Evaluation of Extended Storage of Swine Complete Feed for Inactivation of Viral Contamination and Effect on Nutritional, Microbiological, and Toxicological Profiles.

The extended storage of feed ingredients has been suggested as a method to mitigate the risk of pathogen transmission through contaminated ingredients. To validate the approach of extended storage of complete swine feed for the inactivation of swine viruses, an experiment was conducted wherein swine feed was inoculated with 10 mL of 1 × 105 TCID50/mL of porcine reproductive and respiratory syndrome virus (PRRSV), porcine epidemic diarrhea virus (PEDV), and Senecavirus A (SVA) and stored for 58 d at 23.9 °C. Measures of feed quality were also evaluated at the initiation and conclusion of the storage period including screening for mycotoxins, characterization of select microbiological measures, and stability of phytase and dietary vitamins. Storing feed for 58 d under either ambient or anaerobic and temperature-controlled storage conditions did not result in substantial concerns related to microbiological profiles. Upon exposure to the feed following 58 d of storage in a swine bioassay, previously confirmed naïve pigs showed no signs of PEDV or SVA replication as detected by the PCR screening of oral fluids and serum antibody screening. Infection with SVA was documented in the positive control room through diagnostic testing through the State of Minnesota. For PRRSV, the positive control room demonstrated infection. For rooms consuming inoculated feed stored for 58 d, there was no evidence of PRRSV infection with the exception of unintentional aerosol transmission via a documented biocontainment breach. In summary, storing feed for 58 d at anaerobic and temperature-controlled environmental conditions of 23.9 °C validates that the extended storage of complete swine feed can be a method to reduce risks associated with pathogen transmission through feed while having minimal effects on measures of nutritional quality.

A Molecular and Epidemiological Description of a Severe Porcine Reproductive and Respiratory Syndrome Outbreak in a Commercial Swine Production System in Russia.

Porcine reproductive and respiratory syndrome (PRRS) is an economically devastating disease of swine in many parts of the world. Porcine reproductive and respiratory syndrome virus (PRRSV) type 1 is endemic in Europe, and prevalence of the subtypes differ spatially. In this study, we investigated a severe PRRS outbreak reported in 30 farms located in eastern Russia that belong to a large swine production company in the region that was also experiencing a pseudorabies outbreak in the system. Data included 28 ORF5 sequences from samples across 18 of the 25 infected sites, reverse transcriptase real-time polymerase chain reaction (RT-qPCR) results from diagnostic testing, reports of clinical signs, and animal movement records. We observed that the outbreak was due to two distinct variants of wildtype PRRSV type 1 subtype 1 with an average genetic distance of 15%. Results suggest that the wildtype PRRSV variants were introduced into the region around 2019, before affecting this production system (i.e., sow farms, nurseries, and finisher farms). Clinical signs did not differ between the variants, but they did differ by stage of pig production. Biosecurity lapses, including movement of animals from infected farms contributed to disease spread.

Monoglyceride reduces viability of porcine epidemic diarrhoea virus in feed and prevents disease transmission to post-weaned piglets.

