Investigation of the distance to slaughterhouses and weather parameters in the occurrence of porcine reproductive and respiratory syndrome outbreaks in U.S. swine breeding herds.

Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically important diseases affecting the swine industry. The main objective of this study was to assess whether sow farm distance to slaughterhouses and meteorological variables were associated with PRRS outbreaks. This case-control study paired 104 sow farms with or without a reported PRRS outbreak (N = 208) during the same period. Data on the distance to the closest slaughterhouse, swine density, presence of an air filtration system, or a neighboring farm, and weather conditions were collected, and a multivariable conditional logistic regression model was created to investigate the association between variables of interest and the occurrence of a PRRS outbreak. Swine density, presence of an air filtration system, presence of a neighboring farm, and PRRS herd-level status before the outbreak were associated with the occurrence of PRRS outbreaks. Farms in areas with higher swine density and nearby swine farms had increased odds of reporting an outbreak compared to farms in low swine density areas and farms having no neighbors. Under the conditions of this study, none of the meteorological variables or the distance to the closest slaughterhouse were associated with occurrence of PRRS outbreaks.

Enquête sur la distance aux abattoirs et les paramètres météorologiques dans l'apparition d'épidémies de syndrome reproducteur et respiratoire porcin dans les troupeaux reproducteurs de porcs aux États-Unis. Le syndrome reproducteur et respiratoire porcin (SRRP) est l'une des maladies les plus importantes sur le plan économique affectant l'industrie porcine. L'objectif principal de cette étude était d'évaluer si la distance entre les élevages de truies et les abattoirs et les variables météorologiques étaient associées aux épidémies de SRRP. Cette étude cas-témoin a apparié 104 élevages de truies avec ou sans éclosion de SRRP déclarée (N = 208) au cours de la même période. Des données sur la distance à l'abattoir le plus proche, la densité porcine, la présence d'un système de filtration d'air ou d'une ferme voisine et les conditions météorologiques ont été recueillies, et un modèle de régression logistique conditionnelle multivariable a été créé pour étudier l'association entre les variables d'intérêt et l'occurrence d'une épidémie de SRRP.La densité porcine, la présence d'un système de filtration de l'air, la présence d'une ferme voisine et le statut du troupeau relativement au SRRP avant l'épidémie ont été associés à l'apparition d'épidémies de SRRP. Les fermes situées dans des zones à forte densité porcine et d'autres fermes porcines à proximité avaient plus de chances de signaler une épidémie que les fermes situées dans des zones à faible densité porcine et les fermes sans voisins. Dans les conditions de cette étude, aucune des variables météorologiques ni la distance à l'abattoir le plus proche n'étaient associées à la survenue d'épidémies de SRRP.(Traduit par Dr Serge Messier).

Benefit-cost analysis to estimate the payback time and the economic value of two Mycoplasma hyopneumoniae elimination methods in breeding herds.

Mycoplasma hyopneumoniae (Mhyo) is generally accepted to be the most common porcine respiratory pathogen worldwide causing big economical losses in swine production by affecting pig's downstream performance. The objective of this study was to develop a partial budget model to determine the payback period and economic value of two Mhyo elimination protocols. Retrospective data recorded from 2004 to 2017 from 70 breeding herds that implemented herd closure or whole-herd medication protocol targeting Mhyo elimination. Close out data was used to estimate differences in downstream performance between Mhyo-negative and positive flows. Assuming a 5000 sows breed-to-finish operation producing 135,870 weaned pigs and 125,000 finishing pigs/year, the total cost for implementing Mhyo elimination was $112,100 using the herd closure protocol, and $185,700 for the medication protocol. Statistically differences (p < 0.05) in downstream performance were observed for ADG and mortality, but not for feed conversion rate. The parameters that accounts for the greatest benefits were related to the improvement in ADG, savings in antibiotic medication in growing pigs and improvement in feed conversion rate. The benefit of Mhyo elimination was $877,375 per farm per year, or $7.00 per pig marketed. The estimated project value after 1 year was $616,121 for the herd closure considering a probability of success of 83%, and $323,177 for the medication protocol for 58% chance of success. The project value reached the break-even point when the cost per sow was $145.64 for the herd closure and $101.78 for the medication protocol. The payback period was 2 months after the start of marketing Mhyo-negative pigs for the herd closure, and 7 months for the medication protocol adjusted for the probability of success for each protocol. The protocols described here can be easily applied with a good success rate and showing that the benefits obtained are greater than the costs of project failure. Even if the farm stayed negative only a year, the economic benefits downstream are worth the investment. This information may help producers and veterinarians on decision-making process to conduct a Mhyo elimination protocol in their herds.

