Porcine Reproductive and Respiratory Syndrome (PRRSV2) Viral Diversity within a Farrow-to-Wean Farm Cohort Study.

Describing PRRSV whole-genome viral diversity data over time within the host and within-farm is crucial for a better understanding of viral evolution and its implications. A cohort study was conducted at one naïve farrow-to-wean farm reporting a PRRSV outbreak. All piglets 3-5 days of age (DOA) born to mass-exposed sows through live virus inoculation with the recently introduced wild-type virus two weeks prior were sampled and followed up at 17-19 DOA. Samples from 127 piglets were individually tested for PRRSV by RT-PCR and 100 sequences were generated using Oxford Nanopore Technologies chemistry. Female piglets had significantly higher median Ct values than males (15.5 vs. 13.7, Kruskal-Wallis p < 0.001) at 3-5 DOA. A 52.8% mortality between sampling points was found, and the odds of dying by 17-19 DOA decreased with every one unit increase in Ct values at 3-5 DOA (OR = 0.76, 95% CI 0.61-0.94, p = 0.01). Although the within-pig percent nucleotide identity was overall high (99.7%) between 3-5 DOA and 17-19 DOA samples, ORFs 4 and 5a showed much lower identities (97.26% and 98.53%, respectively). When looking solely at ORF5, 62% of the sequences were identical to the 3-5 DOA consensus. Ten and eight regions showed increased nucleotide and amino acid genetic diversity, respectively, all found throughout ORFs 2a/2b, 4, 5a/5, 6, and 7.

Disentangling transport movement patterns of trucks either transporting pigs or while empty within a swine production system before and during the COVID-19 epidemic.

Transport of pigs between sites occurs frequently as part of genetic improvement and age segregation. However, a lack of transport biosecurity could have catastrophic implications if not managed properly as disease spread would be imminent. However, there is a lack of a comprehensive study of vehicle movement trends within swine systems in the Midwest. In this study, we aimed to describe and characterize vehicle movement patterns within one large Midwest swine system representative of modern pig production to understand movement trends and proxies for biosecurity compliance and identify potential risky behaviors that may result in a higher risk for infectious disease spread. Geolocation tracking devices recorded vehicle movements of a subset of trucks and trailers from a production system every 5 min and every time tracks entered a landmark between January 2019 and December 2020, before and during the COVID-19 pandemic. We described 6,213 transport records from 12 vehicles controlled by the company. In total, 114 predefined landmarks were included during the study period, representing 5 categories of farms and truck wash facilities. The results showed that trucks completed the majority (76.4%, 2,111/2,762) of the recorded movements. The seasonal distribution of incoming movements was similar across years (P > 0.05), while the 2019 winter and summer seasons showed higher incoming movements to sow farms than any other season, year, or production type (P < 0.05). More than half of the in-movements recorded occurred within the triad of sow farms, wean-to-market stage, and truck wash facilities. Overall, time spent at each landmark was 9.08% higher in 2020 than in 2019, without seasonal highlights, but with a notably higher time spent at truck wash facilities than any other type of landmark. Network analyses showed high connectivity among farms with identifiable clusters in the network. Furthermore, we observed a decrease in connectivity in 2020 compared with 2019, as indicated by the majority of network parameter values. Further network analysis will be needed to understand its impact on disease spread and control. However, the description and quantification of movement trends reported in this study provide findings that might be the basis for targeting infectious disease surveillance and control.

Assessing the role of sow parity on PRRSv detection by RT-qPCR through weekly processing fluids monitoring in breeding herds.

