Targeted-pig trial on safety and immunogenicity of serum-derived extracellular vesicles enriched fractions obtained from Porcine Respiratory and Reproductive virus infections

The Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is the etiological agent of one of the most important swine diseases with a significant economic burden worldwide. Unfortunately, available vaccines are partially effective highlighting the need of novel approaches. Previously, antigenic viral proteins were described in serum-derived extracellular vesicles (EVs) from pigs previously infected with PRRSV. Here, a targeted-pig trial was designed to determine the safety and immunogenicity of such extracellular vesicles enriched fractions. Our results showed that immunizations with EV-enriched fractions from convalescence animals in combination with montanide is safe and free of virus as immunizations with up-to two milligrams of EV-enriched fractions did not induce clinical symptoms, adverse effects and detectable viral replication. In addition, this vaccine formulation was able to elicit specific humoral IgG immune response in vaccinated animals, albeit variably. Noticeably, sera from vaccinated animals was diagnosed negative when tested for PRRSV using a commercial ELISA test; thus, indicating that this new approach differentiates vaccinated from infected animals. Lastly, after priming animals with EV-enriched fractions from sera of convalescence animals and boosting them with synthetic viral peptides identified by mass spectrometry, a distinctive high and specific IFN-γ response was elicited. Altogether, our data strongly suggest the use of serum EV-enriched fractions as a novel vaccine strategy against PRRSV.Anti-CD9, Anti-CD63 and anti-CD81 antibodies were kindly donated by Francisco Sánchez-Madrid and Maria Yañez-Mo, Hospital de la Princesa, Madrid, Spain. The authors wish to particularly thank Glòria Abella for her collaboration in conducting the field study and to Marta Alcobé, Miriam Moron Font and Paula Crego Mendez for technical assistance. This study received support from Innovex Therapeutics S.L., Pinsos del Segre SA, Granja Casanyé, Grup de Sanejament Porci (GSP, Lleida, Spain) and the FEDER project (COMRDI16-1-0035-03). Sergio Montanter-Tarbes is an industrial doctorate awarded by the Government of Catalonia, Spain (No. 2014 DI 044). ISGlobal and IGTP are members of the CERCA Programme, Generalitat de Catalunya

Porcine Reproductive and Respiratory Syndrome Surveillance in breeding Herds and Nurseries Using Tongue Tips from Dead Animals

The detection capacity of Porcine Reproductive and Respiratory Syndrome virus (PRRSV) in tongues from dead animals in breeding herds (stillborns and piglets dying during the lactating period) and nursery farms (naturally dead animals) for PRRSV surveillance was evaluated. The samples were selected if pairs of serum and tongues were available from 2018 to 2020. Serum (pools of five) and exudate from tongues (one bag) were analyzed by PRRSV RT-PCR. The agreement between the serum sample procedure versus tongues exudate was assessed using a concordance test (Kappa statistic) at batch level. A total of 32 submissions, corresponding to 14 farms, had PRRSV diagnostic information for serum and tongues exudate. The overall agreement of batch classification as positive or negative, based on RT-PCR PRRSV results, between serum and tongue exudate of the 32 pairs was 76.9%. Cohen's Kappa was 0.55. The main discrepancy came from the presence of positive samples in tongues exudate and not in serum, suggesting that tongue exudate to monitor PRRSV seems to be more sensitive than serum. These results suggest that this sample procedure could be also used for PRRSV surveillance and monitoring

The Susceptibility Trends of Respiratory and Enteric Porcine Pathogens to Last-Resource Antimicrobials

Monitoring the antimicrobial susceptibility of last-resource antimicrobials for veterinary pathogens is urgently needed from a one-health perspective. The objective of this study was to analyze the antimicrobial susceptibility trends of Spanish porcine bacteria to quinolones, cephalosporins, and polymyxins. Isolates of Actinobacillus pleuropneumoniae, Pasteurella multocida, and Escherichia coli were isolated from sick pigs from 2019 to 2022. An antimicrobial susceptibility test was determined based on the minimal inhibitory concentration (MIC) following an internationally accepted methodology. The MIC categorization was based on distributing the range of MIC values in four categories, with category one being the most susceptible (lowest MIC value) and category four the least susceptible (highest MIC value). Moreover, clinical susceptibility (susceptible/non-susceptible) was also determined according to the CLSI and EUCAST clinical breakpoints. A logistic and multinomial logistic regression model was used to analyze the susceptibility data for dichotomized and categorized MIC data, respectively, for any pair of antimicrobial/microorganism. In general terms, the antimicrobial susceptibility of pig bacteria to these antimicrobials remained stable or increased in the last four years in Spain. In the case of A. pleuropneumoniae and quinolones, a significant temporal trend was observed where isolates from 2020 had significantly increased odds of being more susceptible than isolates from 2019. In the case of E. coli and polymyxins, a significant temporal trend was observed where isolates from 2020 and 2021 had significantly increased odds of being more susceptible than isolates from 2019 and 2020, respectively. Finally, significant odds of being less susceptible were only observed for cephalosporins and E. coli for 2020 versus 2019, stagnating for the rest of study period. These results provide sound data on critically important antimicrobials in swine medicine.info:eu-repo/semantics/publishedVersio