Outbreaks of African swine fever virus (ASFv) and porcine epidemic diarrhoea virus (PEDv) have revealed the susceptibility of livestock to disease transmitted through feed. Several viruses, including PEDv, survive in feed and may introduce disease that causes significant morbidity and mortality. In 2013, PEDv, which causes severe diarrhoea and vomiting, reached North America after spreading for decades across Eurasia. The global exchange of ingredients has created demand for products that prevent disease transmission from feed. Formaldehyde-based products are highly effective at inactivating enveloped viruses when applied at 3.25 kg/t. Alternative products to formaldehyde, including carboxylic acids, essential oils and medium chain fatty acids (MCFAs), have exhibited mixed efficacy against PEDv and require application rates higher than formaldehyde. Amphiphilic molecules like MCFAs disrupt the bilayer-lipid membranes that protect viral nucleic acids through the formation of micelles. Monoglycerides form micelles at lower concentrations than MCFAs, which suggests they may be more potent against enveloped viruses. The potential efficacy of monoglycerides against enveloped viruses in feed led to the development and examination of an experimental monoglyceride blend. The proprietary monoglyceride blend significantly (p < .0001) reduced PEDv viability in vitro after application to feed at 1.5, 2.5 and 3.5 kg/t. The monoglyceride was tested in a natural feeding behaviour challenge model in piglets. The feed was contaminated with ice-blocks containing viable PEDv, and the piglets were exposed to PEDv through the feed bin for 20 days. At the end of the 20-day challenge period, all pigs were rectally swabbed and tested for PEDv by qPCR. In the untreated control group 54.8% of the piglets tested positive for PEDv, whereas none of the MCFA-treated feed (10 kg/t inclusion) transmitted PEDv. Strikingly, the monoglyceride-treated groups (1.5, 2.5 and 3.5 kg/t) all exhibited 100% protection from PEDv. These data support the use of this proprietary monoglyceride blend in mitigation and prevention of viral disease transmission to piglets from contaminated feed.

The risk and mitigation of foot-and-mouth disease virus infection of pigs through consumption of contaminated feed.

Transboundary movement of animal feed and feed ingredients has been identified as a route for pathogen incursions. While imports of animals and animal-derived products are highly regulated for the purpose of infectious disease prevention, there has been less consideration of the viability of infectious agents in inanimate products, such as feed. This study investigated the ability of foot-and-mouth disease virus (FMDV) to remain infectious as a contaminant of commercial whole pig feed and select pig feed ingredients, and to establish the minimum infectious dose (MIDF ) required to cause foot-and-mouth disease (FMD) in pigs that consumed contaminated feed. FMDV viability in vitro varied depending on virus strain, feed product, and storage temperature, with increased duration of infectivity in soybean meal compared to pelleted whole feed. Specifically, both strains of FMDV evaluated remained viable through to the end of the 37 day observation period in experimentally contaminated soybean meal stored at 4 or 20°C . The MIDF for pigs consuming contaminated feed varied across virus strains and exposure duration in the range of 106.2 to 107 TCID50 . The ability of FMDV to cause infection in exposed pigs was mitigated by pre-treatment of feed with two commercially available feed additives, based on either formaldehyde (SalCURB®) or lactic acid (Guardian™). Our findings demonstrate that FMDV may remain infectious in pig feed ingredients for durations compatible with transoceanic transport. Although the observed MIDF was relatively high, variations in feeding conditions and biophysical characteristics of different virus strains may alter the probability of infection. These findings may be used to parameterize modelling of the risk of FMDV incursions and to regulate feed importation to minimize the risk of inadvertent importation.

Evaluation of the Impact of Antimicrobial Use Protocols in Porcine Reproductive and Respiratory Syndrome Virus-Infected Swine on Phenotypic Antimicrobial Resistance Patterns.

A longitudinal study was conducted to assess the impact of different antimicrobial exposures of nursery-phase pigs on patterns of phenotypic antimicrobial resistance (AMR) in fecal indicator organisms throughout the growing phase. Based on practical approaches used to treat moderate to severe porcine reproductive and respiratory syndrome virus (PRRSV)-associated secondary bacterial infections, two antimicrobial protocols of differing intensities of exposure [44.1 and 181.5 animal-treatment days per 1000 animal days at risk (ATD)] were compared with a control group with minimal antimicrobial exposure (2.1 ATD). Litter-matched pigs (n = 108) with no prior antimicrobial exposure were assigned randomly to the treatment groups. Pen fecal samples were collected nine times during the wean-to-finish period and cultured for Escherichia coli and Enterococcus spp. Antimicrobial-susceptibility testing was conducted using NARMS Gram-negative and Gram-positive antibiotic panels. Despite up to 65-fold difference in ATD, few and modest differences were observed between groups and over time. Resistance patterns at marketing overall remained similar to those observed at weaning, prior to any antimicrobial exposures. Those differences observed could not readily be reconciled with the patterns of antimicrobial exposure. Resistance of E. coli to streptomycin was higher in the group exposed to 44.1 ATD, but no aminoglycosides were used. In all instances where resistances differed between time points, the higher resistance occurred early in the trial prior to any antimicrobial exposures. These minimal impacts on AMR despite substantially different antimicrobial exposures point to the lack of understanding of the drivers of AMR at the population level and the likely importance of factors other than antimicrobial exposure. IMPORTANCE Despite a recognized need for more longitudinal studies to assess the effects of antimicrobial use on resistance in food animals, they remain sparse in the literature, and most longitudinal studies of pigs have been observational. The current experimental study had the advantages of greater control of potential confounding, precise measurement of antimicrobial exposures which differed markedly between groups and tracking of pigs until market age. Overall, resistance patterns were remarkably stable between the treatment groups over time, and the differences observed could not be readily reconciled with the antimicrobial exposures, indicating the likely importance of other determinants of AMR at the population level.

