall AMU was administered within the first week. CONCLUSIONS: Results show that rearing broilers without AMU is feasible. However, a huge variation in AMU in terms of amount, moment of administration and antimicrobial classes was observed. This shows that there is still ground to be covered when it comes to AMU on broiler farms.

Quantitative and qualitative analysis of antimicrobial usage patterns in 180 selected farrow-to-finish pig farms from nine European countries based on single batch and purchase data.

OBJECTIVES: Farm-level quantification of antimicrobial usage (AMU) in pig farms. METHODS: In a cross-sectional study, AMU data on group treatments administered to a single batch of fattening pigs from birth to slaughter (group treatment data) and antimicrobials purchased during 1 year (purchase data) were collected at 180 pig farms in nine European countries. AMU was quantified using treatment incidence (TI) based on defined (DDDvet) and used (UDDvet) daily doses and defined (DCDvet) and used (UCDvet) course doses. RESULTS: The majority of antimicrobial group treatments were administered to weaners (69.5% of total TIDDDvet) followed by sucklers (22.5% of total TIDDDvet). AMU varied considerably between farms with a median TIDDDvet of 9.2 and 7.1 for a standardized rearing period of 200 days based on group treatment and purchase data, respectively. In general, UDDvet and UCDvet were higher than DDDvet and DCDvet, respectively, suggesting that either the defined doses were set too low or that group treatments were often dosed too high and/or administered for too long. Extended-spectrum penicillins (31.2%) and polymyxins (24.7%) were the active substances most often used in group treatments, with the majority administered through feed or water (82%). Higher AMU at a young age was associated with higher use in older pigs. CONCLUSIONS: Collecting farm-level AMU data of good quality is challenging and results differ based on how data are collected (group treatment data versus purchase data) and reported (defined versus used daily and course doses).

The antimicrobial resistome in relation to antimicrobial use and biosecurity in pig farming, a metagenome-wide association study in nine European countries.

OBJECTIVES: Previous studies in food-producing animals have shown associations between antimicrobial use (AMU) and resistance (AMR) in specifically isolated bacterial species. Multi-country data are scarce and only describe between-country differences. Here we investigate associations between the pig faecal mobile resistome and characteristics at the farm-level across Europe. METHODS: A cross-sectional study was conducted among 176 conventional pig farms from nine European countries. Twenty-five faecal samples from fattening pigs were pooled per farm and acquired resistomes were determined using shotgun metagenomics and the Resfinder reference database, i.e. the full collection of horizontally acquired AMR genes (ARGs). Normalized fragments resistance genes per kilobase reference per million bacterial fragments (FPKM) were calculated. Specific farm-level data (AMU, biosecurity) were collected. Random-effects meta-analyses were performed by country, relating farm-level data to relative ARG abundances (FPKM). RESULTS: Total AMU during fattening was positively associated with total ARG (total FPKM). Positive associations were particularly observed between widely used macrolides and tetracyclines, and ARGs corresponding to the respective antimicrobial classes. Significant AMU-ARG associations were not found for ß-lactams and only few colistin ARGs were found, despite high use of these antimicrobial classes in younger pigs. Increased internal biosecurity was directly related to higher abundances of ARGs mainly encoding macrolide resistance. These effects of biosecurity were independent of AMU in mutually adjusted models. CONCLUSIONS: Using resistome data in association studies is unprecedented and adds accuracy and new insights to previously observed AMU-AMR associations. Major components of the pig resistome are positively and independently associated with on-farm AMU and biosecurity conditions.

Associations between antimicrobial use and the faecal resistome on broiler farms from nine European countries.

OBJECTIVES: To determine associations between farm- and flock-level antimicrobial usage (AMU), farm biosecurity status and the abundance of faecal antimicrobial resistance genes (ARGs) on broiler farms. METHODS: In the cross-sectional pan-European EFFORT study, conventional broiler farms were visited and faeces, AMU information and biosecurity records were collected. The resistomes of pooled faecal samples were determined by metagenomic analysis for 176 farms. A meta-analysis approach was used to relate total and class-specific ARGs (expressed as fragments per kb reference per million bacterial fragments, FPKM) to AMU (treatment incidence per DDD, TIDDDvet) per country and subsequently across all countries. In a similar way, the association between biosecurity status (Biocheck.UGent) and the resistome was explored. RESULTS: Sixty-six (38%) flocks did not report group treatments but showed a similar resistome composition and roughly similar ARG levels to antimicrobial-treated flocks. Nevertheless, we found significant positive associations between ß-lactam, tetracycline, macrolide and lincosamide, trimethoprim and aminoglycoside antimicrobial flock treatments and ARG clusters conferring resistance to the same class. Similar associations were found with purchased products. In gene-level analysis for ß-lactams and macrolides, lincosamides and streptogramins, a significant positive association was found with the most abundant gene clusters blaTEM and erm(B). Little evidence was found for associations with biosecurity. CONCLUSIONS: The faecal microbiome in European broilers contains a high diversity of ARGs, even in the absence of current antimicrobial selection pressure. Despite this, the relative abundance of genes and the composition of the resistome is positively related to AMU in European broiler farms for several antimicrobial classes.

