Bayesian estimation of the sensitivity and specificity of coprological and serological diagnostic tests for the detection of Ascaris suum infection on pig farms.

Coprological and serological diagnostic tests were compared to define the status of a pig farm with regard to Ascaris suum. On each of the 100 farms in France visited for the study, 10 blood samples were taken from pigs at the end of fattening (at least 22 weeks old) and 20 to 30 faecal samples were taken, depending on the category of animals present on the farm (10 sows, 10 piglets aged 10 to 12 weeks and 10 pigs at the end of fattening, aged at least 22 weeks). A SERASCA® ELISA test (Laboratory of Parasitology, Ghent University) was performed on each blood sample (cut-off 0.5) and a coprological analysis on each faecal sample. A Bayesian approach was used to estimate the sensitivity and specificity of the coprological and serological tests. A farm was considered positive if at least one A. suum egg was observed in the faecal samples. With regard to the serological test, various hypotheses were tested in order to define the number of seropositive animals required to consider a farm positive for A. suum. The coprological test has very good specificity in the search for A. suum, whether 20 or 30 samples are taken per farm. However, even with an increase in the number of samples, the sensitivity of this diagnostic approach is very low (less than 30%). On the other hand, the serological diagnostic method, which consists of taking blood samples from 10 animals at the end of fattening, has good sensitivity and seems better suited to defining the status of a farm with regard to A. suum, provided that a farm is considered seropositive only if two out of 10 samples are positive.

LSD1 controls a nuclear checkpoint in Wnt/ß-Catenin signaling to regulate muscle stem cell self-renewal.

The Wnt/ß-Catenin pathway plays a key role in cell fate determination during development and in adult tissue regeneration by stem cells. These processes involve profound gene expression and epigenome remodeling and linking Wnt/ß-Catenin signaling to chromatin modifications has been a challenge over the past decades. Functional studies of the lysine demethylase LSD1/KDM1A converge to indicate that this epigenetic regulator is a key regulator of cell fate, although the extracellular cues controlling LSD1 action remain largely unknown. Here we show that ß-Catenin is a substrate of LSD1. Demethylation by LSD1 prevents ß-Catenin degradation thereby maintaining its nuclear levels. Consistently, in absence of LSD1, ß-Catenin transcriptional activity is reduced in both MuSCs and ESCs. Moreover, inactivation of LSD1 in mouse muscle stem cells and embryonic stem cells shows that LSD1 promotes mitotic spindle orientation via ß-Catenin protein stabilization. Altogether, by inscribing LSD1 and ß-Catenin in the same molecular cascade linking extracellular factors to gene expression, our results provide a mechanistic explanation to the similarity of action of canonical Wnt/ß-Catenin signaling and LSD1 on stem cell fate.

Effect of vaccination route (intradermal vs. intramuscular) against porcine reproductive and respiratory syndrome using a modified live vaccine on systemic and mucosal immune response and virus transmission in pigs.