The use of processing fluids to monitor the breeding herd's porcine reproductive and respiratory syndrome (PRRS) status has gained industry acceptance. However, little is known about PRRS virus RT-qPCR detection dynamics in processing fluids and factors that may contribute to maintain PRRS virus in the herd after an outbreak. This study aimed to describe weekly RT-qPCR processing fluid results in breeding herds after an outbreak and to evaluate the proportion of RT-qPCR positive results among parity groups. Processing tissues of 15 first parity (P1), 15 second parity (P2), and 15 third parity or higher (P3+) litters (parity groups) were collected weekly for between 19 and 46 weeks in nine breeding herds. Processing fluids were aggregated, and RT-qPCR tested by parity group weekly. Additionally, a subset of 743 processing fluid samples of litters that formed 50 parity groups, as previously described, were RT-qPCR tested individually at the litter level. The agreement between RT-qPCR results of processing fluid samples of parity groups (15 litters) and results based on individual litter testing was assessed using overall percent of agreement, Kappa statistic, and McNemar test. The association between RT-qPCR results and the parity group was evaluated using a generalized estimating equations model, after accounting for the effects of sampling week, breeding herd PRRS control strategy (i.e., open to replacements v/s closed) and herd. An autoregressive correlation structure was used to account for the repeated samplings within a herd in time. The overall agreement was 98 %, and Kappa statistic 0.955 (McNemar p = 1.0). Sensitivity of parity group processing fluid samples was estimated at 100 % (95 % CI 89-100 %), while specificity was estimated at 94 % (95 % CI 71-100 %). Although P1 aggregated litters had on average a higher proportion of RT-qPCR positive results from outbreak week 25 onwards, the proportion was not significantly different to the one observed for P2 and P3+ aggregated litters (p > 0.13). Additionally, herds that interrupted gilt entry had lower odds of PRRS RT-qPCR positivity than herds that continued entering gilts (OR = 0.35, 95 % CI 0.16-0.78). PRRS virus persistence in processing fluids was not affected by the sow parity effect in most of the breeding herds studied. No evidence of disagreement between RT-qPCR results of an aggregated sample of 15 litters and those of individual litters was observed. This level of litter aggregation testing strategy may be of particular use at the last stages of an elimination program under low PRRS virus prevalence.

Porcine reproductive and respiratory syndrome prevalence and processing fluids use for diagnosis in United States breeding herds.

Introduction: Processing fluids have been recently adopted by the U.S. swine industry as a breeding herd PRRS monitoring tool due to their increased representativeness of animals within the herd. Here, we use the Morrison Swine Health Monitoring Project (MSHMP) database, representative of ~50% of the U.S. swine breeding herd, to describe processing fluids submissions for PRRS diagnosis and their relation to PRRS prevalence and time to stability over time between 2009 and 2020. Methods: An ecological time series Poisson regression modeling the number of status 1 farms and weekly percentage of processing fluids submissions for PRRS diagnosis was done. Time to stability was calculated for sites that detected a PRRS outbreak within the study period and modeled through a proportional hazards mixed effect survival model using production system as a random-effect factor and epiweek as a panel variable. Results: Processing fluids diagnosis submissions increased starting in 2017. The difference between each year's highest and lowest weekly prevalence averaged 10.9% between 2009 and 2017, whereas it averaged 5.0% in 2018-2020 period. Each year's lowest weekly prevalence ranged from 11.3 to 19.5% in 2009-2017 and from 22.4 to 29.2% in 2018-2020. We also detected an increasing proportion of breeding sites that did not reach stability within 1 year of reporting an outbreak (chi-square for trend p < 0.0001). The total time to stability was not associated with the region of the country in which the site was located, the site's air filtration status, its PRRS status before the outbreak, or the different statuses a site achieved to be classified as stable, when accounting for the production system in the multivariate model. However, a higher proportion of system-wide processing fluids use was associated with increased time to stability. Discussion: Altogether, the temporal concurrence of processing fluids used for PRRS virus monitoring suggests that the adoption of this sampling strategy may help explain the changes observed in PRRS status 1 prevalence since 2018, although further studies are still needed.

Senecavirus A seroprevalence and risk factors in United States pig farms.

Senecavirus A (SVA) is a non-enveloped, single-stranded, positive-sense RNA virus belonging to the Picornaviridae family. Senecavirus A is constantly associated with outbreaks of vesicular disease in pigs and has been reported in several countries since its first large-scale outbreak in 2014. Senecavirus A's clinical disease and lesions are indistinguishable from other vesicular foreign animal diseases (FAD). Therefore, an FAD investigation needs to be conducted for every SVA case. For this reason, SVA has been attributed as the cause of an alarming increase in the number of yearly FAD investigations performed by the United States Department of Agriculture (USDA). The objectives of this study were to estimate the seroprevalence of SVA antibodies in breeding and growing pig farms in the United States and to determine the farm-level risk factors associated with seropositivity. A total of 5,794 blood samples were collected from 98 and 95 breeding and growing pig farms in 17 states. A farm characteristics questionnaire was sent to all farms, to which 80% responded. The responses were used to conduct logistic regression analyses to assess the risk factors associated with SVA seropositivity. The estimated farm-level seroprevalences were 17.3% and 7.4% in breeding and growing pig farms, respectively. Breeding farms had 2.64 times higher odds of SVA seropositivity than growing pig farms. One key risk factor identified in breeding farms was the practice of rendering dead animal carcasses. However, the adoption of a higher number of farm biosecurity measures was associated with a protective effect against SVA seropositivity in breeding farms.