Limited association between disinfectant use and either antibiotic or disinfectant susceptibility of Escherichia coli in both poultry and pig husbandry.

BACKGROUND: Farm disinfectants are widely used in primary production, but questions have been raised if their use can select for antimicrobial resistance. The present study examined the use of disinfectants in poultry and pig husbandry and its contribution to the antibiotic and disinfectant susceptibility of Escherichia coli (E. coli) strains obtained after cleaning and disinfection. On those field isolates antibiotic susceptibility was monitored and susceptibility to commonly used active components of farm disinfectants (i.e. glutaraldehyde, benzalkoniumchloride, formaldehyde, and a formulation of peracetic acid and hydrogen peroxide) was tested. RESULTS: This study showed a high resistance prevalence (> 50%) for ampicillin, sulfamethoxazole, trimethoprim and tetracycline for both production animal categories, while for ciprofloxacin only a high resistance prevalence was found in broiler houses. Disinfectant susceptibility results were homogenously distributed within a very small concentration range. Furthermore, all E. coli strains were susceptible to in-use concentrations of formaldehyde, benzalkoniumchloride and a formulation of peracetic acid and hydrogen peroxide, indicating that the practical use of disinfectants did not select for disinfectant resistance. Moreover, the results showed no indications for the selection of antibiotic resistant bacteria through the use of disinfectants in agricultural environments. CONCLUSION: Our study suggests that the proper use of disinfectants in agricultural environments does not promote antibiotic resistance nor reduce E. coli disinfectant susceptibility.

The effect of a commercial competitive exclusion product on the selection of enrofloxacin resistance in commensal E. coli in broilers.

The effect of a competitive exclusion product (Aviguard®) on the selection of fluoroquinolone resistance in poultry was assessed in vivo in the absence or presence of fluoroquinolone treatment. Two experiments using a controlled seeder-sentinel animal model (2 seeders: 4 sentinels per group) with one-day-old chicks were used. For both experiments, as soon as the chicks were hatched, the birds of two groups were administered Aviguard® and two groups were left untreated. Three days later, all groups were inoculated with an enrofloxacin-susceptible commensal E. coli strain. Five days after hatching, two birds per group were inoculated with either a bacteriologically fit or a bacteriologically non-fit enrofloxacin-resistant commensal E. coli strain. In experiment 2, all groups were orally treated for three consecutive days (days 8-10) with enrofloxacin. Throughout the experiments, faecal excretion of all inoculated E. coli strains was determined on days 2, 5, 8, 11, 18 and 23 by selective plating (via spiral plater). Linear mixed models were used to assess the effect of Aviguard® on the selection of fluoroquinolone resistance. The use of Aviguard® (P < 0.01) reduced the excretion of enrofloxacin-resistant E. coli when no enrofloxacin treatment was administered. However, this beneficial effect disappeared (P = 0.37) when the birds were treated with enrofloxacin. Similarly, bacterial fitness of the enrofloxacin-resistant E. coli strain used for inoculation had an effect (P < 0.01) on the selection of enrofloxacin resistance when no treatment was administered, whereas this effect was no longer present when enrofloxacin was administered (P = 0.70). Thus, enrofloxacin treatment cancelled the beneficial effects from administrating Aviguard® in one-day-old broiler chicks and resulted in an enrofloxacin-resistant flora. RESEARCH HIGHLIGHTS The effect of Aviguard® on the selection of enrofloxacin resistance was assessed in vivo. Without enrofloxacin, Aviguard® reduced the selection of enrofloxacin resistance. When enrofloxacin was administered, it cancelled the beneficial effect of Aviguard®.

Inflammatory bowel disease (IBD) in horses: a retrospective study exploring the value of different diagnostic approaches.