BACKGROUND: Porcine reproductive and respiratory syndrome (PRRS) is a viral disease with worldwide distribution and an enormous economic impact. To control PRRS virus (PRRSV) infection, modified live vaccines (MLVs) are widely used in the field, mainly administered via an intramuscular (IM) route. Currently, some MLVs are authorized for intradermal (ID) administration, which has many practical and welfare advantages. The objectives of the study were to compare the immune responses (systemic in blood and mucosal in lungs) and vaccine efficacy in preventing challenge strain transmission after IM or needle-free ID immunization of piglets with an MLV against PRRSV-1 (MLV1). METHODS: Groups of sixteen 5-week-old specific pathogen-free piglets were vaccinated with Porcilis PRRS® (MSD) either by an IM (V+ IM) or ID route (V+ ID) using an IDAL®3G device or kept unvaccinated (V-). Four weeks after vaccination, in each group, 8 out of the 16 piglets were challenged intranasally with a PRRSV-1 field strain, and one day later, the inoculated pigs were mingled by direct contact with the remaining 8 sentinel noninoculated pigs to evaluate PRRSV transmission. Thus, after the challenge, each group (V+ IM, V+ ID or V-) included 8 inoculated and 8 contact piglets. During the postvaccination and postchallenge phases, PRRSV replication (RT-PCR), PRRSV-specific antibodies (ELISA IgG and IgA, virus neutralization tests) and cell-mediated immunity (ELISPOT Interferon gamma) were monitored in blood and bronchoalveolar lavages (BALs). RESULTS: Postvaccination, vaccine viremia was lower in V+ ID pigs than in V+ IM pigs, whereas the cell-mediated immune response was detected earlier in the V+ ID group at 2 weeks postvaccination. In the BAL fluid, a very low mucosal immune response (humoral and cellular) was detected. Postchallenge, the vaccine efficacy was similar in inoculated animals with partial control of PRRSV viremia in V+ ID and V+ IM animals. In vaccinated sentinel pigs, vaccination drastically reduced PRRSV transmission with similar estimated transmission rates and latency durations for the V+ IM and V+ ID groups. CONCLUSIONS: Our results show that the tested MLV1 induced a faster cell-mediated immune response after ID immunization two weeks after vaccination but was equally efficacious after IM or ID immunization towards a challenge four weeks later. Considering the practical and welfare benefits of ID vaccination, these data further support the use of this route for PRRS MLVs.

Application of Machine Learning Prediction of Individual SARS-CoV-2 Vaccination and Infection Status to the French Serosurveillance Survey From March 2020 to 2022: Cross-Sectional Study.

BACKGROUND: The seroprevalence of SARS-CoV-2 infection in the French population was estimated with a representative, repeated cross-sectional survey based on residual sera from routine blood testing. These data contained no information on infection or vaccination status, thus limiting the ability to detail changes observed in the immunity level of the population over time. OBJECTIVE: Our aim is to predict the infected or vaccinated status of individuals in the French serosurveillance survey based only on the results of serological assays. Reference data on longitudinal serological profiles of seronegative, infected, and vaccinated individuals from another French cohort were used to build the predictive model. METHODS: A model of individual vaccination or infection status with respect to SARS-CoV-2 obtained from a machine learning procedure was proposed based on 3 complementary serological assays. This model was applied to the French nationwide serosurveillance survey from March 2020 to March 2022 to estimate the proportions of the population that were negative, infected, vaccinated, or infected and vaccinated. RESULTS: From February 2021 to March 2022, the estimated percentage of infected and unvaccinated individuals in France increased from 7.5% to 16.8%. During this period, the estimated percentage increased from 3.6% to 45.2% for vaccinated and uninfected individuals and from 2.1% to 29.1% for vaccinated and infected individuals. The decrease in the seronegative population can be largely attributed to vaccination. CONCLUSIONS: Combining results from the serosurveillance survey with more complete data from another longitudinal cohort completes the information retrieved from serosurveillance while keeping its protocol simple and easy to implement.

Mathematical modeling at the livestock-wildlife interface: scoping review of drivers of disease transmission between species.

