A non-invasive tool to collect small intestine content in post weaning pigs: validation study.

The Capsule for Sampling (CapSa) is an ingestible capsule that collects small intestine content while transiting through the natural digestive pathway. In this study, 14 Swiss Large White pigs weighing less than 12 kg (Category < 12 kg) and 12 weighing between 12 and 20 kg (Category [12-20 kg]) were given two CapSas and monitored for three days. The animals were euthanized for post-mortem sampling, allowing us to directly obtain gut microbiota samples from the gastrointestinal tract. This post-mortem approach enabled a direct comparison between the microbial content from the gut and the samples collected via the CapSas, and it also facilitated precise identification of the CapSas' sampling sites within the gastrointestinal tract. For the category under 12 kg, only 2.3% of the administered CapSas were recovered from the feces. In contrast, in the 12-20 kg category, 62.5% of the CapSas were successfully retrieved from the feces within 48 h. Of these recovered CapSas, 73.3%-equating to 11 capsules from eight pigs-had a pH > 5.5 and were therefore selected for microbiome analysis. Bacterial composition of the CapSas was compared with that of the three segments of the small intestine, the large intestine and feces of the corresponding pig. The results were tested using a PERMANOVA model (Adonis) including sample type as a factor, and then pairwise comparisons were made. The bacterial composition found in the CapSas differed from that of the large intestine and feces (P < 0.01), while it did not differ from the first segment of the small intestine (P > 0.10). This study provides evidence that the CapSa effectively samples the intestinal microbiota from the upper section of the small intestine in post-weaning pigs. Furthermore, it was found that the collection of CapSas could only be successfully achieved in pigs classified within the heavier weight category.

Whole-genome sequencing-based antimicrobial resistance and shedding dynamics of Escherichia coli isolated from calves before and after antimicrobial group treatments.

The fattening of calves is often associated with high antimicrobial use and the selection of antimicrobial resistance (AMR). The objective of this observational longitudinal study was to describe the AMR and strain dynamics, using whole-genome sequencing (WGS), of fecal Escherichia coli in a cohort of 22 calves. All calves received antimicrobial group treatments on Day (D) 1 (oxytetracycline, intramuscularly) and on D4 through D12 (doxycycline, in-feed). Additionally, eight calves received individual parenteral treatments between D7 and D59, including florfenicol, amoxicillin, marbofloxacin, and gamithromycin. Rectal swabs were collected from all calves on D1 (prior to treatment), D2, D9, and D82. The swabs were spread onto Enterobacterales-selective agar, and three E. coli colonies per plate were subjected to WGS. Out of 264 isolates across all calves and sampling times, 80 unique strains were identified, a majority of which harbored genes conferring resistance to tetracyclines, streptomycin, and sulfonamides. The diversity of strains decreased during the in-feed antimicrobial group treatment of the calves. On D82, 90% of isolates were strains that were not isolated at previous sampling times, and the median number per strain of AMR determinants to tetracyclines, florfenicol, ß-lactams, quinolones, or macrolides decreased compared to D9. Additionally, clonal dissemination of some strains represented the main transmission route of AMR determinants. In this study, WGS revealed important variations in strain diversity and genotypic AMR of fecal E. coli over time in calves subjected to group antimicrobial treatments. IMPORTANCE: The continued emergence and spread of antimicrobial resistance (AMR) determinants are serious global concerns. The dynamics of AMR spread and persistence in bacterial and animal host populations are complex and not solely driven by antimicrobial selection pressure. In calf fattening, both antimicrobial use and carriage prevalence of antimicrobial-resistant bacteria are generally recognized as high. This study provides new insights into the short-term, within-farm dynamics and transmission of AMR determinants in Escherichia coli from the dominant fecal flora of calves subjected to antimicrobial group treatments during the rearing period. The diversity of E. coli strains decreased over time, although, in contrast to previous observations in extended-spectrum ß-lactamase-producing Enterobacterales, the predominance of a few clones was not observed. The spread of AMR determinants occurred through the dissemination of clonal strains among calves. The median number per strain of AMR determinants conferring resistance to selected antimicrobials decreased toward the end of the rearing period.