An assessment of enhanced biosecurity interventions and their impact on porcine reproductive and respiratory syndrome virus outbreaks within a managed group of farrow-to-wean farms, 2020-2021.

Introduction: Porcine reproductive and respiratory syndrome virus (PRRSV) has been a challenge for the U.S. swine industry for over 30 years, costing producers more than $600 million annually through reproductive disease in sows and respiratory disease in growing pigs. In this study, the impact of enhanced biosecurity practices of site location, air filtration, and feed mitigation was assessed on farrow-to-wean sites managed by a large swine production management company in the Midwest United States. Those three factors varied in the system that otherwise had implemented a stringent biosecurity protocol on farrow-to-wean sites. The routine biosecurity followed commonplace activities for farrow-to-wean sites that included but were not limited to visitor registration, transport disinfection, shower-in/shower-out procedures, and decontamination and disinfection of delivered items and were audited. Methods: Logistic regression was used to evaluate PRRSV infection by site based on the state where the site is located and air filtration use while controlling for other variables such as vaccine status, herd size, and pen vs. stall. A descriptive analysis was used to evaluate the impact of feed mitigation stratified by air filtration use. Results: Sites that used feed mitigates as additives in the diets, air filtration of barns, and that were in less swine-dense areas appeared to experience fewer outbreaks associated with PRRSV infection. Specifically, 23.1% of farms that utilized a feed mitigation program experienced PRRSV outbreaks, in contrast to 100% of those that did not. Sites that did not use air filtration had 20 times greater odds of having a PRRSV outbreak. The strongest protective effect was found when both air filtration and feed mitigation were used. Locations outside of Minnesota and Iowa had 98.5-99% lesser odds of infection as well. Discussion: Enhanced biosecurity practices may yield significant protective effects and should be considered for producers in swine-dense areas or when the site contains valuable genetics or many pigs.

Quantification of soya-based feed ingredient entry from ASFV-positive countries to the United States by ocean freight shipping and associated seaports.

African swine fever virus (ASFV) can survive in soya-based products for 30 days with T ½ ranging from 9.6 to 12.9 days in soya bean meals and soya oil cake. As the United States imports soya-based products from several ASFV-positive countries, knowledge of the type and quantity of these specific imports, and their ports of entry (POE), is necessary information to manage risk. Using the data from the International Trade Commission Harmonized Tariff Schedule website in conjunction with pivot tables, we analysed imports across air, land and sea POE of soya-based products from 43 ASFV-positive countries to the United States during 2018 and 2019. In 2018, 104,366 metric tons (MT) of soya-based products, specifically conventional and organic soya bean meal, soya beans, soya oil cake and soya oil were imported from these countries into the United States via seaports only. The two largest suppliers were China (52.7%, 55,034 MT) and the Ukraine (42.9%, 44,775 MT). In 2019, 73,331 MT entered the United States and 54.7% (40,143 MT) came from the Ukraine and 8.4% (6,182 MT) from China. Regarding POE, 80.9%-83.2% of soya-based imports from China entered the United States at the seaports of San Francisco, CA, and Seattle, WA, while 89.4%-100% entered from the Ukraine via the seaports of New Orleans, LA, and Charlotte, NC. Analysis of five-year trends (2015-2019) of the volume of soya imports from China indicated reduction over time (with a noticeably sharp decrease between 2018 and 2019), and seaport utilization was consistent. In contrast, volume remained high for Ukrainian soya imports, and seaport utilization was inconsistent. Overall, this exercise introduced a new approach to collect objective data on an important risk factor, providing researchers, government officials and industry stakeholders a means to objectively identify and quantify potential channels of foreign animal disease entry into the United States.