Modeling of infectious diseases at the livestock-wildlife interface is a unique subset of mathematical modeling with many innate challenges. To ascertain the characteristics of the models used in these scenarios, a scoping review of the scientific literature was conducted. Fifty-six studies qualified for inclusion. Only 14 diseases at this interface have benefited from the utility of mathematical modeling, despite a far greater number of shared diseases. The most represented species combinations were cattle and badgers (for bovine tuberculosis, 14), and pigs and wild boar [for African (8) and classical (3) swine fever, and foot-and-mouth and disease (1)]. Assessing control strategies was the overwhelming primary research objective (27), with most studies examining control strategies applied to wildlife hosts and the effect on domestic hosts (10) or both wild and domestic hosts (5). In spatially-explicit models, while livestock species can often be represented through explicit and identifiable location data (such as farm, herd, or pasture locations), wildlife locations are often inferred using habitat suitability as a proxy. Though there are innate assumptions that may not be fully accurate when using habitat suitability to represent wildlife presence, especially for wildlife the parsimony principle plays a large role in modeling diseases at this interface, where parameters are difficult to document or require a high level of data for inference. Explaining observed transmission dynamics was another common model objective, though the relative contribution of involved species to epizootic propagation was only ascertained in a few models. More direct evidence of disease spill-over, as can be obtained through genomic approaches based on pathogen sequences, could be a useful complement to further inform such modeling. As computational and programmatic capabilities advance, the resolution of the models and data used in these models will likely be able to increase as well, with a potential goal being the linking of modern complex ecological models with the depth of dynamics responsible for pathogen transmission. Controlling diseases at this interface is a critical step toward improving both livestock and wildlife health, and mechanistic models are becoming increasingly used to explore the strategies needed to confront these diseases.

Bioengineering a miniaturized in vitro 3D myotube contraction monitoring chip to model muscular dystrophies.

Quantification of skeletal muscle functional contraction is essential to assess the outcomes of therapeutic procedures for neuromuscular disorders. Muscle three-dimensional "Organ-on-chip" models usually require a substantial amount of biological material, which rarely can be obtained from patient biopsies. Here, we developed a miniaturized 3D myotube culture chip with contraction monitoring capacity at the single cell level. Optimized micropatterned substrate design enabled to obtain high culture yields in tightly controlled microenvironments, with myotubes derived from primary human myoblasts displaying spontaneous contractions. Analysis of nuclear morphology confirmed similar myonuclei structure between obtained myotubes and in vivo myofibers, as compared to 2D monolayers. LMNA-related Congenital Muscular Dystrophy (L-CMD) was modeled with successful development of diseased 3D myotubes displaying reduced contraction. The miniaturized myotube technology can thus be used to study contraction characteristics and evaluate how diseases affect muscle organization and force generation. Importantly, it requires significantly fewer starting materials than current systems, which should substantially improve drug screening capability.

Effects of habitat fragmentation and hunting activities on African swine fever dynamics among wild boar populations.

African Swine Fever (ASF) has been slowly but steadily increasing its endemic range throughout Europe, posing an imminent risk to the pig industry. ASF transmission among wild boar occurs mainly through wild boar population movements, hence wild boar presence and density are important risk factors for introducing, maintaining, and spreading the disease. The understanding of wild boar population dynamics and their role in ASF transmission and persistence remains limited. It is crucial to gain knowledge in this area to improve wildlife management while minimizing the risks for ASF introduction and spread. We adapted an individual-based spatio-temporal stochastic model developed by Halasa et al. (2019) and tailored it to two regions in France. The model assessed yearly hunting activity, the carcass persistence seasonality, and the specific landscape characteristics of the Franco-Belgian border region and the Pyrénées-Atlantiques department. Following the establishment of local population dynamics through preliminary runs of the model, the model was run 100 iterations over 8 years in the two study areas where ASF was randomly seeded after the 2nd year of simulation. For each scenario, the model was initiated with 500 wild boar groups randomly spread across the study areas. Hunting activities were included and excluded to assess the impact on population growth and ASF spread. Results showed an ever-growing wild boar population for all scenarios, which was balanced when hunting activities were included. When introducing ASF, the wild boar populations were dramatically impacted in both areas with a decrease of 63 % of the population at the Franco-Belgian border and 86 % in the Pyrénées-Atlantiques department. Habitat fragmentation and landscape connectivity were highlighted as important factors shaping ASF propagation. The Franco-Belgian border, which had the most fragmented habitat with unsuitable areas for wild boars, was shown to limit wild boar movements, reducing the probability, and spread of ASF across the landscape. The lack of connectivity was reflected in a less effective transmission and lower number of infected groups (406 versus 467). In contrast, the epidemic duration was lengthened in the fragmented habitat compared to the homogenous area (2.6 years vs 1.6 years). This study provided information on defining and implementing control measures in case of an ASF incursion, since delimitation of the area via fences artificially induces landscape fragmentation, which is important for controlling ASF outbreaks.