Machine learning algorithms can predict tail biting outbreaks in pigs using feeding behaviour records.

Tail biting is a damaging behaviour that impacts the welfare and health of pigs. Early detection of precursor signs of tail biting provides the opportunity to take preventive measures, thus avoiding the occurrence of the tail biting event. This study aimed to build a machine-learning algorithm for real-time detection of upcoming tail biting outbreaks, using feeding behaviour data recorded by an electronic feeder. Prediction capacities of seven machine learning algorithms (Generalized Linear Model with Stepwise Feature Selection, random forest, Support Vector Machines with Radial Basis Function Kernel, Bayesian Generalized Linear Model, Neural network, K-nearest neighbour, and Partial Least Squares Discriminant Analysis) were evaluated from daily feeding data collected from 65 pens originating from two herds of grower-finisher pigs (25-100kg), in which 27 tail biting events occurred. Data were divided into training and testing data in two different ways, either by randomly splitting data into 75% (training set) and 25% (testing set), or by randomly selecting pens to constitute the testing set. In the first data splitting, the model is regularly updated with previous data from the pen, whereas in the second data splitting, the model tries to predict for a pen that it has never seen before. The K-nearest neighbour algorithm was able to predict 78% of the upcoming events with an accuracy of 96%, when predicting events in pens for which it had previous data. Our results indicate that machine learning models can be considered for implementation into automatic feeder systems for real-time prediction of tail biting events.

Hydrolysable chestnut tannins for reduction of postweaning diarrhea: Efficacy on an experimental ETEC F4 model.

An experimental model for postweaning diarrhea with enterotoxigenic Escherichia coli F4 (ETEC F4) was set up in piglets, and the efficacy of 1% chestnut-tannin extract in preventing diarrhea was subsequently assessed. In a first trial (infection model), 32 Swiss Large White piglets (age: 24 days; average BW: 7.8 ± 0.8 kg) were randomly assigned to two experimental groups (infected [INF], noninfected [NINF]). In a subsequent trial, 72 Swiss Large White piglets (age: 26 days; average BW: 7.4 ± 1.5 kg) were blocked by BW and assigned within block to four experimental groups: NINF-CO: not infected and fed a standard control starter diet (CO); INF-CO: infected and fed the CO diet; NINF-TA: not infected and fed the CO diet supplemented with 1% chestnut extract containing 54% of hydrolysable tannins (TA); and INF-TA: infected and fed the TA diet. Both diets (TA and CO) were formulated to be isocaloric and isoproteic and to meet or surpass the nutritional requirements. In both trials, four days after weaning, piglets assigned to the INF group received an oral suspension of ETEC F4. Fecal score, ETEC shedding in feces (only in trial 2), and growth performance traits were measured for the following 14 days post infection. In both trials, more than 50% of the INF piglets developed diarrhea within six days post infection. Tannins reduced (P < 0.05) the average fecal score, the percentage of piglets in diarrhea, and the duration of diarrhea, whereas feed intake and the average daily gain were unaffected.

Prevention of transmission of Ehrlichia canis by Rhipicephalus sanguineus ticks to dogs treated with a combination of fipronil, amitraz and (S)-methoprene (CERTIFECT®).

