In this final episode, Dr. Montserrat Torremorell (University of Minnesota), Dr. Amy Vincent (USDA Agricultural Research Service), Dr. Christa Goodell (Boehringer Ingelheim), Dr. Gordon Spronk (Pipestone Veterinary Services), and Dr. Tom Wetzell (Boehringer Ingelheim) discuss the control of IAV-S, the research & development of vaccines for IAV-S, and the products that are being developed from the research.
In this second episode, Dr. Montserrat Torremorell, Dr. Adam Schelkopf (Pipestone Veterinary Services), Dr. Gordon Spronk (Pipestone Veterinary Services), and Dr. Tom Wetzell (Boehringer Ingelheim) continue the conversation on the challenges of IAV-S in day to day operation, the approaches to identifying infected pigs, and the processes that need to be put in place to reduce infection and increase survivability of pigs.
Podcasts are perfect for summer! We are presenting you with a new series on swine influenza from “At The Meeting… Honoring Dr. Bob Morrison in collaboration with SwineCast.
In this first episode, Dr. Montserrat Torremorell (University of Minnesota), Dr. Marie Culhane (University of Minnesota), Dr. Gordon Spronk (Pipestone Veterinary Services), and Dr. Tom Wetzell (Boehringer Ingelheim), talk about the issues of influenza in humans and swine, the state of surveillance of influenza in pigs and humans, and the biosecurity needed to help prevent the spread of the influenza virus between human and pigs.
Today we are sharing a publication from the Torremorell lab regarding the impact of vaccination (both homologous and heterologous) on the detection of swine influenza virus in aerosols. The full publication is available in open access online on the PlosOne website.
Influenza A virus can be transmitted by direct and indirect contact and aerosols. Indeed, the virus has been detected and isolated from aerosols generated from pigs with and without immunity. Since then, there has been increased evidence of the role of aerosols in influenza transmission among swine.
Vaccination is used in swine populations as a strategy to mitigate clinical effects and the economic impact of influenza infections. It has also been proven to reduce shedding in pigs. Additionally, a study on the transmission of influenza in ferrets showed that high temperature may decrease the risk of airborne transmission. Therefore, we wondered if combining vaccination and high temperature would affect the detection of influenza virus in the air.
The objective of this study was to assess the effect of vaccination on the generation of influenza A virus bioaerosols under warm conditions in pigs with varying degrees of cross-protective immunity.
Material and Methods
36 pigs of three weeks of age, seronegative for influenza were separated into four groups:
vaccinated with an influenza strain identical to the one used for the challenge (homologous)
vaccinated with a commercial vaccine containing multiple strains of influenza, all different from the challenge strain (heterologous, multivalent)
vaccinated with a commercial vaccine containing one influenza strain different from the challenge strain (heterologous, monovalent)
unvaccinated, which received an injection of saline instead
Pigs were challenged intranasally and intratracheally with a strain of H1N1 influenza virus, two weeks after the last vaccination.
Serum collected the day prior to the vaccination and at the end of the study 14 days post inoculation were tested via hemagglutination inhibition (HI) and ELISA.. Nasal swabs and oral fluids were collected and tested via PCR. Air samples were collected three times a day and tested via PCR and virus isolation. Temperature and humidity were recorded every five minutes.
Hemagglutination inhibition and ELISA
Prior to infection, pigs in group 1 (Vaccinated, homologous) had significantly higher HI titers compared to the other three groups. In the group 3 (vaccinated, heterologous monvalent) 4 pigs had HI titers against the challenge strain, while pigs in groups 2 and 4 were negative against the challenge strain. All groups were HI positive against the challenge strain at necropsy, however HI titers were statistically different between group 4 and groups 1 and 3.
Proportion of pigs infected
The proportion of pigs infected was significantly higher in group 4 than in the vaccinated ones. Also, the percentage of infected pigs in group 1 was significantly lower than in group 2, but there was no difference with group 3.
Nasal swabs and oral fluids
Pigs in group 4 had higher amounts of nasal virus shedding most of the sampling days compared to vaccinated groups. Additionally, group 2 had higher levels of IAV compared with groups 1 and 4. Oral fluid results were in agreement with nasal swab.