DESCRIPTIVE EPIDEMIOLOGY OF THE MAIN INTERNAL PARASITES ON ALTERNATIVE PIG FARMS IN FRANCE.

Alternative pig farms, which do not raise animals in closed buildings with slatted and/or concrete floors, have critical points that need particular attention. Internal parasitism is one, as the farming conditions in such structures are more favorable to the development and survival of parasites. The objectives of this study, carried out on 70 alternative farms in continental France, were to (i) estimate the frequency and level of infestation by the main internal parasites on these farms, and (ii) define their typology according to the level of parasitism. For this purpose, fecal samples were taken for coprological analysis from 10 sows, 10 pigs aged 10-12 wk, and 10 pigs at the end of the fattening period. Blood samples were also taken for serological analysis (targeting Ascaris suum and Toxoplasma gondii) from 10 sows and 10 pigs at the end of the fattening period. Of the 70 farms, only 5 had no helminth egg or coccidian oocyst. Coccidia oocysts were observed in 79% of the farms, while eggs of Oesophagostomum spp./Hyostrongylus rubidus, Ascaris suum, and Trichuris suis were found in 47%, 16%, and 36% of the farms, respectively. On each infested farm, an average of 56.8% of sows, 23.8% of grower pigs, and 38.9% of finisher pigs were parasitized. At least 1 Ascaris suum-seropositive finisher pig was found on 91% of the farms, and at least 1 Toxoplasma gondii-seropositive finisher pig or sow on 60% of the farms. Data on housing, animal management, and health management (particularly parasite control) were collected to characterize the typology of farms according to their level of parasitism. The variables defining these farm typologies differed according to the parasites. Access to the outdoors for breeding stock was a characteristic of the farms most heavily infested with helminths or T. gondii. Conversely, the farms with the lowest frequency of coccidia oocyst infestation were characterized by free-range farrowing facilities and also by the presence of slatted floors, mostly plastic in our study, rather than straw bedding in the farrowing rooms. The level of biosecurity concerning the storage of straw for pig bedding was another discriminating factor for parasitism level of helminths and T. gondii. Farms with the highest levels of helminth parasitism were more likely not to practice an all-in-all-out postweaning system and to deworm their grower/finisher pigs less frequently than farms with the lowest levels of helminth parasitism.

Complex network analysis to understand trading partnership in French swine production.

The circulation of livestock pathogens in the pig industry is strongly related to animal movements. Epidemiological models developed to understand the circulation of pathogens within the industry should include the probability of transmission via between-farm contacts. The pig industry presents a structured network in time and space, whose composition changes over time. Therefore, to improve the predictive capabilities of epidemiological models, it is important to identify the drivers of farmers' choices in terms of trade partnerships. Combining complex network analysis approaches and exponential random graph models, this study aims to analyze patterns of the swine industry network and identify key factors responsible for between-farm contacts at the French scale. The analysis confirms the topological stability of the network over time while highlighting the important roles of companies, types of farm, farm sizes, outdoor housing systems and batch-rearing systems. Both approaches revealed to be complementary and very effective to understand the drivers of the network. Results of this study are promising for future developments of epidemiological models for livestock diseases. This study is part of the One Health European Joint Programme: BIOPIGEE.

Modelling African swine fever virus spread in pigs using time-respective network data: Scientific support for decision makers.