BACKGROUND: Diagnosing IBD in horses is challenging and requires a multimodal approach, since no conclusive diagnostic test is available. The objectives of this study were to provide an overview of population characteristics, results of applied diagnostic tests, treatment modalities and outcome in a large group of horses thought to have IBD and that were presented to four large equine referral hospitals, and to provide an exploratory investigation of possible associations between results of applied diagnostic tests, applied treatment modalities and outcome. A retrospective case series was performed across four large equine referral hospitals. Seventy-eight horses, thought to have IBD were included. Case history, clinical findings, diagnostic test results including oral glucose tolerance test (OGTT) and enteral biopsies (both duodenal and rectal), applied therapy and outcome were studied. A Chi-Square test was used to identify associations between results of diagnostic tests, treatment and outcome. P-values < 0.05 were considered significant. RESULTS: Lethargy, diarrhoea, recurrent colic and weight loss were recorded in respectively 21,8%; 14,1%; 28,2% and 78,2% of cases. Over 70% of horses thought to have IBD had an abnormal OGTT. Only weight loss was significantly associated with aberrant enteral biopsy results, but not with abnormal OGTT results or low blood total protein. There was no association between an aberrant biopsy result and a disturbed OGTT. There was no association between either OGTT results or enteral biopsy results and a low blood total protein content, presence of gastric ulcer disease or an aberrant endoscopic aspect of the duodenal entrance. CONCLUSIONS: Weight loss is a highly prevalent symptom in IBD suspected horses. Enteral biopsies may be a useful diagnostic aid in the work-up of horses thought to suffer from IBD, however further research is required to demonstrate their true diagnostic value. Until more standardized scientific research is available, one should be careful with the interpretation of enteral biopsy results There is a need for better standardization of enteral biopsy procedures and the histopathological scoring of biopsies.

Studying the effect of administration route and treatment dose on the selection of enrofloxacin resistance in commensal Escherichia coli in broilers.

Objectives: Factors potentially contributing to fluoroquinolone resistance selection in commensal Escherichia coli strains in poultry were studied through a series of in vivo experiments. The effect of the initial prevalence of enrofloxacin resistance in the E. coli gut microbiota, effect of the bacterial fitness of the enrofloxacin-resistant strain and effect of treatment with enrofloxacin (effect of dose and effect of route of administration) were assessed. Methods: Four in vivo studies with broiler chickens were performed. Right after hatching, the chicks were inoculated with either a bacteriologically fit or a bacteriologically non-fit fluoroquinolone-resistant strain as either a minority or the majority of the total E. coli population. Six days later, the chicks were treated for three consecutive days either orally or parenterally and using three different doses (under-, correct- and over-dose) of enrofloxacin. The faecal shedding of E. coli strains was quantified by plating on agar plates either supplemented or not supplemented with enrofloxacin. Linear mixed models were used to assess the effect of the aforementioned variables on the selection of enrofloxacin resistance. Results: The factors that significantly contributed were treatment ( P < 0.001), bacterial fitness of the resistant donor strain ( P < 0.001), administration route ( P = 0.052) and interactions between bacterial fitness and administration route ( P < 0.001). Conclusions: In the currently used models, fluoroquinolone resistance selection was influenced by treatment, bacterial fitness of the inoculation strain and administration route. The use of oral treatment seems to select more for fluoroquinolone resistance, particularly in the model where a non-fit strain was used for inoculation.

Residues of chlortetracycline, doxycycline and sulfadiazine-trimethoprim in intestinal content and feces of pigs due to cross-contamination of feed.

BACKGROUND: Cross-contamination of feed with low concentrations of antimicrobials can occur at production, transport and/or farm level. Concerns are rising about possible effects of this contaminated feed on resistance selection in the intestinal microbiota. Therefore, an experiment with pigs was set up, in which intestinal and fecal concentrations of chlortetracycline (CTC), doxycycline (DOX) and sulfadiazine-trimethoprim (SDZ-TRIM) were determined after administration of feed containing a 3 % carry-over level of these antimicrobials. RESULTS: The poor oral bioavailability of tetracyclines resulted in rather high concentrations in cecal and colonic content and feces at steady-state conditions. A mean concentration of 10 mg/kg CTC and 4 mg/kg DOX in the feces was reached, which is higher than concentrations that were shown to cause resistance selection. On the other hand, lower mean levels of SDZ (0.7 mg/kg) and TRIM (< limit of detection of 0.016 mg/kg) were found in the feces, corresponding with the high oral bioavailability of SDZ and TRIM in pigs. CONCLUSIONS: The relation between the oral bioavailability and intestinal concentrations of the tested antimicrobials, may be of help in assessing the risks of cross-contaminated feed. However, future research is needed to confirm our results and to evaluate the effects of these detected concentrations on resistance selection in the intestinal microbiota of pigs.

Effect of sow vaccination against porcine circovirus type 2 (PCV2) on virological profiles in herds with or without PCV2 systemic disease.