An evaluation of additives for mitigating the risk of virus-contaminated feed using an ice-block challenge model.

The role of animal feed as a vehicle for the transport and transmission of viral diseases was first identified during the porcine epidemic diarrhoea virus (PEDV) epidemic in North America. Since that time, various feed additives have been evaluated at the laboratory level to measure their effect on viral viability and infectivity in contaminated feed using bioassay piglet models. While a valid first step, the conditions of these studies were not representative of commercial swine production. Therefore, the purpose of this study was to evaluate the ability of feed additives to mitigate the risk of virus-contaminated feed using a model based on real-world conditions. This new model used an 'ice-block' challenge, containing equal concentrations of porcine reproductive and respiratory syndrome virus (PRRSV), Senecavirus A (SVA) and PEDV, larger populations of pigs, representative commercial facilities and environments, along with realistic volumes of complete feed supplemented with selected additives. Following supplementation, the ice block was manually dropped into designated feed bins and pigs consumed feed by natural feeding behaviour. After challenge, samples were collected at the pen level (feed troughs, oral fluids) and at the animal level (clinical signs, viral infection, growth rate, and mortality) across five independent experiments involving 15 additives. In 14 of the additives tested, pigs on supplemented diets had significantly greater average daily gain (ADG), significantly lower clinical signs and infection levels, and numerically lower mortality rates compared to non-supplemented controls. In conclusion, the majority of the additives evaluated mitigated the effects of PRRSV 174, PEDV and SVA in contaminated feed, resulting in improved health and performance.

Quantifying Individual Response to PRRSV Using Dynamic Indicators of Resilience Based on Activity.

Pigs are faced with various perturbations throughout their lives, some of which are induced by management practices, others by natural causes. Resilience is described as the ability to recover from or cope with a perturbation. Using these data, activity patterns of an individual, as well as deviations from these patterns, can potentially be used to quantify resilience. Dynamic indicators of resilience (DIORs) may measure resilience on a different dimension by calculating variation, autocorrelation and skewness of activity from the absolute activity data. The aim of this study was to investigate the potential of using DIORs of activity, such as average, root mean square error (RMSE), autocorrelation or skewness as indicators of resilience to infection with the Porcine Reproductive and Respiratory Syndrome Virus (PRRSV). For this study, individual activity was obtained from 232 pigs equipped with ear tag accelerometers and inoculated with PRRSV between seven and 9 weeks of age. Clinical scores were assigned to each individual at 13 days post-challenge and used to distinguish between a resilient and non-resilient group. Mortality post-challenge was also recorded. Average, RMSE, autocorrelation and skewness of activity were calculated for the pre- and post-challenge phases, as well as the change in activity level pre- vs. post-challenge (i.e., delta). DIORs pre-challenge were expected to predict resilience to PRRSV in the absence of PRRSV infection, whereas DIORs post-challenge and delta were expected to reflect the effect of the PRRSV challenge. None of the pre-challenge DIORs predicted morbidity or mortality post-challenge. However, a higher RMSE in the 3 days post-challenge and larger change in level and RMSE of activity from pre- to post-challenge tended to increase the probability of clinical signs at day 13 post-infection (poor resilience). A higher skewness post-challenge (tendency) and a larger change in skewness from pre- to post-challenge increased the probability of mortality. A decrease in skewness post-challenge lowered the risk of mortality. The post-challenge DIOR autocorrelation was neither linked to morbidity nor to mortality. In conclusion, results from this study showed that post-challenge DIORs of activity can be used to quantify resilience to PRRSV challenge.

The risk of viral transmission in feed: What do we know, what do we do?