African swine fever (ASF) represents the main threat to swine production, with heavy economic consequences for both farmers and the food industry. The spread of the virus that causes ASF through Europe raises the issues of identifying transmission routes and assessing their relative contributions in order to provide insights to stakeholders for adapted surveillance and control measures. A simulation model was developed to assess ASF spread over the commercial swine network in France. The model was designed from raw movement data and actual farm characteristics. A metapopulation approach was used, with transmission processes at the herd level potentially leading to external spread to epidemiologically connected herds. Three transmission routes were considered: local transmission (e.g. fomites, material exchange), movement of animals from infected to susceptible sites, and transit of trucks without physical animal exchange. Surveillance was represented by prevalence and mortality detection thresholds at herd level, which triggered control measures through movement ban for detected herds and epidemiologically related herds. The time from infection to detection varied between 8 and 21 days, depending on the detection criteria, but was also dependent on the types of herds in which the infection was introduced. Movement restrictions effectively reduced the transmission between herds, but local transmission was nevertheless observed in higher proportions highlighting the need of global awareness of all actors of the swine industry to mitigate the risk of local spread. Raw movement data were directly used to build a dynamic network on a realistic timescale. This approach allows for a rapid update of input data without any pre-treatment, which could be important in terms of responsiveness, should an introduction occur.

Do Rubber Floor Mats Prevent Lameness in Gestating Sows Housed in Large Groups? A Field Experiment on Three Commercial Farms in France.

Lameness and foot disorders are major health and welfare issues in intensive swine production systems. They are exacerbated when sows are housed in large groups on slatted concrete floors during gestation. Our study aimed to assess the effect of rubber mats in the lying area of the gestation pen on lameness and leg health in gestating sows housed in large pens in commercial conditions. The study was conducted on three commercial farms over two successive gestations. A total of 582 Large White × Landrace sows, housed in 10 static groups, were enrolled: 5 groups in pens with rubber mats and 5 groups on slatted concrete floors. Lameness, bursitis, leg injuries, claw growth defects and claw lesions were measured at the beginning, middle and end of each gestation period. The rubber mats decreased the risk of suffering from bursitis, but had no effect on the risk of lameness, leg injuries, claw growth defects or claw lesions. Sows housed on rubber mats were heavily soiled compared with those on slatted concrete floors because the mats were not perforated for slurry evacuation. Locomotion disorders and foot lesions remained prevalent despite the rubber mats in the lying area of the gestation pens, but adding rubber mats in service rooms and farrowing crates may produce better results.

Phenotypic and Genetic Evolutions of a Porcine Reproductive and Respiratory Syndrome Modified Live Vaccine after Limited Passages in Pigs.

Modified live vaccines (MLVs) against the porcine reproductive and respiratory syndrome virus (PRRSV) have been regularly associated with safety issues, such as reversion to virulence. In order to characterize the phenotypic and genetic evolution of the PRRSV-1 DV strain from the Porcilis® PRRS MLV after limited passages in pigs, three in vivo experiments were performed. Trial#1 aimed (i) at studying transmission of the vaccine strain from vaccinated to unvaccinated contact pigs. Trial#2 and Trial#3 were designed (ii) to assess the reproducibility of Trial#1, using another vaccine batch, and (iii) to compare the virulence levels of two DV strains isolated from vaccinated (passage one) and diseased contact pigs (passage two) from Trial#1. DV strain isolates from vaccinated and contact pigs from Trial#1 and Trial#2 were submitted to Next-Generation Sequencing (NGS) full-genome sequencing. All contact animals from Trial#1 were infected and showed significantly increased viremia compared to vaccinated pigs, whereas no such change was observed during Trial#2. In Trial#3, viremia and transmission were higher for inoculated pigs with passage two of the DV strain, compared with passage one. In this study, we showed that the re-adaptation of the DV strain to pigs is associated with faster replication and increased transmission of the vaccine strain. Punctually, a decrease of attenuation of the DV vaccine strain associated with clinical signs and increased viremia may occur after limited passages in pigs. Furthermore, we identified three mutations linked to pig re-adaptation and five other mutations as potential virulence determinants.

Mechanistic modelling of African swine fever: A systematic review.

