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.

A retrospective cohort study of 116 British pig farms was undertaken to investigate the epidemiological risk factors associated with herd breakdowns with postweaning multisystemic wasting syndrome (PMWS). Farmers reported the PMWS status of their herd (case definition 1) and, where applicable, when the disease was first suspected and what they observed; they described a prolonged increase in mortality in six to 16-week-old pigs that was not attributable to any disease known to be on their farm. There was over 90 per cent agreement on the farmers' PMWS status between the farmers and their veterinarians. Approximately 70 per cent of the breakdowns were confirmed at the laboratory (case definition 2) except during the outbreak of foot-and-mouth disease (FMD) in 2001 when it was reduced to 30 per cent. Porcine circovirus type 2 antigen was detected in pigs examined postmortem (case definition 3) in approximately 90 per cent of the farms with increased mortality. The breakdowns occurred initially in the south of England and spread west and north, as well as locally in a radial pattern from the affected farms, and there was strong statistical evidence that there was non-random space-time clustering. The risk of herd breakdowns with PMWS was not constant; therefore, for each case definition, three survival models were developed with outcome variable time to breakdown of between January 2000 and January 2001, February 2001 to September 2001 (during FMD) or October 2001 to December 2003. Exposures with a bivariable significance of P<0.20 were tested in three multivariable Cox proportional hazard models. From January 2000 to January 2001 the risk of a herd breakdown with PMWS for definitions 1, 2 and 3 was greater for farms with 600 or more breeding sows, and for definitions 1 and 3 there was an increased risk associated with the purchase of replacement gilts rather than using homebred replacements. For definitions 1 and 3 the farms where the nearest pig farm had no breeding pigs were at greater risk of a breakdown than those where the nearest farm had breeding stock, as were the farms where visitors were not requested to avoid pigs for more than three days before visiting the farm during the FMD outbreak. From October 2001, the associated risks were identical for all three case definitions; farms were at greater risk when they had 600 or more breeding sows, if visitors had not avoided contact with pigs for more than three days before visiting the farm, and when there was a farm with PMWS less than five miles away. The affected farms were more likely to have disease associated with porcine parvovirus, porcine reproduction and respiratory syndrome virus, erysipelas, Escherichia coli and salmonella. These exposures were positively associated with large herds and the farm being close to other pig farms, but did not remain in the final models for breakdown with PMWS, indicating that such farms may be at greater risk of many infectious diseases.

Porcine reproductive and respiratory syndrome (PRRS) causes decreased reproductive performance and respiratory problems in pigs. The goals of the current study were 1) to examine whether individual variation applies to infection with PRRSV European strains and 2) to investigate the association of a single nucleotide polymorphism (SNP) WUR10000125 (WUR) at the interferon-inducible guanylate-binding protein 1 gene (GBP1) with average daily gain (ADG) in PRRSV infected and uninfected pigs. The experimental procedure consisted of two trials in which pigs from negative PRRSV farms were infected with a wild-type (n=80) or vaccinated with an attenuated European PRRS virus strain (n=40) and then monitored after infection or vaccination. Viral load and ADG were determined for each pig. In a third trial, the ADG for PRRSV-free pigs was monitored. All pigs were genotyped for WUR at the GBP1 gene (AA and AG genotype were defined). Results indicated that there was individual variation in the viral load from pigs challenged with a wild-type or low virulent European PRRSV strain. Secondly, our data showed that WUR SNP was associated to ADG in vaccinated pigs. Thus, ADG in AG pigs was significantly higher than in AA ones after vaccinating with an attenuated PRRSV strain. However, the reverse happened in a PRRSV-free environment where the AA pigs were those that grew faster. Based on these results, there is a scope for selecting pigs according to their responses to PRRS virus infection with European strains and that WUR SNP may play a role in causing PRRSV resistance.

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.

The identification of resilient sows can improve reproductive performance in farms exposed to multiple challenges. A common challenge is the porcine reproductive and respiratory syndrome virus (PRRSV). A key issue to deal with disease resilience is to set up a feasible phenotyping strategy. Our aim was to develop a phenotyping criterion to discriminate susceptible from resilient sows in PRRSV-infected farms. A total of 517 Landrace x Large White gilts were classified as resilient (R) or susceptible (S) to PRRSV virus, following vaccination with MLV-PRRSV at 6 to 7 wk of age, in a PRRSV negative multiplication farm. Female piglets were phenotyped as R if their serum was negative to PRRSV at 7 and 21 d postvaccination (DPV) or as S if their serum was positive at 7 and/or 21 DPV. Amongst them, 382 gilts were transferred to a PRRSV-positive production farm, where the number of piglets born alive (NBA), stillborn (NSB), mummified (NMU), lost (NLP = NSB + NMU), and total born (NTB = NBA + NLP) were recorded for almost 3 yr. Data were collected during 2 periods according to the PRRSV farm health status, which were confirmed as either PRRSV-positive stable (endemic) or inestable (epidemic). Analyses were carried out under a Bayesian approach. The heritability for the resilience criterion was estimated using a threshold model. A linear (for NTB and NBA) and a binomial model (for NSB, NMU, and NLP) on the resilience criterion by the farm health status were used to assess the difference between R and S sows. The heritability of the resilience criterion was 0.46 (SD 0.06). The probability of a piglet being lost was greater (≥0.97) in S than in R litters, regardless of whether the delivery occurred during a PRRSV outbreak (20.5% vs. 17.0%) or not (15.8% vs. 13.7%). The lower piglet mortality rate in R sows was due to NSB, in the endemic phase (13.0% vs. 15.0% of NTB, with a posterior probability of 98% of S sows showing higher NSB than R sows), and to NMU, in the epidemic phase (4.0% vs. 8.4% of NTB, with a posterior probability of >99% of S sows showing higher NMU than R sows). During a PRRSV outbreak, the S sows were twice as likely to give birth to a mummified piglet when compared with R sows. These findings provide evidence that the described phenotyping scheme has a potential use as a PRRSV resilience criterion.

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 aim of this study was to set up antimicrobial stewardship for swine respiratory pathogens following the recommendation from the European Medicine Agency. The obtained antimicrobial susceptibility pattern recommended using antimicrobial stewardship for each clinical case instead of treatment guidelines focused on pathogens. Thus, the bacteria are isolated and the MIC values, the clinical interpretation for each antimicrobial (susceptible or resistant), additional information about the distance between the MIC obtained and the clinical breakpoint, and set up for each drug, are represented in the report provided for veterinarians. A graph from green (susceptible) to red (resistant) is enclosed for each antimicrobial and microorganism in the report. The greener, the more susceptible is the strain, and the redder, the less susceptible is the strain for each particular antimicrobial. This information could help veterinarians to select the most suitable antimicrobial from first, second, or last option antimicrobials. Thus, veterinarians should choose the antimicrobial, inside each option, with the best antimicrobial susceptibility pattern that corresponds with the greener status in the report. The information provided in the report could be useful for all clinical cases, caused by a certain bacterium within the same pig production system, if an epidemiological link could be established.

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 technical 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 antimicrobial 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 (marbofloxacin 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.