Improvements in swine herd biosecurity reduce the incidence risk of porcine reproductive and respiratory syndrome virus in breeding herds in the Midwestern United States.

OBJECTIVE: Porcine reproductive and respiratory syndrome (PRRS) is a significant disease of swine. The purpose of this study was to determine whether application of a comprehensive, science-based approach to breeding herd biosecurity, known as next-generation biosecurity (NGB), could reduce PRRS incidence risk across a large commercial production company. ANIMALS: Pigs (381,404 sows across 76 breeding herds). METHODS: From 2009 to 2020, the annual incidence risk of PRRS in sow farms managed by the same company averaged 33%, ranging from 20% to 50%. To measure the effect of NGB on PRRS incidence risk, a retrospective cohort study was conducted from July 1, 2021, to June 30, 2023, across breeding herds managed by the same company. During the analysis, 2 groups of herds emerged: those that implemented protocols for all phases of NGB (NGB COMPLETE), and those that implemented all described protocols of biosecurity except for air filtration (NGB INCOMPLETE). RESULTS: During the 2-year assessment period, 56 breeding herds were classified as NGB COMPLETE, while 20 herds were NGB INCOMPLETE. The PRRS incidence risk in NGB COMPLETE herds was 8.9% as compared to 40.0% in NGB INCOMPLETE herds. From disease year 1 (July 1, 2021, to June 30, 2022) and disease year 2 (July 1, 2022, to June 30, 2023), system-wide PRRS incidence risk was 8.6% and 9.2%, respectively. The association between NGB status and PRRS incidence risk for the 2-year period was statistically significant at a P value of .006. CLINICAL RELEVANCE: Results of the present report provided evidence that improvements in biosecurity result in lower PRRS incidence risk under large-scale commercial swine production conditions.

Will swine veterinarians lead by meeting the next-generation needs of our industry?

The US swine industry is currently challenged by the potential of transboundary animal disease (eg, African swine fever) entry to the national herd and the relentless pressures of domestic diseases (eg, porcine reproductive and respiratory syndrome). The task of the swine veterinarian is to biosecure both the national herd and their customers' local farms to mitigate these risks. This Viewpoint raises 4 questions that swine veterinarians, including practicing (private and corporate), industry, research, academic, and regulatory (state and federal) veterinarians who spend a portion of their time controlling, treating, preventing, or eradicating diseases of swine, must answer to meet the needs of their farms to compete globally and survive. In addition, it appears that there is sufficient science-based information to move forward in a collaborative manner and that the goals of prevention of African swine fever and elimination of porcine reproductive and respiratory syndrome virus are technically possible. Therefore, as previous generations of swine veterinarians led the US industry in the elimination of foot-and-mouth disease virus, classical swine fever virus, and pseudorabies virus from the national herd, the central challenge is whether the next generation of veterinarians will provide the necessary leadership to deal with the current industry and its next-generation challenges.

An Assessment of Diagnostic Assays and Sample Types in the Detection of an Attenuated Genotype 5 African Swine Fever Virus in European Pigs over a 3-Month Period.

African swine fever virus causes hemorrhagic disease in swine. Attenuated strains are reported in Africa, Europe, and Asia. Few studies on the diagnostic detection of attenuated ASF viruses are available. Two groups of pigs were inoculated with an attenuated ASFV. Group 2 was also vaccinated with an attenuated porcine reproductive and respiratory syndrome virus vaccine. Commercially available ELISA, as well as extraction and qPCR assays, were used to detect antibodies in serum and oral fluids (OF) and nucleic acid in buccal swabs, tonsillar scrapings, OF, and blood samples collected over 93 days, respectively. After 12 dpi, serum (88.9% to 90.9%) in Group 1 was significantly better for antibody detection than OF (0.7% to 68.4%). Group 1's overall qPCR detection was highest in blood (48.7%) and OF (44.2%), with the highest detection in blood (85.2%) from 8 to 21 days post inoculation (dpi) and in OF (83.3%) from 1 to 7 dpi. Group 2's results were not significantly different from Group 1, but detection rates were lower overall. Early detection of attenuated ASFV variants requires active surveillance in apparently healthy animals and is only reliable at the herd level. Likewise, antibody testing will be needed to prove freedom from disease.

Evaluating the effect of temperature on viral survival in plant-based feed during storage.