The ability of CERTIFECT(®) (a combination of fipronil, amitraz and (S)-methoprene) to prevent transmission of Ehrlichia canis was studied in two groups of eight dogs. One group was treated with CERTIFECT while the other group remained untreated. All dogs were exposed to E. canis-infected Rhipicephalus sanguineus ticks on Days 7, 14, 21 and again on day 28 post-treatment by releasing ticks into the kennels of the dogs to simulate the natural way of infestation. Animals were examined in situ for ticks on Days 9, 16 and 23 and any ticks present were counted and removed on Day 30. The efficacy of CERTIFECT against R. sanguineus was 100%, since no ticks were found on the treated dogs at any time. Clinical examinations (including monitoring body temperature and blood collections for PCR analysis and serology) were performed at intervals throughout the study until Day 56. Five out of 8 untreated control dogs (62.5%) became infected with E. canis, as demonstrated by detection of specific E. canis antibodies and the presence of E. canis DNA in blood samples by PCR. These dogs displayed fever and thrombocytopenia and were rescue-treated with doxycline. None of the 8 dogs treated with CERTIFECT became infected with E. canis, in comparison to the 5/8 control dogs, as confirmed by the lack of specific antibodies and absence of any ehrlichial DNA in blood samples by PCR. CERTIFECT prevented transmission of E. canis and effectively provided protection against monocytic ehrlichiose for at least 4 weeks post treatment.

Efficacy of a novel combination of fipronil, amitraz and (S)-methoprene for treatment and control of tick species infesting dogs in Europe.

Four studies were conducted to show the effectiveness of a novel combination of fipronil, amitraz and (S)-methoprene in a spot-on formulation (CERTIFECT™, Merial Limited, GA, USA) for the therapeutic and preventive control of Ixodid tick species affecting dogs in Europe: Ixodes ricinus, Dermacentor reticulatus and Rhipicephalus sanguineus. In each, untreated control dogs were compared to others treated with the novel combination. All dogs were infested with 50 adult, unfed ticks prior to treatment and at 7-day intervals after treatment. Ticks on all dogs were counted at 18, 24 and 48 h after treatment (therapeutic efficacy) or infestation (preventive efficacy). Therapeutic efficacy of fipronil, amitraz and (S)-methoprene was excellent as shown by significant (p<0.05) and greater than 97% and up to 100% reductions in the 48 h tick counts and significant (p<0.05) detachment/death of ticks evident at 18-24h after treatment for all three tick species. Preventive efficacy was demonstrated by significant (p<0.05) and greater than 93% and up to 100% reductions in tick counts at 48 h after repeat infestations out to 35 days after treatment for I. ricinus and out to 42 days after treatment for D. reticulatus and R. sanguineus. The time to substantial disruption of establishment of new tick infestations after treatment was less than 18-24h and was maintained for up to 28 days after treatment of I. ricinus and D. reticulatus infestations, and 4h to at most 18 h and maintained up to 35 days after treatment of R. sanguineus. Similar preventive efficacy profiles for each of the Ixodid species tested suggest that CERTIFECT kills all Ixodid species starting 4h after contact as demonstrated for R. sanguineus.

Efficacy of fipronil, amitraz and (S)-methoprene combination spot-on for dogs against adult dog fleas (Ctenocephalides canis, Curtis, 1826).

A novel spot-on formulation combining fipronil, amitraz and (S)-methoprene (CERTIFECT™, Merial Limited, GA, USA) was evaluated in adult Beagle dogs in a study to determine its adulticidal efficacy against the dog flea (Ctenocephalides canis, Curtis, 1826). Sixteen dogs were randomly allocated to treatment groups: 8 dogs served as untreated controls, and 8 dogs were treated once. Treatment consisted of applying a new combination formulation to deliver at least 6.7mg fipronil/kg body weight (bw), 8.0mg amitraz/kg bw, and 6.0mg (S)-methoprene/kg bw. The combination was designed to enhance the efficacy against ticks of the original fipronil/(S)-methoprene combination. Each dog was infested with 100 adult unfed dog fleas within 24h prior to treatment and then at weekly intervals for 8 weeks after treatment. At 24h after treatment or after each subsequent infestation, each dog was combed thoroughly to remove live fleas to be counted. A single treatment with CERTIFECT provided excellent knock-down of fleas within 24h after treatment and controlled re-infestations for up to 7 weeks (efficacy ≥96.5%, p<0.05).