Evaluation of dam parity and internal biosecurity practices in influenza infections in piglets prior to weaning.

Influenza is an important respiratory disease of pigs and humans. Controlling influenza in pigs is challenging due to the substantial genetic diversity of influenza A virus (IAV). In this study, we assessed the impact of internal biosecurity practices directed at limiting exposure of piglets to IAV before weaning; evaluated the association of sow parity with IAV prevalence in piglets and the levels of maternally derived antibodies (MDA), and documented the frequency of detection of IAV on farmworkers' hands and the instruments used when handling pigs. The control group included litters in rooms where no specific changes were made to standard farm procedures. The treatment group included litters in rooms where no cross-fostering or nurse sows use was allowed, and where farmworkers were required to change gloves between litters when handling pigs. Both, younger (≤ Parity 3) and older parity sows (>Parity 3) were represented in all rooms included in the study. Overall, litters in the treatment group had lower IAV prevalence (29.9 %) than litters in the control group (44.2 %) (p < 0.001), and at day 8 of age the litters from the control group had 7.5 times higher IAV prevalence than the litters from the treatment group. However, at weaning differences were not found (77.2 % vs. 81 % for treatment vs. control, respectively, p = 0.41). There were no differences in IAV detection between parity groups at any of the sampling points (p = 0.86) and incidence of detection in sows from farrowing to weaning was 29 %. Piglets that tested ELISA negative were 1.3 times more likely to test IAV positive than piglets that were ELISA positive for IAV antibody test, suggesting that effective colostrum intake may reduce the likelihood of infection. IAV was detected on 46 % of the instruments used when handling piglets and on 58 % of farmworkers' hands, indicating the potential risk for mechanical transmission of IAV in pigs. Overall, we showed that the implementation of internal biosecurity practices that limit IAV exposure to newborn piglets helped delay IAV infections but were not sufficient to reduce the prevalence of IAV infection in litters at weaning.

Growing pig incidence rate, control and prevention of porcine epidemic diarrhea virus in a large pig production system in the United States.

BACKGROUND: In 2013, PEDV was introduced in the United States (U.S.) and rapidly spread across the country. Here we describe the occurrence of PEDV in the growing pig herd of one large U.S. production system through an active surveillance set in place between October 2019 and November 2020 designed to assess disease status upon placement into the growing pig site, before shipping to the slaughter plant and when diarrhea events were present at the site. We also assessed the impact of preventive procedures implemented in PEDV incidence that comprised site-specific equipment segregation and biosecurity changes regarding personnel movement between sites. RESULTS: 36.50% (100/274) of the sites had at least one PEDV introduction event before preventive procedures were implemented, yielding an incidence rate of 2.41 per 100 farm-weeks. Most (63/100) of them occurred in sites where animals were placed negative and PEDV was detected in clinical samples in a median of 8 weeks post placement. After preventive procedures were implemented, the overall PEDV incidence rate dropped to 0.37 per 100 farm-weeks (84.65% reduction, p < 0.001). CONCLUSION: These results highlight the importance of systematic surveillance to identify the burden of diseases, areas of improvement in prevention and control, and to allow the measurement of the impact of policy/protocol changes.

Porcine reproductive and respiratory syndrome virus 2 (PRRSV-2) genetic diversity and occurrence of wild type and vaccine-like strains in the United States swine industry.