We investigated porcine circovirus type 2 (PCV2) virological profiles in herds affected (PCVAD-AH, n = 5) or non-affected (PCVAD-NAH, n = 4) by PCV2-associated diseases (PCVAD), before and after 1 y of PCV2 gilt and sow vaccination. Fresh feces from the floor (5 pens/age/farm) and 5 blood samples (1/pen) were collected at 3, 9, 15, 21 wk. Individual feces and blood samples were collected from 5 gilts and 15 sows. Sampling was repeated 1 y after vaccination. Quantitative PCR on feces, PCV2 antibodies in blood serum and cell-mediated immunity were investigated. Before vaccination, pigs of PCVAD-AH had higher viral load in feces (9 and 15 wk), lower IgG and higher IgM (3 wk) and lower lymphocyte counts (9 and 15 wk) suggesting immunosuppression. Vaccination reduced viral load in growers, increased IgG (3 wk) suggesting improved maternal immunity, reduced IgM (3 wk), increased total antibody titers in sows and increased CD79a cells in the pigs.

Effet de la vaccination des truies contre le circovirus porcin de type 2 (PCV2) sur les profils virologiques des troupeaux atteints ou non de la maladie systémique PCV2. Nous avons fait une enquête sur les profils virologiques du circovirus porcin de type 2 (PCV2) dans les troupeaux affectés (PCVAD-AH, n = 5) ou non affectés (PCVAD-NAH, n = 4) par les maladies associées au PCV2 (MAPCV), 1 an avant et 1 an après la vaccination des cochettes et des truies contre le PCV2. Des fèces fraîches sur le plancher (5 enclos/âge/ferme) et 5 échantillons de sang (1/enclos) ont été prélevés à 3, 9, 15 et 21 semaines. Des fèces individuelles et des échantillons sanguins ont été préIevés auprès de 5 cochettes et de 15 truies. L'échantillonnage a été répété 1 an après la vaccination. La RCP quantitative sur les fèces, les anticorps de PCV2 dans le sérum sanguin et l'immunité à médiation cellulaire ont fait l'objet d'une enquête. Avant la vaccination, les porcs de PCVAD-AH présentaient une charge virale supérieure dans les fèces (à 9 et à 15 semaines), une IgG inférieure et une IgM supérieure (à 3 semaines) ainsi qu'une numération inférieure des lymphocytes (à 9 et à 15 semaines) suggérant l'immunosuppression. La vaccination a réduit la charge virale chez les porcs en croissance, a augmenté les IgG (à 3 semaines) suggérant une immunité maternelle améliorée, a réduit les IgM (à 3 semaines), a augmenté le total des titres d'anticorps chez les truies et a augmenté les cellules CD79a chez les porcs.(Traduit par Isabelle Vallières).

Assigning defined daily doses animal: a European multi-country experience for antimicrobial products authorized for usage in pigs.

OBJECTIVES: To establish a consensus defined daily dose animal (DDDA) for each active substance (AS) and administration route for porcine veterinary antimicrobial products authorized in four European countries, thus allowing cross-country quantification and comparison of antimicrobial usage data. METHODS: All veterinary antimicrobial products authorized for porcine use in Belgium, France, Germany and Sweden were listed for each administration route. First, separate DDDAs for each product were defined based on the recommended dosing for the main indication. Second, a consensus DDDA was established by taking the mean of the DDDAs for each product within a certain category of AS plus administration route. RESULTS: One-hundred-and-fifty-nine, 240, 281 and 50 antimicrobial products were licensed in Belgium, France, Germany and Sweden, respectively, in February 2013. Large variations were observed for dosage and treatment duration recommendations between products and between countries for the same ASs. Only 6.8% of feed/water and 29.4% of parenteral AS groups had the same recommended dosage in the four countries. CONCLUSIONS: This study presents a consensus DDDA list for use in the quantification and comparison of antimicrobial consumption. Four major recommendations have been formulated: (i) urgent need for harmonization of authorization and recommended summary of product characteristics (SPC) dosages; (ii) expand the developed preliminary DDDA list to include all authorized veterinary medicinal products in all EU member states and for all (food-producing) animal species; (iii) improved accessibility of country-specific SPC data would be preferable; and (iv) statement of the 'long-acting' duration of a product in the SPC.

Interactions of highly and low virulent Flavobacterium columnare isolates with gill tissue in carp and rainbow trout.