Effect of litter aggregation and pooling on detection of porcine reproductive and respiratory virus in piglet processing fluids.

A sampling technique has been validated to monitor porcine reproductive and respiratory syndrome virus 2 (PRRSV-2) using the serosanguinous exudate known as processing fluids (PFs) that accumulate from tissues obtained during tail docking and castration. PFs are an aggregate sample of large numbers of piglets and litters. However, little is known about the effect of litter aggregation on the ability of PCR to correctly classify an aggregated PF sample as positive. We evaluated both the effect of litter aggregation and of PF pooling on PCR detection. We estimated that aggregation of at least 50 litters was possible when a pig with a Ct value of ~22 was present in the sample, and aggregation of up to 40 litters was possible when there was a sample with a Ct value of ~33. Pooling did not affect PCR detection when initial Ct values of 20 and 25 were assessed. However, in litters with initial Ct values of ≥30, the amount of pooling should be reduced. Our results provide producers and practitioners with a general framework to interpret more accurately the results of their PRRSV-2 surveillance programs using PF.

Production Losses From an Endemic Animal Disease: Porcine Reproductive and Respiratory Syndrome (PRRS) in Selected Midwest US Sow Farms.

Porcine reproductive and respiratory syndrome (PRRS) is an endemic disease causing important economic losses to the US swine industry. The complex epidemiology of the disease, along with the diverse clinical outputs observed in different types of infected farms, have hampered efforts to quantify PRRS' impact on production over time. We measured the impact of PRRS on the production of weaned pigs using a log-linear fixed effects model to evaluate longitudinal data collected from 16 sow farms belonging to a specific firm. We measured seven additional indicators of farm performance to gain insight into disease dynamics. We used pre-outbreak longitudinal data to establish a baseline that was then used to estimate the decrease in production. A significant rise of abortions in the week before the outbreak was reported was the strongest signal of PRRSV activity. In addition, production declined slightly one week before the outbreak and then fell markedly until weeks 5 and 6 post-outbreak. Recovery was not monotonic, cycling gently around a rising trend. At the end of the study period (35 weeks post-outbreak), neither the production of weaned pigs nor any of the performance indicators had fully recovered to baseline levels. This result suggests PRSS outbreaks may last longer than has been found in most other studies. We assessed PRRS' effect on farm efficiency as measured by changes in sow production of weaned pigs per year. We translated production losses into revenue losses assuming an average market price of $45.2/weaned pig. We estimate that the average PRSS outbreak reduced production by approximately 7.4%, relative to annual output in the absence of an outbreak. PRRS reduced production by 1.92 weaned pigs per sow when adjusted to an annual basis. This decrease is substantially larger than the 1.44 decrease of weaned pigs per sow/year reported elsewhere.

Surveillance of porcine reproductive and respiratory syndrome virus in the United States using risk mapping and species distribution modeling.

