Farm management practices associated with influenza A virus contamination of people working in Midwestern United States swine farms.

Indirect transmission of influenza A virus (IAV) contributes to virus spread in pigs. To identify farm management activities with the ability to contaminate farmworkers' hands and clothing that then could be a source of virus spread to other pigs, we conducted a within-farm, prospective IAV surveillance study. Hands and clothes from farmworkers performing the activities of piglet processing, vaccination, or weaning were sampled before and after the activities were performed. Samples were tested by IAV rRT-PCR and virus viability was assessed by cell culture. A multivariate generalized linear model was used to detect associations of the activities with IAV contamination. Of the samples collected for IAV rRT-PCR testing, there were 16% (12/76) collected immediately after processing, 96% (45/48) collected after vaccination, and 94% (29/31) collected after weaning that tested positive. Samples collected immediately after vaccination and weaning, i.e., activities that took place during the peri-weaning period when pigs were about 3 weeks of age, had almost 6 times higher risk of IAV detection and had more samples IAV positive (p-value < 0.0001) than samples collected after processing, i.e., an activity that took place in the first few days of life. Both, hands and clothes had similar contamination rates (46% and 55% respectively, p-value = 0.42) and viable virus was isolated from both. Our results indicate that activities that involve the handling of infected piglets close to weaning age represent a significant risk for IAV dissemination due to the high level of IAV contamination found in farmworkers' hands and coveralls involved in the activities. Biosecurity protocols that include hand sanitation and changing clothing after performing activities with a high-risk of influenza contamination should be recommended to farmworkers to control and limit the mechanical spread of IAV between pigs.

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.

Genetic variability of influenza A virus in pigs at weaning in Midwestern United States swine farms.

Suckling piglets play an important role at maintaining influenza A virus (IAV) infections in breeding herds and disseminating them to other farms at weaning. However, the role they play at weaning to support and promote genetic variability of IAV is not fully understood. The objective here was to evaluate the genetic diversity of IAV in pigs at weaning in farms located in the Midwestern USA. Nasal swabs (n = 9,090) collected from piglets in breed-to-wean farms (n = 52) over a six-month period across seasons were evaluated for the presence of IAV. Nasal swabs (n = 391) from 23 IAV-positive farms were whole-genome sequenced. Multiple lineages of HA (n = 7) and NA (n = 3) were identified in 96% (22/23) and 61% (237/391) of the investigated farms and individual piglets, respectively. Co-circulation of multiple types of functional HA and NA was identified in most (83%) farms. Whole IAV genomes were completed for 126 individual piglet samples and 25 distinct and 23 mixed genotypes were identified, highlighting significant genetic variability of IAV in piglets. Co-circulation of IAV in the farms and co-infection of individual piglets at weaning was observed at multiple time points over the investigation period and appears to be common in the investigated farms. Statistically significant genetic variability was estimated within and between farms by AMOVA, and varying levels of diversity between farms were detected using the Shannon-Weiner Index. Results reported here demonstrate previously unreported levels of molecular complexity and genetic variability among IAV at the farm and piglet levels at weaning. Movement of such piglets infected at weaning may result in emergence of new strains and maintenance of endemic IAV infection in the US swine herds. Results presented here highlight the need for developing and implementing novel, effective strategies to prevent or control the introduction and transmission of IAV within and between farms in the country.

Effect of influenza A virus sow vaccination on infection in pigs at weaning: A prospective longitudinal study.

Although vaccination is the main measure to control influenza A virus (IAV) in swine, there is limited information on the efficacy of sow vaccination on reducing IAV infections in pigs at weaning. We assessed the effect of sow vaccination on IAV infection in pigs at weaning in a cohort of 52 breeding herds studied prospectively. Herds were voluntarily enrolled according to their IAV history, sow vaccination protocol and monitored during six months (prospective longitudinal study). On each herd, nasal swabs were collected monthly from 30 pigs at weaning and tested for IAV by RT-PCR. IAV was detected in 25% (75/305) of sampling events. Of 9,150 nasal swab pools (3 individual nasal swabs/pool), 15% (458/3050) of pools tested IAV positive. IAV infections in pigs at weaning were lower in vaccinated herds compared to non-vaccinated ones. Moreover, no significant differences were seen between prefarrow and whole herd protocols, or the use of commercial versus autogenous IAV vaccines. Prefarrow and whole herd vaccination protocols reduced the odds of groups testing IAV positive at weaning in comparison with no vaccination. Our results are relevant when considering implementation of sow vaccination to control influenza infections in pigs at weaning and, hence, minimize transmission to growing pigs and other farms.

Relationship between weaning age and antibiotic usage on pig growth performance and mortality.