The role of animal feed as a vehicle for the transport and transmission of viral diseases was first identified in 2014 during the porcine epidemic diarrhoea virus epidemic in North America. Since the identification of this novel risk factor, scientists have conducted numerous studies to understand its relevance. Over the past few years, the body of scientific evidence supporting the reality of this risk has grown substantially. In addition, numerous papers describing actions and interventions designed to mitigate this risk have been published. Therefore, the purpose of this paper is to review the literature on the risk of feed (what do we know) and the protocols developed to reduce this risk (what do we do) in an effort to develop a comprehensive document to raise awareness, facilitate learning, improve the accuracy of risk assessments and to identify knowledge gaps for future studies.

Correction: Survival of viral pathogens in animal feed ingredients under transboundary shipping models.

[This corrects the article DOI: 10.1371/journal.pone.0194509.].

Correction: Survival of viral pathogens in animal feed ingredients under transboundary shipping models.

[This corrects the article DOI: 10.1371/journal.pone.0194509.].

Survival of viral pathogens in animal feed ingredients under transboundary shipping models.

The goal of this study was to evaluate survival of important viral pathogens of livestock in animal feed ingredients imported daily into the United States under simulated transboundary conditions. Eleven viruses were selected based on global significance and impact to the livestock industry, including Foot and Mouth Disease Virus (FMDV), Classical Swine Fever Virus (CSFV), African Swine Fever Virus (ASFV), Influenza A Virus of Swine (IAV-S), Pseudorabies virus (PRV), Nipah Virus (NiV), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), Swine Vesicular Disease Virus (SVDV), Vesicular Stomatitis Virus (VSV), Porcine Circovirus Type 2 (PCV2) and Vesicular Exanthema of Swine Virus (VESV). Surrogate viruses with similar genetic and physical properties were used for 6 viruses. Surrogates belonged to the same virus families as target pathogens, and included Senecavirus A (SVA) for FMDV, Bovine Viral Diarrhea Virus (BVDV) for CSFV, Bovine Herpesvirus Type 1 (BHV-1) for PRV, Canine Distemper Virus (CDV) for NiV, Porcine Sapelovirus (PSV) for SVDV and Feline Calicivirus (FCV) for VESV. For the remaining target viruses, actual pathogens were used. Virus survival was evaluated using Trans-Pacific or Trans-Atlantic transboundary models involving representative feed ingredients, transport times and environmental conditions, with samples tested by PCR, VI and/or swine bioassay. SVA (representing FMDV), FCV (representing VESV), BHV-1 (representing PRV), PRRSV, PSV (representing SVDV), ASFV and PCV2 maintained infectivity during transport, while BVDV (representing CSFV), VSV, CDV (representing NiV) and IAV-S did not. Notably, more viruses survived in conventional soybean meal, lysine hydrochloride, choline chloride, vitamin D and pork sausage casings. These results support published data on transboundary risk of PEDV in feed, demonstrate survival of certain viruses in specific feed ingredients ("high-risk combinations") under conditions simulating transport between continents and provide further evidence that contaminated feed ingredients may represent a risk for transport of pathogens at domestic and global levels.

Epidemiological study of air filtration systems for preventing PRRSV infection in large sow herds.

Porcine reproductive and respiratory syndrome virus (PRRSV) is the most economically significant pathogen in the US swine industry. Aerosol transmission among herds is a major concern in pig dense regions and filtration of incoming air, in combination with standard biosecurity procedures, has been demonstrated to prevent transmission of PRRSV into susceptible herds. To quantify the impact of air filtration on reducing risk of PRRSV outbreaks, we compared the incidence rate of new PRRSV introductions in 20 filtered and 17 non-filtered control sow herds in a swine dense region of North America during a 7 year study period. Events of novel virus introduction were ascertained by phylogenetic analysis of PRRSV ORF5 gene sequences. Putative new viruses were defined as exogenous (introduced) based on ORF5 nucleotide sequence differences compared to previous farm isolates. The influence of sequence difference cut-off values ranging from 2 to 10% on case definition and relative risk were evaluated. Non-filtered farms incurred about 0.5 outbreaks per year, with a seasonal increase in risk in cooler periods. Baseline risk, prior to filtration, in treatment farms was approximately 0.75 per year, approximately 50% higher than in control farms. Air filtration significantly reduced risk of PRRSV introduction events to 0.06-0.22 outbreaks per year, depending on the cut-off values used to classify a virus isolate as new to the herd. Overall, air filtration led to an approximately 80% reduction in risk of introduction of novel PRRSV, indicating that on large sow farms with good biosecurity in swine-dense regions, approximately four-fifths of PRRSV outbreaks may be attributable to aerosol transmission.