Porcine reproductive and respiratory syndrome virus (PRRSv) outbreaks cause significant financial losses to the U.S. swine industry, where the pathogen is endemic. Seasonal increases in the number of outbreaks are typically observed using PRRSv epidemic curves. However, the nature and extent to which demographic and environmental factors influence the risk for PRRSv outbreaks in the country remains unclear. The objective of this study was to develop risk maps for PRRSv outbreaks across the United States (U.S.) and compare ecological dynamics of the disease in five of the most important swine production regions of the country. This study integrates spatial information regarding PRRSv surveillance with relevant demographic and environmental factors collected between 2009 and 2016. We used presence-only Maximum Entropy (Maxent), a species distribution modeling approach, to model the spatial risk of PRRSv in swine populations. Data fitted the selected model relatively well when the modeling approach was conducted by region (training and testing AUCs<0.75). All of the Maxent models selected identified high-risk areas, with probabilities greater than 0.5. The relative contribution of pig density to PRRSv risk was highest in pig-densely populated areas (Minnesota, Iowa and North Carolina), whereas climate and land cover were important in areas with relatively low pig densities (Illinois, Indiana, South Dakota, Nebraska, Kansas, Oklahoma, Colorado, and Texas). Although many previous studies associated the risk of PRRSv with high pig density and climatic factors, the study here quantifies, for the first time in the peer-reviewed literature, the spatial variation and relative contribution of these factors across different swine production regions in the U.S. The results will help in the design and implement of early detection, prevention, and control strategies for one of the most devastating diseases affecting the swine industry in the U.S.

Influenza Herd-Level Prevalence and Seasonality in Breed-to-Wean Pig Farms in the Midwestern United States.

Influenza is a costly disease for pig producers and understanding its epidemiology is critical to control it. In this study, we aimed to estimate the herd-level prevalence and seasonality of influenza in breed-to-wean pig farms, evaluate the correlation between influenza herd-level prevalence and meteorological conditions, and characterize influenza genetic diversity over time. A cohort of 34 breed-to-wean farms with monthly influenza status obtained over a 5-year period in piglets prior to wean was selected. A farm was considered positive in a given month if at least one oral fluid tested influenza positive by reverse transcriptase polymerase chain reaction. Influenza seasonality was assessed combining autoregressive integrated moving average (ARIMA) models with trigonometric functions as covariates. Meteorological conditions were gathered from local land-based weather stations, monthly aggregated and correlated with influenza herd-level prevalence. Influenza herd-level prevalence had a median of 28% with a range from 7 to 57% and followed a cyclical pattern with levels increasing during fall, peaking in both early winter (December) and late spring (May), and decreasing in summer. Influenza herd-level prevalence was correlated with mean outdoor air absolute humidity (AH) and temperature. Influenza genetic diversity was substantial over time with influenza isolates belonging to 10 distinct clades from which H1 delta 1 and H1 gamma 1 were the most common. Twenty-one percent of farms had three different clades co-circulating over time, 18% of farms had two clades, and 41% of farms had one clade. In summary, our study showed that influenza had a cyclical pattern explained in part by air AH and temperature changes over time, and highlighted the importance of active surveillance to identify high-risk periods when strategic control measures for influenza could be implemented.

Translating Big Data into Smart Data for Veterinary Epidemiology.

The increasing availability and complexity of data has led to new opportunities and challenges in veterinary epidemiology around how to translate abundant, diverse, and rapidly growing "big" data into meaningful insights for animal health. Big data analytics are used to understand health risks and minimize the impact of adverse animal health issues through identifying high-risk populations, combining data or processes acting at multiple scales through epidemiological modeling approaches, and harnessing high velocity data to monitor animal health trends and detect emerging health threats. The advent of big data requires the incorporation of new skills into veterinary epidemiology training, including, for example, machine learning and coding, to prepare a new generation of scientists and practitioners to engage with big data. Establishing pipelines to analyze big data in near real-time is the next step for progressing from simply having "big data" to create "smart data," with the objective of improving understanding of health risks, effectiveness of management and policy decisions, and ultimately preventing or at least minimizing the impact of adverse animal health issues.

A Review of Quantitative Tools Used to Assess the Epidemiology of Porcine Reproductive and Respiratory Syndrome in U.S. Swine Farms Using Dr. Morrison's Swine Health Monitoring Program Data.