A total of 2,184 pigs (DNA 600 × PIC L42) were used to evaluate the effects of weaning age and antibiotic (AB) use on pig performance from weaning to marketing in a commercial production system. Experimental treatments were arranged in a 3 × 2 factorial with main effects of weaning age (18.5, 21.5, or 24.5 d of age) and with the use of ABs or an antibiotic-free (NAE) program. At birth, pigs were ear tagged, and the date of birth and sex recorded. Pigs were weaned from a 4,000-sow farm over four consecutive weeks. Four weaning batches (one per week) of 546 pigs were used. Each weaning batch had one-third of pigs of each weaning age. Pigs were placed in pens by weaning age and then randomly assigned to an AB or NAE program. There were 14 replicate pens per treatment and 26 pigs per pen (13 barrows and 13 gilts). Pigs allocated to the AB program were fed a diet containing 441 mg/kg chlortetracycline (CTC) from day 8 to 21 postweaning. They were also administered 22 mg/kg of body weight (BW) of CTC via drinking water for five consecutive days after a porcine respiratory and reproductive syndrome outbreak during week 7 after weaning. In the first 42 d postweaning, increasing weaning age improved (linear, P < 0.001) BW at day 42, average daily gain (ADG), and average daily feed intake (ADFI). From weaning to 197 d of age, increasing weaning age increased (linear, P < 0.001) ADG and ADFI. Pigs on the AB program had greater (P = 0.031) ADG and ADFI compared with NAE pigs. An interaction (linear, P = 0.005) was observed for feed efficiency (G:F). When ABs were provided, increasing weaning age did not result in any change in G:F; however, in the NAE program, increasing weaning age increased G:F. Pigs on the AB program had lower (P < 0.001) total losses (mortality and removals) than those on the NAE program. Increasing weaning age marginally (linear, P = 0.097) decreased total losses. Increasing weaning age decreased (quadratic, P < 0.001) the number of pigs treated with an injectable AB but the AB program did not (P = 0.238). The weight sold (at 197 d of age) per pig weaned was increased (linear, P = 0.050) by increasing weaning age and by using AB in feed and water (P = 0.019). In summary, increasing weaning age linearly improved most of the pig performance criteria and relatively the short-term use of ABs reduced mortality and removals with both factors contributing to increased weight sold per pig weaned.

Effect of strain-specific maternally-derived antibodies on influenza A virus infection dynamics in nursery pigs.

Reducing the number of influenza A virus (IAV) infected pigs at weaning is critical to minimize IAV spread to other farms. Sow vaccination is a common measure to reduce influenza levels at weaning. However, the impact of maternally-derived antibodies on IAV infection dynamics in growing pigs is poorly understood. We evaluated the effect of maternally-derived antibodies at weaning on IAV prevalence at weaning, time of influenza infection, number of weeks that pigs tested IAV positive, and estimated quantity of IAV in nursery pigs. We evaluated 301 pigs within 10 cohorts for their influenza serological (seroprevalence estimated by hemagglutination inhibition (HI) test) and virological (prevalence) status. Nasal swabs were collected weekly and pigs were bled 3 times throughout the nursery period. There was significant variability in influenza seroprevalence, HI titers and influenza prevalence after weaning. Increase in influenza seroprevalence at weaning was associated with low influenza prevalence at weaning and delayed time to IAV infection throughout the nursery. Piglets with IAV HI titers of 40 or higher at weaning were also less likely to test IAV positive at weaning, took longer to become infected, tested IAV RT-PCR positive for fewer weeks, and had higher IAV RT-PCR cycle threshold values compared to piglets with HI titers less than 40. Our findings suggest that sow vaccination or infection status that results in high levels of IAV strain-specific maternally-derived antibodies may help to reduce IAV circulation in both suckling and nursery pigs.

Breed-to-wean farm factors associated with influenza A virus infection in piglets at weaning.

Breed-to-wean pig farms play an important role in spreading influenza A virus (IAV) because suckling piglets maintain, diversify and transmit IAV at weaning to other farms. Understanding the nature and extent of which farm factors drive IAV infection in piglets is a prerequisite to reduce the burden of influenza in swine. We evaluated the association between IAV infection in piglets at weaning and farm factors including farm features, herd management practices and gilt- and piglet-specific management procedures performed at the farm. Voluntarily enrolled breed-to-wean farms (n = 83) agreed to share IAV diagnostic testing and farm data from July 2011 through March 2017 including data obtained via the administration of a survey. There were 23% IAV RT-PCR positive samples of the 12,814 samples submitted for IAV testing within 2989 diagnostic submissions with 30% positive submissions. Among all the factors evaluated (n = 24), and considering the season-adjusted multivariable analysis, only sow IAV vaccination and gilt IAV status at entry significantly reduced (p-value<0.05) IAV infections in piglets at weaning. Results from this study indicate that veterinarians and producers could manage these identified factors to reduce the burden of influenza in piglets prior to wean and perhaps, reduce the spread of IAV to other farms and people.

Antigenic drift of H1N1 influenza A virus in pigs with and without passive immunity.

BACKGROUND: The genetic and antigenic characteristics of influenza A viruses (IAV) within and between species change over time due to antigenic shift and drift. Although pigs are known to play a key role in the epidemiology of IAV between species, little is known about the molecular evolution of IAV hemagglutinin (HA) in pigs. OBJECTIVES: The aim of this study was to evaluate the HA drift of an H1N1 IAV after infecting weaned pigs with or without maternally derived passive immunity. METHODS: Three- to four-week-old piglets born either to vaccinated or unvaccinated sows were contact-infected upon exposure with an IAV-infected pig. Nasal swabs were collected daily from each pig and tested for IAV by RRT-PCR. Full-length HA sequences were obtained directly from positive nasal swabs and compared between groups. RESULTS: Synonymous and non-synonymous mutations were detected in pigs with and without passive immunity. Most of the non-synonymous mutations occurred within the HA1 region of the HA. Changes within HA1 region were only identified in antigenic site B in pigs without passive immunity and in antigenic sites A, B, and D in pigs with passive immunity. However, there was no association between the immune status of the pig and the amino acid substitutions observed. CONCLUSIONS: Overall, we demonstrated that amino acid substitutions within antigenic sites can happen in weaned pigs with or without passive immunity shortly after infection.