Porcine reproductive and respiratory syndrome virus genotype 2 (PRRSV-2) genetic diversity in the U.S. was assessed using a database comprising 10 years' worth of sequence data obtained from swine production systems routine monitoring and outbreak investigations. A total of 26,831 ORF5 PRRSV-2 sequences from 34 production systems were included in this analysis. Within group mean genetic distance (i.e. mean proportion of nucleotide differences within ORF5) per year according to herd type was calculated for all PRRSV-2 sequences. The percent nucleotide difference between each sequence and the ORF5 sequences from four commercially available PRRSV-2 vaccines (Ingelvac PRRS MLV, Ingelvac PRRS ATP, Fostera PRRS, and Prevacent PRRS) within the same lineage over time was used to classify sequences in wild-type or vaccine-like. The mean ORF5 genetic distance fluctuated from 0.09 to 0.13, being generally smaller in years in which there was a relative higher frequency of dominant lineage. Vaccine-like sequences comprised about one fourth of sequences obtained through routine monitoring of PRRS. We found that lineage 5 sequences were mostly Ingelvac PRRS MLV-like. Lineage 8 sequences up to 2011 were 62.9% Ingelvac PRRS ATP-like while the remaining were wild-type viruses. From 2012 onwards, 51.9% of lineage 8 sequences were Ingelvac PRRS ATP-like, 45.0% were Fostera PRRS-like, and only 3.2% were wild-type. For lineage 1 sequences, 0.1% and 1.7% of the sequences were Prevacent PRRS-like in 2009-2018 and 2019, respectively. These results suggest that repeated introductions of vaccine-like viruses through use of modified live vaccines might decrease within-lineage viral diversity as vaccine-like strains become more prevalent. Overall, this compilation of private data from routine monitoring provides valuable information on PRRSV viral diversity.

Phylogenetically Distinct Near-Complete Genome Sequences of Porcine Reproductive and Respiratory Syndrome Virus Type 2 Variants from Four Distinct Disease Outbreaks at U.S. Swine Farms over the Past 6 Years.

Porcine reproductive and respiratory syndrome virus (PRRSV) continues to mutate, causing disruptive PRRS outbreaks in farms that lead to reproductive failure and respiratory disease-associated mortality. We present four new PRRSV type 2 variants in the United States belonging to four distinct orf5 sublineages within lineage 1.

Forecasting viral disease outbreaks at the farm-level for commercial sow farms in the U.S.

Porcine epidemic diarrhea virus (PEDv) was introduced to the U.S. in 2013 and is now considered to be endemic. Like many endemic diseases, it is challenging for producers to estimate and respond to spatial and temporal variation in risk. Utilizing a regional spatio-temporal dataset containing weekly PEDv infection status for ∼15 % of the U.S. sow herd, we present a machine learning platform developed to forecast the probability of PEDv infection in sow farms in the U.S. Participating stakeholders (swine production companies) in a swine-dense region of the U.S. shared weekly information on a) PEDv status of farms and b) animal movements for the past week and scheduled movements for the upcoming week. Environmental (average temperature, humidity, among others) and land use characteristics (hog density, proportion of area with different land uses) in a 5 km radius around each farm were summarized. Using the Extreme Gradient Boosting (XGBoost) machine learning model with Synthetic Minority Over-sampling Technique (SMOTE), we developed a near real-time tool that generates weekly PEDv predictions (pertaining to two-weeks in advance) to farms of participating stakeholders. Based on retrospective data collected between 2014 and 2017, the sensitivity, specificity, positive and negative predictive values of our model were 19.9, 99.9, 70.5 and 99.4 %, respectively. Overall accuracy was 99.3 %, although this metric is heavily biased by imbalance in the data (less than 0.7 % of farms had an outbreak each week). This platform has been used to deliver weekly real-time forecasts since December 2019. The forecast platform has a built-in feature to re-train the predictive model in order to remain as relevant as possible to current epidemiological situations, or to expand to a different disease. These dynamic forecasts, which account for recent animal movements, present disease distribution, and environmental factors, will promote data-informed and targeted disease management and prevention within the U.S. swine industry.

Impact of nurse sows on influenza A virus transmission in pigs under field conditions.