Porcine reproductive and respiratory syndrome (PRRS) causes far-reaching financial losses to infected countries and regions, including the U.S. The Dr. Morrison's Swine Health Monitoring Program (MSHMP) is a voluntary initiative in which producers and veterinarians share sow farm PRRS status weekly to contribute to the understanding, in quantitative terms, of PRRS epidemiological dynamics and, ultimately, to support its control in the U.S. Here, we offer a review of a variety of analytic tools that were applied to MSHMP data to assess disease dynamics in quantitative terms to support the decision-making process for veterinarians and producers. Use of those methods has helped the U.S. swine industry to quantify the cyclical patterns of PRRS, to describe the impact that emerging pathogens has had on that pattern, to identify the nature and extent at which environmental factors (e.g., precipitation or land cover) influence PRRS risk, to identify PRRS virus emerging strains, and to assess the influence that voluntary reporting has on disease control. Results from the numerous studies reviewed here provide important insights into PRRS epidemiology that help to create the foundations for a near real-time prediction of disease risk, and, ultimately, will contribute to support the prevention and control of, arguably, one of the most devastating diseases affecting the North American swine industry. The review also demonstrates how different approaches to analyze and visualize the data may help to add value to the routine collection of surveillance data and support infectious animal disease control.

Novel approaches for Spatial and Molecular Surveillance of Porcine Reproductive and Respiratory Syndrome Virus (PRRSv) in the United States.

The US swine industry has been impaired over the last 25 years by the far-reaching financial losses caused by the porcine reproductive and respiratory syndrome (PRRS). Here, we explored the relations between the spatial risk of PRRS outbreaks and its phylodynamic history in the U.S during 1998-2016 using ORF5 sequences collected from swine farms in the Midwest region. We used maximum entropy and Bayesian phylodynamic models to generate risk maps for PRRS outbreaks and reconstructed the evolutionary history of three selected phylogenetic clades (A, B and C). High-risk areas for PRRS were best-predicted by pig density and climate seasonality and included Minnesota, Iowa and South Dakota. Phylodynamic models demonstrated that the geographical spread of the three clades followed a heterogeneous spatial diffusion process. Furthermore, PRRS viruses were characterized by typical seasonality in their population size. However, endemic strains were characterized by a substantially slower population growth and evolutionary rates, as well as smaller spatial dispersal rates when compared to emerging strains. We demonstrated the prospects of combining inferences derived from two unique analytical methods to inform decisions related to risk-based interventions of an important pathogen affecting one of the largest food animal industries in the world.

Estimation of Time-Dependent Reproduction Numbers for Porcine Reproductive and Respiratory Syndrome across Different Regions and Production Systems of the US.

Porcine reproductive and respiratory syndrome (PRRS) is, arguably, the most impactful disease for the North American swine industry, due to its known considerable economic losses. The Swine Health Monitoring Project (SHMP) monitors and reports weekly new PRRS cases in 766 sow herds across the US. The time-dependent reproduction number (TD-R) is a measure of a pathogen's transmissibility. It may serve to capture and report PRRS virus (PRRSV) spread at the regional and system levels. The primary objective of the study here was to estimate the TD-R values for PRRSV using regional and system-level PRRS data, and to contrast it with commonly used metrics of disease, such as incidence estimates and space-time clusters. The second objective was to test whether the estimated TD-Rs were homogenous across four US regions. Retrospective monthly incidence data (2009-2016) were available from the SHMP. The dataset was divided into four regions based on location of participants, and demographic and environmental features, namely, South East (North Carolina), Upper Midwest East (UME, Minnesota/Iowa), Upper Midwest West (Nebraska/South Dakota), and South (Oklahoma panhandle). Generation time distributions were fit to incidence data for each region, and used to calculate the TD-Rs. The Kruskal-Wallis test was used to determine whether the median TD-Rs differed across the four areas. Furthermore, we used a space-time permutation model to assess spatial-temporal patterns for the four regions. Results showed TD-Rs were right skewed with median values close to "1" across all regions, confirming that PRRS has an overall endemic nature. Variation in the TD-R patterns was noted across regions and production systems. Statistically significant periods of PRRSV spread (TD-R > 1) were identified for all regions except UME. A minimum of three space-time clusters were detected for all regions considering the time period examined herein; and their overlap with "spreader events" identified by the TD-R method varied according to region. TD-Rs may help to measure PRRS spread to understand, in quantitative terms, disease spread, and, ultimately, support the design, implementation, and monitoring of interventions aimed at mitigating the impact of PRRSV spread in the US.