Near real-time monitoring of clinical events detected in swine herds in Northeastern Spain

Novel techniques of data mining and time series analyses allow the development of new methods to analyze information relating to the health status of the swine population in near real-time. A swine health monitoring system based on the reporting of clinical events detected at farm level has been in operation in Northeastern Spain since 2012. This initiative was supported by swine stakeholders and veterinary practitioners of the Catalonia, Aragon, and Navarra regions. The system aims to evidence the occurrence of endemic diseases in near real-time by gathering data from practitioners that visited swine farms in these regions. Practitioners volunteered to report data on clinical events detected during their visits using a web application. The system allowed collection, transfer and storage of data on different clinical signs, analysis, and modeling of the diverse clinical events detected, and provision of reproducible reports with updated results. The information enables the industry to quantify the occurrence of endemic diseases on swine farms, better recognize their spatiotemporal distribution, determine factors that influence their presence and take more efficient prevention and control measures at region, county, and farm level. This study assesses the functionality of this monitoring tool by evaluating the target population coverage, the spatiotemporal patterns of clinical signs and presumptive diagnoses reported by practitioners over more than 6 years, and describes the information provided by this system in near real-time. Between January 2012 and March 2018, the system achieved a coverage of 33 of the 62 existing counties in the three study regions. Twenty-five percent of the target swine population farms reported one or more clinical events to the system. During the study period 10,654 clinical events comprising 14,971 clinical signs from 1,693 farms were reported. The most frequent clinical signs detected in these farms were respiratory, followed by digestive, neurological, locomotor, reproductive, and dermatological signs. Respiratory disorders were mainly associated with microorganisms of the porcine respiratory disease complex. Digestive signs were mainly related to colibacilosis and clostridiosis, neurological signs to Glässer's disease and streptococcosis, reproductive signs to PRRS, locomotor to streptococcosis and Glässer's disease, and dermatological signs to exudative epidermitis.info:eu-repo/semantics/publishedVersio

Antimicrobial Susceptibility Testing of Porcine Bacterial Pathogens: Investigating the prospect of testing a representative drug for each antimicrobial family

Antimicrobial susceptibility testing is necessary to carry out antimicrobial stewardship but a limited number of drugs belonging to each antimicrobial family has to be tested for techni- cal limitations and economic resources. In this study, we have determined the minimal inhibitory concentration, using microdilution following international standards (CLSI), for 490 Actinobacillus pleuropneumoniae, 285 Pasteurella multocida, 73 Bordetella bronchiseptica, 398 Streptococcus suis and 1571 Escherichia coli strains from clinical cases collected in Spain between 2018 and 2020. The an- timicrobial susceptibility pattern was deciphered using a principal component analysis for each bacterium and a matrix correlation (high > 0.8, medium 0.5-0.8 and low < 0.5) was obtained for each pair of antimicrobials. No significant associations were observed between MIC patterns for different antimicrobial families, suggesting that co-selection mechanisms are not generally present in these porcine pathogens. However, a high correlation was observed between the fluroquinolones (mar- bofloxacin and enrofloxacin) for all mentioned pathogens and for ceftiofur and cefquinome for E. coli and S. suis. Moreover, a significant association was also observed for tetracyclines (doxycycline and oxytetracycline) and B. bronchiseptica and tildipirosin/tulathromycin for P. multocida. These results suggest that generally, a representative drug per antimicrobial class cannot be selected, however, for some drug-bug combinations, MIC values from one representative drug could be extrapolated to the whole antimicrobial family