The spread of African swine fever (ASF) poses a grave threat to the global swine industry. Without an available vaccine, understanding transmission dynamics is essential for designing effective prevention, surveillance, and intervention strategies. These dynamics can often be unraveled through mechanistic modelling. To examine the assumptions on transmission and objectives of the mechanistic models of ASF, a systematic review of the scientific literature was conducted. Articles were examined across multiple epidemiological and model characteristics, with filiation between models determined through the creation of a neighbor-joined tree using phylogenetic software. Thirty-four articles qualified for inclusion, with four main modelling objectives identified: estimating transmission parameters (11 studies), assessing determinants of transmission (7), examining consequences of hypothetical outbreaks (5), assessing alternative control strategies (11). Population-based (17), metapopulation (5), and individual-based (12) model frameworks were represented, with population-based and metapopulation models predominantly used among domestic pigs, and individual-based models predominantly represented among wild boar. The majority of models (25) were parameterized to the genotype II isolates currently circulating in Europe and Asia. Estimated transmission parameters varied widely among ASFV strains, locations, and transmission scale. Similarly, parameter assumptions between models varied extensively. Uncertainties on epidemiological and ecological parameters were usually accounted for to assess the impact of parameter values on the modelled infection trajectory. To date, almost all models are host specific, being developed for either domestic pigs or wild boar despite the fact that spillover events between domestic pigs and wild boar are evidenced to play an important role in ASF outbreaks. Consequently, the development of more models incorporating such transmission routes is crucial. A variety of codified and hypothetical control strategies were compared however they were all a priori defined interventions. Future models, built to identify the optimal contributions across many control methods for achieving specific outcomes should provide more useful information for policy-makers. Further, control strategies were examined in competition with each other, which is opposed to how they would actually be synergistically implemented. While comparing strategies is beneficial for identifying a rank-order efficacy of control methods, this structure does not necessarily determine the most effective combination of all available strategies. In order for ASFV models to effectively support decision-making in controlling ASFV globally, these modelling limitations need to be addressed.

Compressed Brown Algae as a Potential Environmental Enrichment Material in Growing Pigs.

In barren housing conditions, enrichment materials are given to pigs to improve their welfare. Here, we assessed the suitability of an algae-based cylinder as an enrichment material on the behavioral, physiological, health and productivity welfare indicators of pigs. Algae was compared with metal chains and wood logs. The study involved 444 pigs from two successive batches on one single farm. During the suckling period, half of the pigs received algae and the control pigs received no material. After weaning and until the end of fattening, algae, wood or chains were equally distributed among the pigs. Consumption of algae cylinders was different between pens and between batches. After weaning and during the fattening period, although the results differed between batches, no significant difference was observed in the object manipulations. Salivary cortisol, used as biomarker to measure the stress levels after pig transfers, were similar between the treatments. Enrichment material made from algae had no negative effect on pig health and no effect on performance and body condition. Regarding their characteristics and according to Commission Recommendation (EU) 2016/336 classification, algae cylinders can be categorized as suboptimal enrichment materials, although the present results suggest that it does not significantly improve pig welfare compared to a metal chain, which is categorized as a material of marginal interest.

Spatiotemporal clustering and Random Forest models to identify risk factors of African swine fever outbreak in Romania in 2018-2019.

African swine fever (ASF) has affected Romania since July 2017, with considerable economic and social consequences, despite the implementation of control measures mainly based on stamping out of infected pig populations. On the basis of the 2973 cumulative recorded cases up to September 2019 among wild boars and domestic pigs, analysis of the epidemiological characteristics could help to identify the factors favoring the persistence and spread of ASF. A statistical framework, based on a random forest methodology, was therefore developed to assess the spatiotemporal features of the epidemics and their relationships with environmental, human, and agricultural factors. The landscape of Romania was associated with the infection dynamics, particularly concerning forested and wetland areas. Waterways were also identified as a pivotal factor, raising questions about possible waterborne transmission since these waterways are often used as a water supply for backyard holdings. However, human activity was clearly identified as the main risk factor for the spread of ASF. Although the situation in Romania cannot be directly transposed to intensive pig farming countries, the findings of this study highlight the need for strict biosecurity measures on farms, and during transportation, to avoid ASF transmission at large geographic and temporal scales.