Evaluation of an electrostatic particle ionization technology for decreasing airborne pathogens in pigs.

Influenza A virus (IAV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine epidemic diarrhea virus (PEDV) and Staphylococcus aureus are important swine pathogens capable of being transmitted via aerosols. The electrostatic particle ionization system (EPI) consists of a conductive line that emits negative ions that charge particles electrically resulting in the settling of airborne particles onto surfaces and potentially decreasing the risk of pathogen dissemination. The objectives of this study were to determine the effect of the EPI system on the quantity and viability of IAV, PRRSV, PEDV and S. aureus in experimentally generated aerosols and in aerosols generated by infected animals. Efficiency at removing airborne particles was evaluated as a function of particle size (ranging from 0.4 to 10 µm), distance from the source of ions (1, 2 and 3 m) and relative air humidity (RH 30 vs. 70 %). Aerosols were sampled with the EPI system "off" and "on." Removal efficiency was significantly greater for all pathogens when the EPI line was the closest to the source of aerosols. There was a greater reduction for larger particles ranging between 3.3 and 9 µm, which varied by pathogen. Overall airborne pathogen reduction ranged between 0.5 and 1.9 logs. Viable pathogens were detected with the EPI system "on," but there was a trend to reducing the quantity of viable PRRSV and IAV. There was not a significant effect on the pathogens removal efficiency based on the RH conditions tested. In summary, distance to the source of ions, type of pathogen and particle size influenced the removal efficiency of the EPI system. The reduction in infectious agents in the air by the EPI technology could potentially decrease the microbial exposure for pigs and people in confinement livestock facilities.

Applications of Bayesian Phylodynamic Methods in a Recent U.S. Porcine Reproductive and Respiratory Syndrome Virus Outbreak.

Classical phylogenetic methods such as neighbor-joining or maximum likelihood trees, provide limited inferences about the evolution of important pathogens and ignore important evolutionary parameters and uncertainties, which in turn limits decision making related to surveillance, control, and prevention resources. Bayesian phylodynamic models have recently been used to test research hypotheses related to evolution of infectious agents. However, few studies have attempted to model the evolutionary dynamics of porcine reproductive and respiratory syndrome virus (PRRSV) and, to the authors' knowledge, no attempt has been made to use large volumes of routinely collected data, sometimes referred to as big data, in the context of animal disease surveillance. The objective of this study was to explore and discuss the applications of Bayesian phylodynamic methods for modeling the evolution and spread of a notable 1-7-4 RFLP-type PRRSV between 2014 and 2015. A convenience sample of 288 ORF5 sequences was collected from 5 swine production systems in the United States between September 2003 and March 2015. Using coalescence and discrete trait phylodynamic models, we were able to infer population growth and demographic history of the virus, identified the most likely ancestral system (root state posterior probability = 0.95) and revealed significant dispersal routes (Bayes factor > 6) of viral exchange among systems. Results indicate that currently circulating viruses are evolving rapidly, and show a higher level of relative genetic diversity over time, when compared to earlier relatives. Biological soundness of model results is supported by the finding that sow farms were responsible for PRRSV spread within the systems. Such results cannot be obtained by traditional phylogenetic methods, and therefore, our results provide a methodological framework for molecular epidemiological modeling of new PRRSV outbreaks and demonstrate the prospects of phylodynamic models to inform decision-making processes for routine surveillance and, ultimately, to support prevention and control of food animal disease at local and regional scales.