Viruses of veterinary significance are known to survive for extended periods in plant-based feed ingredients imported into North America. To reduce the likelihood of virus introduction, high-risk ingredients, such as oil seed meals, are stored in designated facilities for extended periods under controlled environmental conditions to minimize viral infectivity prior to use in diets. While 30 days has become a standard storage period, the required ambient temperature to inactivate viruses during this time is not known. To address the question, 1-metric tonne totes of conventional soybean meal were inoculated with PRRSV 144 lineage 1C variant and SVA prior to storage for 30 days at 23.9°C, 15.5°C or 10°C, and feeding to pigs. Virus infectivity was evaluated through detection of viral RNA in oral fluid samples, along with clinical signs. Results indicated that inactivation of both viruses occurred in soy stored at 23.9°C. In contrast, SVA infectivity was observed in soy stored at both 15.5°C and 10°C, while PRRSV 144 L1C variant infectivity was only observed in soy stored at 10°C. These results suggest that a storage period of 30 days and a temperature of 23.9°C may assist in the reduction of the risk of virus contaminated plant-based feed ingredients, such as soybean meal.

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.

Interventions to reduce porcine epidemic diarrhea virus prevalence in feed in a Chinese swine production system: A case study.

Research has shown that feed and feed ingredients can be one of the potential routes of transmitting viral pathogens into swine farms. In this short communication, we report two cases of Porcine Epidemic Diarrhea (PED) in two sow farms located in eastern China. Immediately after the outbreaks, extensive sampling and testing for genetic materials of PEDV was carried out on farms, and at the feed mill, in an effort to identify possible sources of infection based on field observations of local area viral spread and interventions already implemented to lower risk of this spread. Samples collected from personnel or supplies entering the farms were inspected and proved as low risk factors. In contrast, feed and feed ingredient samples collected at the on-farm feed bins, and at the feed mill, tested positive for PEDV RNA. Based on these data, multiple interventions to lower viral spread via feed were implemented including (1) simplification of diet formulation excluding high-risk ingredients, (2) extension of thermal treatment during pellet conditioning and (3) maximising feed quarantine on farm up to 7 days from feed delivery to consumption. Collectively, these interventions appeared to have a positive effect as the prevalence of PED-related disease and the number of PEDV-positive feed or feed ingredient samples decreased considerably following implementation.

Evidence of viral survival in representative volumes of feed and feed ingredients during long-distance commercial transport across the continental United States.

The hypothesis that feed ingredients could serve as vehicles for the transport and transmission of viral pathogens was first validated under laboratory conditions. To bridge the gap from the laboratory to the field, this current project tested whether three significant viruses of swine could survive in feed ingredients during long-distance commercial transport across the continental US. One-metric tonne totes of soybean meal (organic and conventional) and complete feed were spiked with a 10 ml mixture of PRRSV 174, PEDV and SVA and transported for 23 days in a commercial semi-trailer truck, crossing 29 states, and 10,183 km. Samples were tested for the presence of viral RNA by PCR, and for viable virus in soy-based samples by swine bioassay and in complete feed samples by natural feeding. Viable PRRSV, PEDV and SVA were detected in both soy products and viable PEDV and SVA in complete feed. These results provide the first evidence that viral pathogens of pigs can survive in representative volumes of feed and feed ingredients during long-distance commercial transport across the continental United States.

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.

Use of a demonstration project to evaluate viral survival in feed: Proof of concept.

In 2014, the hypothesis that feed ingredients could serve as vehicles for the transport and transmission of viral pathogens was proposed and evaluated by multiple investigators under laboratory conditions. In an attempt to validate these data, we used a demonstration project to test whether three significant viruses of swine could survive in feed ingredients under real-world shipping conditions. Samples of soya bean meal (organic and conventional), lysine, choline and vitamin A were spiked with a mixture of PRRSV 174, PEDV and SVA and transported for 21 days in the trailer of a commercial transport vehicle, encompassing 14 states and 9,741 km. Samples were tested for viral genome and viability at the end of the transit period. Regarding viability, PRRSV, PEDV and SVA were all detected as infectious in bioassays following inoculation with both soy products. In addition, viable PRRSV and SVA were detected by bioassay pigs inoculated with samples of vitamin A, and infectious SVA was detected in pigs inoculated with samples of lysine and choline. These results provide further evidence that select viral pathogens of pigs can survive in certain feed ingredients during commercial transit.

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.

Modeling the transboundary risk of feed ingredients contaminated with porcine epidemic diarrhea virus.