Financial implications of installing air filtration systems to prevent PRRSV infection in large sow herds.

Air filtration systems implemented in large sow herds have been demonstrated to decrease the probability of having a porcine reproductive and respiratory syndrome virus (PRRSV) outbreak. However, implementation of air filtration represents a considerable capital investment, and does not eliminate the risk of new virus introductions. The specific objectives of the study were: 1) to determine productivity differences between a cohort of filtered and non-filtered sow farms; and 2) to employ those productivity differences to model the profitability of filtration system investments in a hypothetical 3000 sow farm. Variables included in the study were production variables (quarterly) from respective herds; air filtration status; number of pig sites within 4.7 km of the farm; occurrence of a PRRSV outbreak in a quarter, and season. For the investment analyses, three Scenarios were compared in a deterministic spreadsheet model of weaned pig cost: (1) control, (2) filtered conventional attic, and (3) filtered tunnel ventilation. Model outputs indicated that a filtered farm produced 5927 more pigs than unfiltered farms. The payback periods for the investments, were estimated to be 5.35 years for Scenario 2 and 7.13 years for Scenario 3 based solely on sow herd productivity. Payback period sensitivity analyses were performed for both biological and financial inputs. The payback period was most influenced by the premium for weaned pig sales price for PRRSV-negative pigs, and the relative proportions of time that filtered vs. unfiltered farms produced PRRSV-negative pigs. A premium of $5 per pig for PRRS-negative weaned pigs reduced the estimated payback periods to 2.1 years for Scenario 2 and 2.8 years for Scenario 3.

Relationship between airborne detection of influenza A virus and the number of infected pigs.

Influenza A virus infects a wide range of species including both birds and mammals (including humans). One of the key routes by which the virus can infect populations of animals is by aerosol transmission. This study explored the relationship between number of infected pigs and the probability of detecting influenza virus RNA in bioaerosols through the course of an acute infection. Bioaerosols were collected using a cyclonic collector in two groups of 7 week-old pigs that were experimentally infected by exposure with a contact infected pig (seeder pig). After contact exposure, individual pig nasal swab samples were collected daily and air samples were collected three times per day for 8 days. All samples were tested for influenza by real-time reverse transcriptase (RRT)-PCR targeting the influenza virus matrix gene. All pigs' nasal swabs became influenza virus RRT-PCR positive upon exposure to the infected seeder pig. Airborne influenza was detected in 28/43 (65%) air samples. The temporal dynamics of influenza virus detection in air samples was in close agreement with the nasal shedding pattern in the infected pigs. First detection of positive bioaerosols happened at 1 day post contact (DPC). Positive bioaerosols were consistently detected between 3 and 6 DPC, a time when most pigs were also shedding virus in nasal secretions. Overall, the odds of detecting a positive air sample increased 2.2 times for every additional nasal swab positive pig in the group. In summary, there was a strong relationship between the number of pigs shedding influenza virus in nasal secretions and the generation of bioaerosols during the course of an acute infection.

Effect of modified-live porcine reproductive and respiratory syndrome virus (PRRSv) vaccine on the shedding of wild-type virus from an infected population of growing pigs.