Piglets prior to weaning play a central role in maintaining influenza infections in breeding herds and the use of nurse sows is a common practice to adopt piglets that fall behind and that otherwise would die. Transmission of influenza A virus (IAV) from nurse sows to adopted pigs has been reported experimentally, however, the importance of this route of transmission under field conditions has not yet been elucidated. A cohort study to assess the IAV status in nurse and control sows and their respective litters was carried out in three influenza positive breed-to-wean farms. A total of 94 control and 90 nurse sows were sampled by collecting udder skin wipes and oral swabs at enrollment (∼ 5-7 days after farrowing) and at weaning. Six piglets per litter were sampled randomly at enrollment, 2 days post-enrollment (DPE), 4 DPE, at day 14 of lactation (14DL) and at weaning. At enrollment, 76 % (69/91) of udder wipes and 3 % (3/89) of oral swabs from nurse sows were positive by rRT-PCR compared with 23 % (21/92) of udder wipes and 0 % (0/85) of oral swabs from control sows. Of the 94 control litters sampled, 11.7 %, 14.9 %, 22.9 %, 46.8 % and 63.9 % tested rRT-PCR IAV positive at enrollment, 2DPE, 4DPE, 14 DL and weaning, respectively. Corresponding prevalence for nurse sow litters were 12.2 %, 30.2 %, 37.0 %, 59.4 % and 56.4 %. The odds of IAV positivity were significantly higher (p < 0.05) for litters from nurse sows 2 DPE (odd ratio (OR) = 6.13, 95 % CI = 1.8-21.2), 4 DPE (OR = 5.5, 95 % CI = 1.7-17.8) and 14 DL (OR = 3.7, 95 % CI = 1.1-12.3). However, there were no differences in the proportion of positive samples at weaning. Moreover, approximately 18 % of the control sows and 11 % of nurse sows that tested IAV negative in oral swabs at enrollment, tested IAV positive at weaning. This study indicates that nurse sows can contribute to the transmission and perpetuation of IAV infections in pigs prior to weaning, particularly during the first week after adoption.

Temporal dynamics and the influence of environmental variables on the prevalence of avian influenza virus in main wetlands in central Chile.

Although wild birds are considered the main reservoir of the influenza A virus (IAV) in nature, empirical investigations exploring the interaction between the IAV prevalence in these populations and environmental drivers remain scarce. Chile has a coastline of more than 4000 kilometres with hundreds of wetlands, which are important habitats for both resident and inter-hemispheric migratory species. The aim of this study was to characterize the temporal dynamics of IAV in main wetlands in central Chile and to assess the influence of environmental variables on AIV prevalence. For that purpose, four wetlands were studied from September 2015 to June 2018. Fresh faecal samples of wild birds were collected for IAV detection by real-time RT-PCR. Furthermore, a count of wild birds present at the site was performed and environmental variables, such as temperature, rainfall, vegetation coverage (Normalized Difference Vegetation Index (NDVI)) and water body size, were determined. A generalized linear mixed model was built to assess the association between IAV prevalence and explanatory variables. An overall prevalence of 4.28% ± 0.28% was detected with important fluctuations among seasons, being greater during summer (OR = 4.87, 95% CI 2.11 to 11.21) and fall (OR = 2.59, 95% CI 1.12 to 5.97). Prevalence was positively associated with minimum temperature for the month of sampling and negatively associated with water body size measured two months before sampling, and NDVI measured three months before sampling. These results contribute to the understanding of IAV ecological drivers in Chilean wetlands providing important considerations for the global surveillance of IAV.

Assessing the litter level agreement of RT-PCR results for porcine reproductive and respiratory syndrome virus in testicles, tails and udder wipes diagnostic samples relative to serum from piglets.