All air samples in the vaccinated groups tested negative by RRT-PCR. Air samples collected at days 1, 2 and 3 from NON-VAC pigs tested positive by RRT-PCR but negative by virus isolation
The 2009 influenza pandemic, the variant H3N2v viruses in agricultural fairs and the zoonotic poultry H5N9 infections in China have highlighted the constant threat that influenza A viruses (IAV) present to people and animals. In this study we evaluated the effect of IAV vaccination on aerosol shedding in pigs housed in warm environmental conditions. Thirty-six, three-week old weaned pigs were obtained from an IAV negative herd and were randomly allocated to one of 4 groups: 1) a homologous vaccine group, 2) a heterologous multivalent vaccine group, 3) a heterologous monovalent group and, 4) a non-vaccinated group. After vaccination pigs were challenged with the triple reassortant A/Sw/IA/00239/04 H1N1 virus. Environmental temperature and relative humidity were recorded throughout the study. Nasal swabs, oral fluids and air samples were collected daily. All samples were tested by RRT-PCR and virus isolation was attempted on positive samples. Average temperature and relative humidity throughout the study were 27°C (80°F) and 53%, respectively. A significantly higher proportion of infected pigs was detected in the non-vaccinated than in the vaccinated group. Lower levels of nasal virus shedding were found in vaccinated groups compared to non-vaccinated group and IAV was not detected in air samples of any of the vaccinated groups. In contrast, positive air samples were detected in the non-vaccinated group at 1, 2 and 3 days post infection although the overall levels were considered low most likely due to the elevated environmental temperature. In conclusion, both the decrease in shedding and the increase in environmental temperature may have contributed to the inability to detect airborne IAV in vaccinated pigs.
Pooling oral fluid samples seems to be a good strategy to determine the status of a farm (positive/negative) for influenza A virus (IAV) and PRRSV.
Sampling water cups using environmental Swiffer™ samples appears to be a sensitive approach to detect IAV at the pen level.
However, sample size has been limited to one farm.
The objective of this project was to compare the sensitivity of pooled pen oral fluids (OF) and environmental samples (Swiffer™ kits on water cups) using individual pen oral fluids as the standard.
Fifteen paired environmental and individual pen OF were collected at days 3, 7, 10, 17, 24 and 31 post placement in two different nursery farms. Environmental samples (ES) were taken using Swiffer™ cloths to sample the bottom of water cups (both pans and bowls), focusing around nipples. After individual samples were collected, pen OF were pooled by 3.
There was an overall sensitivity of 71% (IAV) and 14% (PRRS) for the ES samples compared to individual OF. Pooled oral fluids samples had an overall sensitivity of 50%(IAV)and 80%(PRRSV)relative to individual pen OF.
In summary, ES appears to be a good strategy when sampling for IAV and not a reliable option when trying to diagnose PRRSV.
Earlier this year, Dr. Fabian Chamba Pardo successfully defended his PhD under the supervision of Drs. Montse Torremorell and Marie Culhane. The focus of his thesis is influenza epidemiology with an emphasis on sow farms and nurseries. We share with you today a summary of his work.
Influenza is an economically important disease in pigs and a public health threat. Breed-to-wean (BTW) farms play a central role in influenza epidemiology and control because piglets maintain and disseminate influenza A virus (IAV) to other farms. Despite the importance of piglets in influenza epidemiology, there is limited information on IAV infection parameters in piglets, risk factors that impact IAV prevalence in piglets at weaning, and how strategies that are implemented in BTW farms affect IAV infections in weaned pigs.
In this thesis, my goal was to address some of the questions that are central to the transmission and control of influenza in BTW farms, especially infection in piglets ready to wean. The questions addressed are also critical to guide control strategies to mitigate IAV infections in the post weaning period. More specifically, I aimed to: 1) estimate herd-level prevalence and seasonality of influenza in BTW farms, 2) evaluate farm factors associated with IAV infection in piglets at weaning, 3) assess transmission patterns and parameters of influenza in nursery pigs based on IAV prevalence at weaning, and 4) evaluate the impact of maternally-derived antibodies (MDA) at weaning on IAV infection parameters in nursery pigs.