There are ongoing efforts to eliminate porcine reproductive and respiratory syndrome virus (PRRSv) from regions in the United States swine industry. However, an important challenge for the accomplishment of those efforts is the re-infection of pig units due to the area spread of PRRSv. The objective of this study was to evaluate the effect of PRRS modified-live virus vaccine (MLV) on viral shedding and on dynamics of PRRSv infection in pig populations raised under commercial conditions. The study composed of two rooms of 1000 pigs each. Ten percent of pigs of each room were inoculated with a field isolate of PRRSv. Rooms had separate air spaces and strict scientifically validated biosecurity protocols were adopted to avoid movement of pathogens between rooms. At 8 and 36 dpi (days post inoculation), all pigs of the challenge-vaccine group were inoculated with a MLV vaccine. Pigs of the challenge-control group were placebo-inoculated. Blood and oral fluid samples were collected from each room at 0, 8, 36, 70, 96 and 118 dpi for PRRSv RNA detection using PCR. PRRSv-antibodies were also screened from blood serum samples with a commercially available ELISA test. Additionally, tonsil scraping samples were collected from both groups at 70, 96 and 118 dpi. Moreover, air samples were collected 6 times per week from 0 to 118 dpi and were tested for PRRSv RNA using qPCR assay. There was no difference in the PRRSv infection dynamics measured as duration and magnitude of viremia and seroconversion. Also, there was no difference in the frequency of tonsil scraping samples PRRSv-positive by PCR. However, the challenge-vaccine group had significantly less PRRSv shed compared to the challenge-control group. The challenge-vaccine group had significant less PRRSv-positive oral fluids at 36 dpi. Moreover, the challenge-vaccine group had significant reduction in the cumulative PRRSv shed in the air.

Infection dynamics and clinical manifestations following experimental inoculation of gilts at 90 days of gestation with a low dose of porcine reproductive and respiratory syndrome virus.

Understanding the dynamics of porcine reproductive and respiratory syndrome virus (PRRSV) vertical transmission is important to enhance the accuracy of monitoring protocols for endemically infected breeding herds. The objectives of this study were to determine the prevalence of PRRSV within infected litters, to quantify viremia, and to identify specific attributes of infected individuals. Eight gilts were intramuscularly inoculated with 10(1) TCID(50) of a mildly virulent PRRSV strain (MN-30100) at 90 d of gestation. All inoculated gilts transmitted the virus in utero. The proportion of PRRSV PCR-positive piglets and the level of viremia in the piglets were higher at 4 d of age than at birth or at weaning. No specific attributes were associated with PRRSV infection in the piglets. This is the first report, that we are aware of, documenting the efficient in utero transmission of an extremely low dose of a mildly virulent strain of PRRSV. The results support the sampling of piglets late during lactation as a tool to monitor PRRSV shedding from sow-herds.

Antigen-specific B-cell responses to porcine reproductive and respiratory syndrome virus infection.

Porcine reproductive and respiratory syndrome virus (PRRSV) causes an acute, viremic infection of 4 to 6 weeks, followed by a persistent infection lasting for several months. We characterized antibody and B-cell responses to viral proteins in acute and persistent infection to better understand the immunological basis of the prolonged infection. The humoral immune response to PRRSV was robust overall and varied among individual viral proteins, with the important exception of a delayed and relatively weak response to envelope glycoprotein 5 (GP5). Memory B cells were in secondary lymphoid organs, not in bone marrow or Peyer's patches, in contrast to the case for many mammalian species. Potent anti-PRRSV memory responses were elicited to recall antigen in vitro, even though a second infection did not increase the B-cell response in vivo, suggesting that productive reinfection does not occur in vivo. Antibody titers to several viral proteins decline over time, even though abundant antigen is known to be present in lymphoid tissues, possibly indicating ineffective antigen presentation. The appearance of antibodies to GP5 is delayed relative to the resolution of viremia, suggesting that anti-GP5 antibodies are not crucial for resolving viremia. Lastly, viral infection had no immunosuppressive effect on the humoral response to a second, unrelated antigen. Taking these data together, the active effector and memory B-cell responses to PRRSV are robust, and over time the humoral immune response to PRRSV is effective. However, the delayed response against GP5 early in infection may contribute to the prolonged acute infection and the establishment of persistence.