Porcine reproductive and respiratory syndrome (PRRS) is currently the most detrimental disease in the U.S swine industry. Clinical signs of PRRS virus (PRRSv) infection in breeding herds include reproductive failure with abortions, stillbirths, premature farrowings and increased pre-weaning mortality. Serum from due-to-wean piglets is considered the most suitable specimen to monitor PRRSv infection and stability in breeding herds. However, processing fluids (PF - the serosanguinous exudate resultant of the collection of tails and testicles during processing) are a new specimen proposed to monitor piglets at processing (3-5 days of age) and udder wipes (UW) of lactating sows is yet another specimen to monitor infection status of suckling piglets indirectly. Here, we assessed which specimen type (e.g. sera, testicles, tails or UW) should be used to accurately establish the PRRSv status of a litter. Twenty-four litters were conveniently selected on a farm at 10 weeks post PRRSv outbreak. Blood samples, tails and testicles from every piglet in a litter, and an udder skin wipe from the sow were collected at processing (3-5 days). Individual litter testicles and tails as well as the udder wipe were placed each in a reclosable bag to prevent cross-contamination. Sensitivity (Se), specificity (Sp), negative predictive value (NPV), positive predictive value (PPV) and global agreement at the litter level were calculated using the sera results of the litter as the gold standard. The optimum cycle threshold (Ct) value to classify a sample as negative was ≥35 for serum and ≥36 for the aggregated samples (testicles, tails, and UW) based on the ROC curve analysis. Using those thresholds, the fluid collected from the testicles showed the best overall performance (Se = 92 % [62-100]; Sp = 82 % [48-98], NPV = 90 % [55-100], PPV = 85 % [55-98], global agreement = 87 %) compared to tail fluid and UW. Sensitivity of the tail fluid was 62 % (32-86) and the UW was 23 % (5-54), both of which yielded a 100 % specificity and PPV. This study provides information on the contribution of each of the tissues collected at processing on the detection of PRRSv, which becomes relevant in countries were castration and/or tail docking is banned.

Spatial relative risk and factors associated with porcine reproductive and respiratory syndrome outbreaks in United States breeding herds.

Details of incident cases of porcine reproductive and respiratory syndrome (PRRS) in United States breeding herds were obtained from the Morrison's Swine Health Monitoring Project. Herds were classified as cases if they reported an outbreak in a given season of the year and non-cases if they reported it in a season other than the case season or if they did not report a PRRS outbreak in any season. The geographic distribution of cases and non-cases was compared in each season of the year. The density of farms that had a PRRS outbreak during summer was higher in Southern Minnesota and Northwest-central Iowa compared to the density of the underlying population of non-case farms. This does not mean that PRRS outbreaks are more frequent during summer in absolute terms, but that there was a geographical clustering of herds breaking during summer in this area. Similar findings were observed in autumn. In addition, the density of farms reporting spring outbreaks was higher in the Southeast of the United States compared to that of the underlying population of non-case farms. A similar geographical clustering of PRRS outbreaks was observed during winter in the Southeast of the United States. Multivariable analyses, adjusting for the effect of known confounders, showed that the incidence rate of PRRS was significantly lower during winter and autumn during the porcine epidemic diarrhea (PED) epidemic years (2013-2014), compared to PRRS incidence rates observed during the winter and autumn of PED pre-epidemic years (2009-2012). After 2014, an increase in the incidence rate of PRRS was observed during winter and spring but not during autumn or summer. Pig dense areas were associated with a higher incidence rate throughout the year. However, this association tended to be stronger during the summer. Additionally, herds with ≥2500 sows had an increased incidence rate during all seasons except spring compared to those with <2500 sows. PRRS incidence was lower in year-round air-filtered herds compared to non-filtered herds throughout the year. We showed that not only the spatial risk of PRRS varies regionally according to the season of the year, but also that the effect of swine density, herd size and air filtering on PRRS incidence may also vary according to the season of the year. Further studies should investigate regional and seasonal drivers of disease. Breeding herds should maintain high biosecurity standards throughout the year.

Risk factors and spatial relative risk assessment for influenza A virus in poultry and swine in backyard production systems of central Chile.

Backyard production systems (BPS) are a common form of poultry and swine production worldwide. The limited implementation of biosecurity standards in these operations makes BPS a potential source for the emergence of pathogens that have an impact on both animal and public health. Information regarding circulation of influenza A virus (IAV) in poultry and swine raised in BPS is scarce; particularly in South American countries. The objective of this study was to estimate prevalence and seroprevalence of IAV in BPS in central Chile, identify subtype diversity, evaluate risk factors and spatial relative risk for IAV. Samples were collected from 329 BPS from central Chile. Seroprevalence at BPS level was 34.7% (95% CI: 23.1%-46.2%), 19.7% (95% CI: 9.9%-30.6%) and 11.7% (95% CI: 7.2%-16.4%), whereas prevalence at BPS level was 4.2% (95% CI: 0.0%-8.8%), 8.2% (95% CI: 0.8%-14.0%) and 9.2% (95% CI: 4.8%-13.1%), for the Metropolitan, Valparaiso and LGB O'Higgins regions, respectively. Spatial analysis revealed that central-western area of Metropolitan region and the southern province of Valparaiso region could be considered as high-risk areas for IAV (spatial relative risk = 2.2, p < .05). Logistic regression models identified the practice of breeding both poultry and pigs at the BPS as a risk factor (95% CI 1.06-3.75). From 75 IAV ELISA-positive sera, 20 chicken sera had haemagglutination inhibition titres ranging from 20 to 160, and of these, 11 had microneutralization titres ranging from 40 to 960 for one or more IAV subtypes. Identified subtypes were H1, H3, H4, H9, H10 and H12. Results from this study highlight the need for further IAV surveillance programmes in BPS in Chile. Early detection of IAV strains circulating in backyard animals, especially in regions with large human populations, could have an enormous impact on animal and public health.