A between-herd data-driven stochastic model to explore the spatio-temporal spread of hepatitis E virus in the French pig production network.

Hepatitis E virus is a zoonotic pathogen for which pigs are recognized as the major reservoir in industrialised countries. A multiscale model was developed to assess the HEV transmission and persistence pattern in the pig production sector through an integrative approach taking into account within-farm dynamics and animal movements based on actual data. Within-farm dynamics included both demographic and epidemiological processes. Direct contact and environmental transmission routes were considered along with the possible co-infection with immunomodulating viruses (IMVs) known to modify HEV infection dynamics. Movements were limited to 3,017 herds forming the largest community on the swine commercial network in France and data from the national pig movement database were used to build the contact matrix. Between-herd transmission was modelled by coupling within-herd and network dynamics using the SimInf package. Different introduction scenarios were tested as well as a decrease in the prevalence of IMV-infected farms. After introduction of a single infected gilt, the model showed that the transmission pathway as well as the prevalence of HEV-infected pigs at slaughter age were affected by the type of the index farm, the health status of the population and the type of the infected farms. These outcomes could help design HEV control strategies at a territorial scale based on the assessment of the farms' and network's risk.

PCV2 co-infection does not impact PRRSV MLV1 safety but enhances virulence of a PRRSV MLV1-like strain in infected SPF pigs.

Co-infection by a type 1 modified live vaccine-like strain (MLV1-like) of porcine reproductive and respiratory syndrome virus (PRRSV) and a type 2 porcine circovirus (PCV2) was identified on a French pig farm with post-weaning multisystemic wasting syndrome (PMWS). An in vivo experiment was set up to characterize the virulence level of the MLV1-like strain compared with the parental MLV1 strain, and to assess the impact of PCV2 co-infection on the pathogenicity of both PRRSV strains. Six groups of six pigs each were inoculated only with either one of the two PRRSV strains or with PCV2, or co-inoculated with PCV2 and MLV1 or PCV2 and MLV1-like strains. Six contact pigs were added to each inoculated group to assess viral transmission. The animals were monitored daily for 35 days post-inoculation for clinical symptoms. Blood and nasal swabs were sampled twice a week, and tissue samples were collected during necropsy for viral quantification. Compared to MLV1-infected pigs, animals infected with the MLV1-like strain had increased PRRSV viremia and nasal shedding, a higher viral load in the tonsils, and lymph node hypertrophy at microscopic level. PCV2 co-infection did not influence clinical, virologic or transmission parameters for MLV1, but co-infected MLV1-like/PCV2 pigs had the most severe lung lesions, the highest viremia in contact animals and the highest transmission rate. Our study demonstrated that the MLV1 strain tested was safe when co-inoculated with PCV2 in piglets. However, co-infection by the MLV1-like strain and PCV2 resulted in increased virulence compared with that due to a single infection.

Tackling hepatitis E virus spread and persistence on farrow-to-finish pig farms: Insights from a stochastic individual-based multi-pathogen model.