Research Chapter 1
Influenza herd-level prevalence and seasonality in breed-to-wean pig farms in the Midwestern United States
Results showed that IAV herd-level prevalence in piglets at weaning from Midwestern BTW farms is seasonal with higher infection rates in winter (December) and spring (May) than those in summer and fall. Additionally, influenza seasonality was partially explained by the seasonal variations of outdoor air absolute humidity and temperature. Finally, there was significant genetic diversity of influenza strains circulating in those farms and that, co-circulation of more than one genetically distinct clade over time was very common in the studied farms. This is critical knowledge that may help to identify high risk periods where influenza control measures can be placed. It may also help to create research opportunities on absolute humidity and influenza transmission in pigs and finally, it supports other studies that have shown that genetic diversity and circulation is wide and common and that new vaccines and vaccination strategies should take that into consideration.
Research Chapter 2
Breed-to-wean farm factors associated with influenza A virus infection in piglets at weaning
In this chapter, there were 24 farm factors evaluated for their association with influenza at weaning and among those, only IAV sow vaccination and the IAV-negative status of replacement breeding females (gilts) at entry to the herd were significantly associated with less IAV infected piglets at weaning. This is critical information that veterinarians and producers may use to manage IAV levels at weaning. In addition, there was also a lack of significant association with factors such as air filtration and farm density which may be indicative that endemic influenza infections are more important than airborne lateral transmissions between farms. Finally, disease control strategies such as herd closure, early weaning, batch farrowing, gilt isolation and gilt influenza vaccination were not fully evaluated in this study. Hence, more work is needed to further understand how to use these strategies to decrease influenza infections in pigs.
Research Chapter 3
Influenza A virus transmission patterns and parameters in growing pigs
Results indicate that groups of piglets with different prevalence at weaning had different transmission patterns and parameters after weaning and these patterns were characterized by 1, 2 or no peaks of infection after weaning. Piglets with low prevalence at weaning had less influenza infections in the nursery. This information may help producers and veterinarians to make informed decisions when it comes to use control strategies such as sow vaccination aimed to reduce influenza infections in the nursery.
Research Chapter 4
Effect of maternally-derived antibodies on influenza A virus infection in growing pigs
In my last chapter, I reported that if pigs had high levels of strain-specific maternally-derived antibodies at weaning, IAV infection occurred later and it was of shorter duration after weaning. Piglets with hemagglutination inhibition (HI) titers of 1:40 or higher were less likely to test IAV positive at weaning and during the nursery. These results indicate that strain-specific maternally-derived antibodies generated with sow vaccination pre-farrow significantly reduce influenza infections at weaning and in the nursery.
Knowledge of influenza seasonality and what factors are significantly associated with influenza in breed-to-wean farms can help producers and veterinarians to better use and allocate influenza control strategies such as sow vaccination. In addition, lower prevalence of influenza at weaning due to high strain-specific maternally-derived antibodies levels may help decrease influenza spread from wean-to-finish farms. Reducing the burden of influenza in growing pigs should decrease influenza-associated economic losses and the generation of novel strains, including strains with pandemic potential. More studies are needed to further elucidate control strategies to limit influenza infections and spread in pigs.
Influenza is endemic and seasonal in piglets from sow farms in the Midwest with higher infections in winter and spring.
Influenza seasonality was partially explained by outdoor air absolute humidity and temperature trends.
Influenza genetic diversity was high and co-circulation of more than one genetically distinct virus was common.
To study influenza levels over time and its seasonality, monthly testing data of piglets at weaning from 34 sow farms during ~5 years were analyzed.
There were 28% of positive submissions with a median influenza herd-level prevalence of 28%. Genetic diversity was significant with 10 genetically distinct clades of contemporary US swine influenza viruses as shown below. Furthermore, 21% of farms had 3 genetically distinct viruses circulating over time; 18% had 2, 41% had 1 and 20% had no isolates available.
In summary, influenza herd-level prevalence in Midwestern sow farms had a seasonal pattern with higher levels in winter and spring. This is important to better allocate influenza control strategies such as vaccination in sow farms. Influenza seasonality was partially explained by outdoor air absolute humidity and temperature although other factors such as immunity and new introductions may play a role in the observed seasonality.