Antimicrobial susceptibility pattern of porcine respiratory bacteria in Spain

The monitoring of antimicrobial susceptibility of pig pathogens is critical to optimize antimicrobial treatments and prevent development of resistance with a one-health approach. The aim of this study was to investigate the antimicrobial susceptibility patterns of swine respiratory pathogens in Spain from 2017 to 2019. Bacterial isolation and identification were carried out following standardized methods from samples coming from sacrificed or recently deceased pigs with acute clinical signs compatible with respiratory tract infections. Minimum inhibitory concentration (MIC) values were determined using the broth microdilution method containing a total of 10 and 7-8 antimicrobials/concentrations respectively, in accordance with the recommendations presented by the Clinical and Laboratory Standards Institute (CLSI). The obtained antimicrobial susceptibility varies between pig respiratory pathogens. Actinobacillus pleuropneumoniae (APP) and Pasteurella multocida (PM) were highly susceptible (≥90%) to ceftiofur, florfenicol and macrolides (tilmicosin, tildipirosin and tulathromycin). However, the antimicrobial susceptibility was intermediate (>60% but <90%) for amoxicillin and enrofloxacin in the case of APP and sulfamethoxazole/trimethropim and tiamulin in the case of PM. Both bacteria showed low (<60%) antimicrobial susceptibility to doxycycline. Finally, Bordetella bronchiseptica was highly susceptible only to tildipirosin and tulathromycin (100%) and its susceptibility for florfenicol was close to 50% and <30% for the rest of the antimicrobial families tested. These results emphasize the need of determining antimicrobial susceptibility in pig respiratory cases in order to optimize the antimicrobial treatment in a case-by-case scenario

Antimicrobial Susceptibility Pattern of Porcine Respiratory Bacteria in Spain

[EN] The monitoring of antimicrobial susceptibility of pig pathogens is critical to optimize antimicrobial treatments and prevent development of resistance with a one-health approach. The aim of this study was to investigate the antimicrobial susceptibility patterns of swine respiratory pathogens in Spain from 2017 to 2019. Bacterial isolation and identification were carried out following standardized methods from samples coming from sacrificed or recently deceased pigs with acute clinical signs compatible with respiratory tract infections. Minimum inhibitory concentration (MIC) values were determined using the broth microdilution method containing a total of 10 and 7–8 antimicrobials/concentrations respectively, in accordance with the recommendations presented by the Clinical and Laboratory Standards Institute (CLSI). The obtained antimicrobial susceptibility varies between pig respiratory pathogens. Actinobacillus pleuropneumoniae (APP) and Pasteurella multocida (PM) were highly susceptible (≥90%) to ceftiofur, florfenicol and macrolides (tilmicosin, tildipirosin and tulathromycin). However, the antimicrobial susceptibility was intermediate (>60% but <90%) for amoxicillin and enrofloxacin in the case of APP and sulfamethoxazole/trimethropim and tiamulin in the case of PM. Both bacteria showed low (<60%) antimicrobial susceptibility to doxycycline. Finally, Bordetella bronchiseptica was highly susceptible only to tildipirosin and tulathromycin (100%) and its susceptibility for florfenicol was close to 50% and <30% for the rest of the antimicrobial families tested. These results emphasize the need of determining antimicrobial susceptibility in pig respiratory cases in order to optimize the antimicrobial treatment in a case-by-case scenarioSIThis research received no external fundin

Near real-time monitoring of clinical events detected in swine herds in Northeastern Spain

Novel techniques of data mining and time series analyses allow the development of new methods to analyze information relating to the health status of the swine population in near real-time. A swine health monitoring system based on the reporting of clinical events detected at farm level has been in operation in Northeastern Spain since 2012. This initiative was supported by swine stakeholders and veterinary practitioners of the Catalonia, Aragon, and Navarra regions. The system aims to evidence the occurrence of endemic diseases in near real-time by gathering data from practitioners that visited swine farms in these regions. Practitioners volunteered to report data on clinical events detected during their visits using a web application. The system allowed collection, transfer and storage of data on different clinical signs, analysis, and modeling of the diverse clinical events detected, and provision of reproducible reports with updated results. The information enables the industry to quantify the occurrence of endemic diseases on swine farms, better recognize their spatiotemporal distribution, determine factors that influence their presence and take more efficient prevention and control measures at region, county, and farm level. This study assesses the functionality of this monitoring tool by evaluating the target population coverage, the spatiotemporal patterns of clinical signs and presumptive diagnoses reported by practitioners over more than 6 years, and describes the information provided by this system in near real-time. Between January 2012 and March 2018, the system achieved a coverage of 33 of the 62 existing counties in the three study regions. Twenty-five percent of the target swine population farms reported one or more clinical events to the system. During the study period 10,654 clinical events comprising 14,971 clinical signs from 1,693 farms were reported. The most frequent clinical signs detected in these farms were respiratory, followed by digestive, neurological, locomotor, reproductive, and dermatological signs. Respiratory disorders were mainly associated with microorganisms of the porcine respiratory disease complex. Digestive signs were mainly related to colibacilosis and clostridiosis, neurological signs to Glässer's disease and streptococcosis, reproductive signs to PRRS, locomotor to streptococcosis and Glässer's disease, and dermatological signs to exudative epidermitis