The interactions of Flavobacterium columnare isolates of different virulence with the gills of carp (Cyprinus carpio L.) and rainbow trout (Oncorhynchus mykiss Walbaum) were investigated. Both fish species were exposed to different high (HV) or low virulence (LV) isolates and sacrificed at seven predetermined times post-challenge. Histopathological and ultrastructural examination of carp and rainbow trout inoculated with the HV-isolate disclosed bacterial invasion and concomitant destruction of the gill tissue, gradually spreading from the filament tips towards the base, with outer membrane vesicles surrounding most bacterial cells. In carp, 5-10% of the fish inoculated with the LV-isolate became moribund and their gill tissue displayed the same features as described for the HV-isolate, albeit to a lesser degree. The bacterial numbers retrieved from the gill tissue were significantly higher for HV- compared to LV-isolate challenged carp and rainbow trout. TUNEL-stained and caspase-3-immunostained gill sections demonstrated significantly higher apoptotic cell counts in carp and rainbow trout challenged with the HV-isolate compared to control animals. Periodic acid-Schiff/alcian blue staining demonstrated a significantly higher total gill goblet cell count for HV- and LV-isolate challenged compared to control carp. Moreover, bacterial clusters were embedded in a neutral matrix while being encased by acid mucins, resembling biofilm formation. Eosinophilic granular cell counts were significantly higher in the HV-isolate compared to LV-isolate inoculated and control carp. The present data indicate a high colonization capacity, and the destructive and apoptotic-promoting features of the HV-isolate, and point towards important dynamic host mucin-F. columnare interactions warranting further research.

Correlation between veterinary antimicrobial use and antimicrobial resistance in food-producing animals: a report on seven countries.

OBJECTIVES: To evaluate correlations between antimicrobial use and the prevalence of resistance in commensal Escherichia coli isolates from pigs, poultry and cattle, using data from publicly available national or international reports from seven European countries. METHODS: The link between the quantities of different classes of antimicrobials administered to food-producing animals per country (expressed in mg/population correction unit) and the prevalence of resistance to the different antimicrobial classes (interpreted by EUCAST epidemiological cut-off values) in E. coli isolates (4831 isolates in total) was assessed by means of polynomial regression analysis and determination of Spearman's rank correlation coefficient. RESULTS: A quadratic regression best fitted the antimicrobial use and antimicrobial resistance data. The coefficient of determination was, in decreasing order, 0.99 for fluoroquinolones and amphenicols, 0.94 for third-generation cephalosporins and sulphonamides, 0.93 for aminopenicillins, 0.86 for fluoroquinolones, 0.81 for streptomycin and 0.80 for gentamicin and tetracycline. Spearman's rank correlation coefficient was 1 for amphenicols, 0.96 for sulphonamides, 0.93 for streptomycin and tetracycline, 0.89 for aminopenicillins, 0.71 for gentamicin and 0.70 for third-generation cephalosporins. CONCLUSIONS: These remarkably high coefficients indicate that, at a national level, the level of use of specific antimicrobials strongly correlates to the level of resistance towards these agents in commensal E. coli isolates in pigs, poultry and cattle. However, data restraints reveal the need for further detail in collection and harmonization of antimicrobial resistance and use data in Europe.

Individual commitment to a group effect: strengths and weaknesses of bovine embryo group culture.

Recently, new culture devices such as Corral and Primo Vision dishes have been designed for the culture of human embryos to allow the combination of group culture plus follow-up of individual embryos. Bovine inseminated oocytes were allocated to Primo Vision dishes, Corral dishes, individual culture or classical group culture. Blastocyst development in Primo Vision dishes was similar to classical group culture (34.3 and 39.0% respectively), and better than Corral dishes or individual culture (28.9 and 28.5% respectively). In Primo Vision dishes, a higher number of 'slow' embryos developed to the blastocyst stage compared with their individually cultured counterparts, while no differences were observed for 'fast' embryos. 'Slow' embryos in a 'standard drop' had a higher chance of becoming a blastocyst compared with individual culture (OR: 2.3), whereas blastulation of 'fast' embryos was less efficient in a 'delayed drop' than in individual culture (OR: 0.3). The number of non-cleaved embryos in Primo Vision dishes did not negatively influence blastocyst development. Likewise, removing non-cleaved embryos (NC removed) and regrouping the cleaved embryos afterwards (ReGR) did not affect blastocyst development and quality compared with group culture in Primo Vision dishes (CTRL, 31.6%, NC removed, 29.3% and ReGR, 29.6%). The experiments revealed that group culture of bovine embryos in Primo Vision dishes is superior to individual culture, primarily because of the higher blastocyst rate achieved by slow embryos. Non-cleaved or arrested embryos do not hamper the ability of co-cultured bovine embryos to reach the blastocyst stage in group culture.