Production impact & time to stability in sow herds infected with porcine epidemic diarrhea virus (PEDV).

PEDV was first detected in United States in May, 2013. The virus spread through the swine industry and was reported in 30 US states by June, 2014 (Morrison and Goede, 2014). There are limited data describing the impact on production in sow farms. Veterinarians attempt to control the virus in sow herds with a program that stimulates herd immunity. There are no data on how long it takes with this control program to achieve a stable state of consistently produce weaned pigs that are not infected with the virus. This study involved participants and data from an existing program called the Swine Health Monitoring Project. Veterinarians were invited to share production data from 429 herds infected with PEDV. These data, in conjunction with diagnostic reports, were used to estimate the time required for the herd to produce PEDV PCR negative pigs and the production loss. Of the 429 infected herds that achieved the stable state of weaning PEDV PCR negative pigs, the median time was 28 weeks, ranging from 7 to 64 weeks. A median of 2.7 piglets/inventoried sow were not weaned and the average time required to recover to baseline production was 10 weeks in 183 herds. Herd infected in quarters 3 or 4 of the year had approximately twice the negative impact. These data are valuable for veterinarians in advising clients on the anticipated impact and time to re-achieve a stable state with regards to PEDV.

Economic Analysis of Immunization Strategies for PRRS Control [corrected].

Porcine reproductive and respiratory syndrome virus (PRRSv) is a swine-specific pathogen that causes significant increases in production costs. When a breeding herd becomes infected, in an attempt to hasten control and elimination of PRRSv, some veterinarians have adopted a strategy called load-close-expose which consists of interrupting replacement pig introductions into the herd for several weeks (herd closure) and exposing the whole herd to a replicating PRRSv to boost herd immunity. Either modified-live virus (MLV) vaccine or live field-virus inoculation (FVI) is used. This study consisted of partial budget analyses to compare MLV to FVI as the exposure method of load-close-expose program to control and eliminate PRRSv from infected breeding herds, and secondly to estimate benefit / cost of vaccinating sow herds preventatively. Under the assumptions used in this study, MLV held economic advantage over FVI. However, sensitivity analysis revealed that decreasing margin over variable costs below $ 47.32, or increasing PRRSv-attributed cost above $18.89 or achieving time-to-stability before 25 weeks resulted in advantage of FVI over MLV. Preventive vaccination of sow herds was beneficial when the frequency of PRRSv infection was at least every 1 year and 9 months [corrected]. The economics of preventative vaccination was minimally affected by cost attributed to field-type PRRSv infection on growing pigs or by the breeding herd productivity level. The models developed and described in this paper provide valuable tools to assist veterinarians in their efforts to control PRRSv.

Temporal and spatial dynamics of porcine reproductive and respiratory syndrome virus infection in the United States.

OBJECTIVE: To measure incidence and estimate temporal and spatial dynamics of porcine reproductive and respiratory syndrome virus (PRRSV) infection in US sow herds. ANIMALS: 371 sow herds in the United States from 14 production companies. PROCEDURES: The exponentially weighted moving average was used to monitor incident PRRSV infections for onset of an epidemic. The spatial scan statistic was used to identify areas at significantly high risk of PRRS epidemics. A χ(2) test was used to estimate whether there were significant differences in the quarterly and annual PRRS incidence among time periods, and a bivariable logistic regression model was used to estimate whether PRRSV infection during a given year increased the odds of that herd being infected in the following year. RESULTS: During the 4-year period of this study, 29% (91/319; 2009 to 2010), 33% (106/325; 2010 to 2011), 38% (135/355; 2011 to 2012), and 32% (117/371; 2012 to 2013) of the herds reported new infections. Weekly incidence was low during spring and summer and high during fall and winter. The exponentially weighted moving average signaled the onset of a PRRSV epidemic during the middle 2 weeks of October each year. Disease incidence was spatially clustered. Infection in the previous year increased the odds of infection in 2010 to 2011 and 2011 to 2012. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated a striking repeatability in annual PRRSV temporal and spatial patterns across 4 years of data among herds from 14 production companies, which suggested that efforts to control PRRSV at a regional level should continue to be supported.