Estimation of the burden of leptospirosis in New Zealand.

BACKGROUND: Human leptospirosis mainly affects people in close occupational contact with domestic livestock and their products in New Zealand. The disease has an unquantified impact on both human health and animal production in the country. This study aimed to estimate the burden of leptospirosis in terms of disability-adjusted life years (DALYs) and cost associated with loss due to absence from work, treatment of disease, animal production loss and cost of vaccination. METHODS: Previously published studies of abattoir workers farmers, and veterinarians, reporting annual risks of influenza-like illness attributable to Leptospira infection, were used to estimate the expected number of cases in a year. The cost of lost animal production was based on results of observational studies in beef cattle, sheep and deer conducted in New Zealand. RESULTS: Expected median annual number of severe and mild cases of human leptospirosis was 2,025 (95% probability interval [95% PI] 1,138-3,422). Median annual DALYs were 0.42 (95% PI: 0.06-2.40) per 100,000 people for the entire population, and 15.82 (95% PI: 2.09-90.80) per 100,000 people working in at-risk occupations (i.e. abattoir workers, farmers and veterinarians). Human infection resulted in a median cost of 4.42 (95% PI: 2.04-8.62) million US dollars (USD) due to absence from work and disease treatment. Median production loss cost in beef cattle, sheep and deer was USD 7.92 (95% PI: 3.75-15.48) million, while median vaccination cost in cattle, (including dairy), sheep and deer was USD 6.15 (95% PI: 5.30-7.03) million. Total annual cost of leptospirosis plus vaccination was USD 18.80 (95% PI: 13.47-27.15) million, equivalent to USD 440,000 (95% PI: 320,000-640,000) per 100,000 people. CONCLUSION: This study provides an estimate of the disease burden and cost of leptospirosis in New Zealand that could support occupational health authorities and livestock industries in assessing interventions for this disease.

Temporal Dynamics of Co-circulating Lineages of Porcine Reproductive and Respiratory Syndrome Virus.

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is the most important endemic pathogen in the U.S. swine industry. Despite control efforts involving improved biosecurity and different vaccination protocols, the virus continues to circulate and evolve. One of the foremost challenges in its control is high levels of genetic and antigenic diversity. Here, we quantify the co-circulation, emergence and sequential turnover of multiple PRRSV lineages in a single swine-producing region in the United States over a span of 9 years (2009-2017). By classifying over 4,000 PRRSV sequences (open-reading frame 5) into phylogenetic lineages and sub-lineages, we document the ongoing diversification and temporal dynamics of the PRRSV population, including the rapid emergence of a novel sub-lineage that appeared to be absent globally pre-2008. In addition, lineage 9 was the most prevalent lineage from 2009 to 2010, but its occurrence fell to 0.5% of all sequences identified per year after 2014, coinciding with the emergence or re-emergence of lineage 1 as the dominant lineage. The sequential dominance of different lineages, as well as three different sub-lineages within lineage 1, is consistent with the immune-mediated selection hypothesis for the sequential turnover in the dominant lineage. As host populations build immunity through natural infection or vaccination toward the most common variant, this dominant (sub-) lineage may be replaced by an emerging variant to which the population is more susceptible. An analysis of patterns of non- synonymous and synonymous mutations revealed evidence of positive selection on immunologically important regions of the genome, further supporting the potential that immune-mediated selection shapes the evolutionary and epidemiological dynamics for this virus. This has important implications for patterns of emergence and re-emergence of genetic variants of PRRSV that have negative impacts on the swine industry. Constant surveillance on PRRSV occurrence is crucial to a better understanding of the epidemiological and evolutionary dynamics of co-circulating viral lineages. Further studies utilizing whole genome sequencing and exploring the extent of cross-immunity between heterologous PRRS viruses could shed further light on PRRSV immunological response and aid in developing strategies that might be able to diminish disease impact.