BACKGROUND: This study describes a model developed to evaluate the transboundary risk of PEDV-contaminated swine feed ingredients and the effect of two mitigation strategies during a simulated transport event from China to the US. RESULTS: Ingredients imported to the USA from China, including organic & conventional soybeans and meal, lysine hydrochloride, D-L methionine, tryptophan, Vitamins A, D & E, choline, carriers (rice hulls, corn cobs) and feed grade tetracycline, were inoculated with PEDV. Control ingredients, and treatments (ingredients plus a liquid antimicrobial (SalCURB, Kemin Industries (LA) or a 2% custom medium chain fatty acid blend (MCFA)) were tested. The model ran for 37 days, simulating transport of cargo from Beijing, China to Des Moines, IA, US from December 23, 2012 to January 28, 2013. To mimic conditions on land and sea, historical temperature and percent relative humidity (% RH) data were programmed into an environmental chamber which stored all containers. To evaluate PEDV viability over time, ingredients were organized into 1 of 4 batches of samples, each batch representing a specific segment of transport. Batch 1 (segment 1) simulated transport of contaminated ingredients from manufacturing plants in Beijing (day 1 post-contamination (PC)). Batch 2 (segments 1 and 2) simulated manufacturing and delivery to Shanghai, including time in Anquing terminal awaiting shipment (days 1-8 PC). Batch 3 (segments 1, 2 and 3) represented time in China, the crossing of the Pacific and entry to the US at the San Francisco, CA terminal (day 1-27 PC). Batch 4 (segments 1-4) represented the previous events, including transport to Des Moines, IA (days 1-37 PC). Across control (non-treated) ingredients, viable PEDV was detected in soybean meal (organic and conventional), Vitamin D, lysine hydrochloride and choline chloride. In contrast, viable PEDV was not detected in any samples treated with LA or MCFA. CONCLUSIONS: These results demonstrate the ability of PEDV to survive in a subset of feed ingredients using a model simulating shipment from China to the US. This is proof of concept suggesting that contaminated feed ingredients could serve as transboundary risk factors for PEDV, along with the identification of effective mitigation options.

Evaluation of the long-term effect of air filtration on the occurrence of new PRRSV infections in large breeding herds in swine-dense regions.

Airborne transmission of porcine reproductive and respiratory syndrome virus (PRRSV) is a risk factor for the infection of susceptible populations. Therefore, a long­term sustainability study of air filtration as a means to reduce this risk was conducted. Participating herds (n = 38) were organized into 4 independent cohorts and the effect of air filtration on the occurrence of new PRRSV infections was analyzed at 3 different levels from September 2008 to January 2012 including the likelihood of infection in contemporary filtered and non-filtered herds, the likelihood of infection before and after implementation of filtration and the time to failure in filtered and non-filtered herds. Results indicated that new PRRSV infections in filtered breeding herds were significantly lower than in contemporary non-filtered control herds (P < 0.01), the odds for a new PRRSV infection in breeding herds before filtration was 7.97 times higher than the odds after filtration was initiated (P < 0.01) and the median time to new PRRSV infections in filtered breeding herds of 30 months was significantly longer than the 11 months observed in non-filtered herds (P < 0.01). In conclusion, across all 3 levels of analysis, the long-term effect of air filtration on reducing the occurrence of new PRRSV infections in the study population was demonstrated.

Comparative infection efficiency of Porcine reproductive and respiratory syndrome virus field isolates on MA104 cells and porcine alveolar macrophages.

Isolation of Porcine reproductive and respiratory syndrome virus (PRRSV) on MA104 or MARC-145 cells is frequently used in PRRS diagnosis. However, the ability of recent field isolates to grow on these established simian cell lines has not been determined. The aim of this study was to characterize the growth of PRRSV field isolates on primary porcine alveolar macrophages (PAMs) and MA104 cells in comparison with the growth of the laboratory-adapted strain VR-2332. A cytopathic effect was observed in 70% of serum samples after 1 passage on PAMs and was verified by immunofluorescent staining or reverse transcriptase-polymerase chain reaction. Field isolate growth was observed on MA104 cells for only 1 of 50 serum samples after 14 d. Strain VR-2332 grew readily in MA104 cells [maximum titer, 10(7) TCID(50) (median tissue culture infective dose) per milliliter at 30 h] but not in PAMs (10(2) TCID(50)/mL at 72 h). These results show that PAMs are superior to simian cells for diagnostic isolation of current field PRRSV strains.