Evidence of infectivity of airborne porcine epidemic diarrhea virus and detection of airborne viral RNA at long distances from infected herds.

Porcine epidemic diarrhea virus (PEDV) spread rapidly after being diagnosed in the USA in April 2013. In this study we assessed whether PEDV could become airborne and if so, whether the virus was infectious. Air samples were collected both from a room containing experimentally infected pigs and at various distances from the outside of swine farms experiencing acute PEDV outbreaks. Results indicated presence of infectious PEDV in the air from experimentally infected pigs and genetic material of PEDV was detected up to 10 miles downwind from naturally infected farms. Airborne transmission should be considered as a potential route for PEDV dissemination.

Airborne detection and quantification of swine influenza a virus in air samples collected inside, outside and downwind from swine barns.

Airborne transmission of influenza A virus (IAV) in swine is speculated to be an important route of virus dissemination, but data are scarce. This study attempted to detect and quantify airborne IAV by virus isolation and RRT-PCR in air samples collected under field conditions. This was accomplished by collecting air samples from four acutely infected pig farms and locating air samplers inside the barns, at the external exhaust fans and downwind from the farms at distances up to 2.1 km. IAV was detected in air samples collected in 3 out of 4 farms included in the study. Isolation of IAV was possible from air samples collected inside the barn at two of the farms and in one farm from the exhausted air. Between 13% and 100% of samples collected inside the barns tested RRT-PCR positive with an average viral load of 3.20E+05 IAV RNA copies/m³ of air. Percentage of exhaust positive air samples also ranged between 13% and 100% with an average viral load of 1.79E+04 RNA copies/m³ of air. Influenza virus RNA was detected in air samples collected between 1.5 and 2.1 Km away from the farms with viral levels significantly lower at 4.65E+03 RNA copies/m³. H1N1, H1N2 and H3N2 subtypes were detected in the air samples and the hemagglutinin gene sequences identified in the swine samples matched those in aerosols providing evidence that the viruses detected in the aerosols originated from the pigs in the farms under study. Overall our results indicate that pigs can be a source of IAV infectious aerosols and that these aerosols can be exhausted from pig barns and be transported downwind. The results from this study provide evidence of the risk of aerosol transmission in pigs under field conditions.

Control of the PI3K/Akt pathway by porcine reproductive and respiratory syndrome virus.

Phosphatidylinositol-3-kinase (PI3K)/Akt is an important cellular pathway that has been shown to participate in various replication steps of multiple viruses. In the present study, we compared the phosphorylation status of Akt during infection of MARC-145 cells and porcine alveolar macrophages (PAMs) with highly pathogenic PRRSV (HP-PRRSV) strain HuN4. We observed that biphasic activation of Akt was induced in at both the early stage (5, 15 and 30 min postinfection) and the late stage (12 and 24 h postinfection) of HP-PRRSV infection of MARC-145 cells, while an early-phase activation of Akt was found exclusively in virus-infected PAMs in vitro. Analysis with the PI3K-specific inhibitor LY294002 confirmed that PI3K acted as the upstream activator for the virus-induced activation of Akt. UV-irradiation-inactivated virus still induced the early event in PAMs but not in MARC-145 cells, suggesting that different mechanisms are employed for the early-stage induction of phosphorylated Akt within different cell cultures. We further demonstrated that FoxO1 and Bad, which serve as downstream targets of Akt, were phosphorylated in virus-infected MARC-145 cells. Moreover, the suppression of phosphorylated Akt with LY294002 significantly inhibited the virus-induced cytopathic effect (CPE) on MARC-145 cells, but it had a negligible effect on virus propagation. Collectively, our data provide new evidence of a novel role for the PI3K/Akt pathway in PRRSV infection of MARC-145 cells.