Indirect assessment of porcine reproductive and respiratory syndrome virus status in pigs prior to weaning by sampling sows and the environment.

There is a need to develop cost effective approaches to sample large populations in particular to determine the disease status of pigs prior to weaning. In this study we assessed the presence of the porcine reproductive and respiratory syndrome virus (PRRSV) in the environment (surfaces and air) of farrowing rooms, and udder skin of lactating sows as an indirect measure of piglet PRRSV status. Samples were collected at processing and weaning every three weeks for 23 weeks after a PRRSV outbreak was diagnosed in a swine breeding herd. PRRSV was detected at processing in udder skin wipes, environmental wipes and airborne deposited particle samples up to 14 weeks post outbreak and at weaning in udder skin wipes up to 17 weeks post outbreak. Similar sensitivities were observed for udder skin wipes (43% [95% CI: 23%-66%]) and surface wipes (57% [95% CI: 34%-77%]) when compared to serum at the litter level from piglets at processing. PRRSV was detected in the environment and the udder skin of lactating sows, which indicates that aggregate samples of the environment or lactating sows may be used to evaluate the PRRSV status of the herd in pigs prior to weaning. However, the use of environmental samples to detect PRRSV by RT-PCR should not be used as the single method to assess the PRRSV status at the litter level. Furthermore, our findings also highlight potential sources of PRRSV infection for piglets in breeding herds.

Detection of Mycoplasma hyopneumoniae in piglet processing fluids.

Processing fluid (PF) is a sample type composed of fluids obtained from testicles and tails as the product of piglet processing. Mycoplasma hyopneumoniae is a bacterium that colonises the respiratory tract of pigs and has rarely been detected in tissues outside the respiratory system. No data exist in the literature regarding detection of M hyopneumoniae in PF or its use for herd monitoring of this pathogen. The main goal of this study was to evaluate the feasibility of detecting M hyopneumoniae in PF. Testicles and tails of 21 conveniently selected litters from a commercial sow farm were collected, by litter, and tested for M hyopneumoniae by real time-PCR. Daily aggregated processing tissues were collected for a two-month period to assess the detection of M hyopneumoniae in PF. The comparison in the percentage of positive samples in fluids from testicles (38 per cent, 8/21) or tails (4.8 per cent, 1/21) was significantly different (P=0.023). The percentage of daily aggregated PF with cycle threshold values up to 37 was 52.9 per cent (9/17) and 26.7 per cent (4/15) for December and January, respectively. Overall, these data show detection of M hyopneumoniae in PF for the first time and points at the potential use of this sample for monitoring of this bacterium in breeding farms.

Aerosol Detection and Transmission of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV): What Is the Evidence, and What Are the Knowledge Gaps?

In human and veterinary medicine, there have been multiple reports of pathogens being airborne under experimental and field conditions, highlighting the importance of this transmission route. These studies shed light on different aspects related to airborne transmission such as the capability of pathogens becoming airborne, the ability of pathogens to remain infectious while airborne, the role played by environmental conditions in pathogen dissemination, and pathogen strain as an interfering factor in airborne transmission. Data showing that airborne pathogens originating from an infectious individual or population can infect susceptible hosts are scarce, especially under field conditions. Furthermore, even though disease outbreak investigations have generated important information identifying potential ports of entry of pathogens into populations, these investigations do not necessarily yield clear answers on mechanisms by which pathogens have been introduced into populations. In swine, the aerosol transmission route gained popularity during the late 1990's as suspicions of airborne transmission of porcine reproductive and respiratory syndrome virus (PRRSV) were growing. Several studies were conducted within the last 15 years contributing to the understanding of this transmission route; however, questions still remain. This paper reviews the current knowledge and identifies knowledge gaps related to PRRSV airborne transmission.