Hepatitis E virus (HEV) is a zoonotic agent of which domestic pigs have been recognised as the main reservoir in industrialised countries. The great variability in HEV infection dynamics described on different pig farms may be related to the influence of other pathogens, and in particular viruses affecting pigs' immune response. The objective of this study was to develop a multi-pathogen modelling approach to understand the conditions under which HEV spreads and persists on a farrow-to-finish pig farm taking into account the fact that pigs may be co-infected with an intercurrent pathogen. A stochastic individual-based model was therefore designed that combines a population dynamics model, which enables us to take different batch rearing systems into account, with a multi-pathogen model representing at the same time the dynamics of both HEV and the intercurrent pathogen. Based on experimental and field data, the epidemiological parameters of the HEV model varied according to the pig's immunomodulating virus status. HEV spread and persistence was found to be very difficult to control on a farm with a 20-batch rearing system. Housing sows in smaller groups and eradicating immunomodulating pathogens would dramatically reduce the prevalence of HEV-positive livers at slaughter, which would drop from 3.3% to 1% and 0.2% respectively (p-value < 0.01). It would also decrease the probability of HEV on-farm persistence from 0.6 to 0 and 0.34 respectively (p-value < 0.01) on farms with a 7 batch rearing system. A number of farming practices, such as limiting cross-fostering, reducing the size of weaning pens and vaccinating pigs against immunomodulating viruses, were also shown to be pivotal factors for decreasing HEV spread and persistence.

Assessment of the impact of forestry and leisure activities on wild boar spatial disturbance with a potential application to ASF risk of spread.

In Europe, African swine fever virus (ASFV) is one of the most threatening infectious transboundary diseases of domestic pigs and wild boar. In September 2018, ASF was detected in wild boar in the South of Belgium. France, as a bordering country, is extremely concerned about the ASF situation in Belgium, and an active preparedness is ongoing in the country. One of the questions raised by this situation relates to disturbing activities that may affect wild boar movements and their possible impact on the spread of ASFV. Despite evidence of disturbance related to hunting practices, there is a paucity of information on the impact of forestry and human leisure activities. To assess this impact on wild boar movements, a systematic review was first conducted but very few useful data were obtained. For this reason, an expert elicitation was carried out by the French Agency for Food, Environmental and Occupational Health & Safety in order to deal with this knowledge gap. A total of 30 experts originating from France and adjacent neighbouring countries (Spain, Belgium and Switzerland) were elicited about the relative importance of six factors of spatial disturbance of wild boar (noise, smell, invasion of space, modification of the environment, duration and frequency of the activity). Then, for each factor of disturbance, they were asked about the impact of 16 different commercial forestry and human leisure activities. A global weighted score was estimated in order to capture the variability of a wide range of territorial conditions and the uncertainty of expert elicitation. This estimate permitted ranking all 16 activities and aggregating them in three groups according to their potential for disturbance of wild boar, using a regression tree analysis. The results of this expert elicitation provide a methodological approach that may be useful for French and other European decision makers and stakeholders involved in the crisis management of ASF.

Controlling hepatitis E virus in the pig production sector: Assessment of the technical and behavioural feasibility of on-farm risk mitigation strategies.

Hepatitis E virus (HEV) is a zoonotic agent with pigs as the main reservoir in industrialised countries. Recent studies conducted on pig farms, in experimental conditions or through modelling approaches, have led to a better understanding of the spread of HEV on pig farms. The findings have also made it possible to define a set of measures to reduce HEV prevalence and the risk of marketing contaminated products. The objective of this study was to assess the feasibility of a set of HEV control strategies on pig farms. Individual semi-structured interviews were conducted with farmers, veterinarians and farming advisors to collect general data, their level of knowledge of HEV, their opinion on the technical feasibility of certain changes in practices, their perception of the respective responsibilities of the different stakeholders, and their feelings about the importance of the issue, following the framework of the Theory of Planned Behaviour. The interviews made it possible to highlight potential barriers and preferred motivators for the implementation of on-farm risk mitigation strategies. Barriers included lack of knowledge, scientific gaps, perceived inability to control HEV, and low perception of the importance of the issue. Motivators included professional satisfaction, family recognition, and the opportunity to achieve higher quality standards. Three clusters of stakeholders were also identified, with a group of leaders who could help unlock reluctance and disseminate innovations. This type of behavioural approach appeared useful to help risk managers facilitate zoonotic control on pig farms.