Antibodies of influenza A(H1N1)pdm09 virus in pigs' sera cross-react with other influenza A virus subtypes. A retrospective epidemiological interpretation of Norway's serosurveillance data from 2009-2017.

Since the incursion of influenza A(H1N1)pdm09 virus in 2009, serosurveillance every year of the Norwegian pig population revealed the herd prevalence for influenza A(H1N1)pdm09 (HIN1pdm09) has stabilised between 40% and 50%. Between 30 September 2009 and 14 September 2017, the Norwegian Veterinary Institute and Norwegian Food Safety Authority screened 35,551 pigs for antibodies to influenza A viruses (IAVs) from 8,636 herds and found 26% or 8,819 pigs' sera ELISA positive (titre ≥40). Subtyping these IAV antibodies from 8,214 pigs in 3,629 herds, by a routine haemagglutination inhibition test (HAIT) against four standard antigens produced 13,771 positive results (HAIT titre ≥40) of binding antibodies. The four antigen subtypes eliciting positive HAIT titre in descending frequencies were immunogen H1N1pdm09 (n = 8,200 or 99.8%), swine influenza A virus (SIVs) subtypes swH1N1 (n = 5,164 or 62%), swH1N2 (n = 395 or 5%) and swH3N2 (n = 12 or 0.1%). Of these 8,214 pig pigs sera, 3,039 produced homologous HAIT subtyping, almost exclusively immunogen H1N1pdm09 (n = 3,026 or 99.6%). Using HAIT titre of pig and herd geometric mean titre (GMT) as two continuous outcome variables, and with the data already structured hierarchically, we used mixed effects linear regression analysis to investigate the impact of predictors of interests had on the outcomes. For the full data, the predictors in the regression model include categorical predictors antigen subtype (H1N1pdm09, swH1N1, swH1N2 & swH3N2), and production type (sow herd or fattening herd), ordinal predictors year (longitudinally from 2009 to 2017) and number of antigens in heterologous reactions (1, 2, 3, 4) in the same pig serum. The last predictor, the proportion of HAIT positive (antigen specific) in tested pigs within the herd, was a continuous predictor, which served as a proxy for days post-infection (dpi) or humoral response time in the pig or herd. Regression analysis on individual pig HAIT titres showed that antigen as a predictor, the coefficient for immunogen H1N1pdm09 was at least fourfold higher (P < 0.001) than the three SIVs antigen subtypes, whose much lower coefficients were statistically no different between the three SIVs antigen subtypes. Correspondingly, for herd GMT, immunogen H1N1pdm09 was 28-40-fold higher than the three SIVs antigen subtypes. Excluding the HAIT data of the three SIVs antigen subtypes, regression analysis focusing only on immunogen H1N1pdm09 increased greatly the coefficients of the predictors in the models. Homologous reactions (99.6% H1N1pdm09) have lower HAIT titres while the likelihood of the number of antigens involved in HAIT heterologous reactions in a single pig serum increased with higher HAIT titres of immunogen H1N1pdm09. For predictor 'production', sows and sow herds had higher HAIT titres and GMT compared to fattening pigs and fattening herds respectively. Herds with 'higher proportion of pigs tested positive' also had higher HAIT titre in the pig and herd GMT.

Adverse effects of Influenza A(H1N1)pdm09 virus infection on growth performance of Norwegian pigs - a longitudinal study at a boar testing station.

BACKGROUND: Influenza A(H1N1)pdm09 virus infection in Norwegian pigs was largely subclinical. This study tested the hypothesis that the infection causes negligible impact on pigs' growth performance in terms of feed conversion efficiency, daily feed intake, daily growth, age on reaching 100 kg bodyweight and overall feed intake. A sample of 1955 pigs originating from 43 breeding herds was classified into five infection status groups; seronegative pigs (n = 887); seropositive pigs (n = 874); pigs positive for virus at bodyweight between 33 kg and 60 kg (n = 123); pigs positive for virus at bodyweight between 61 kg and 80 kg (n = 34) and pigs positive for virus at bodyweight between 81 kg and 100 kg (n = 37). Each pig had daily recordings of feed intake and bodyweight from 33 kg to 100 kg. Marginal effects of the virus infection on the outcomes were estimated by multi-level linear regression, which accounted for known fixed effects (breed, birthdate, average daily feed intake and growth phase) and random effects (cluster effects of pig and herd). RESULTS: The seropositive and virus positive pigs had decreased (P value<0.05) growth performance compared to seronegative pigs even though feed intake was not decreased. Reduced feed conversion efficiency led to lower average daily growth, additional feed requirement and longer time needed to reach the 100 kg bodyweight. The effects were more marked (P value<0.03) in pigs infected at a younger age and lasted a longer period. Despite increased feed intake observed, their growth rates were lower and they took more time to reach 100 kg bodyweight compared to the seronegative pigs. CONCLUSION: Our study rejected the null hypothesis that the virus infection had negligible adverse effects on growth performance of Norwegian pigs.

Investigation of an outbreak of mycobacteriosis in pigs.

BACKGROUND: A high proportion of pigs imported to Serbia from a Lithuanian breeding herd reacted positively against avian and/or bovine tuberculin. The pigs were euthanized and lesions characteristic for mycobacterial infection were detected. An investigation of potential mycobacteriosis in the pigs imported to Serbia and the possible source of infection in the Lithuanian herd were therefore initialised. RESULTS: Formalin fixed, paraffin embedded lymph nodes from tuberculin positive animals were examined by real-time PCR for IS1245 and IS6110. IS1245 was detected in 55% and IS6110 in 11% of the samples. Seven of the ten IS6110 positive samples were positive for IS1245. Eleven lymph nodes from 10 pigs and 15 environmental samples were collected from the Lithuanian breeding herd and cultured for mycobacteria. M. avium subsp. hominissuis was detected in all lymph nodes and from eight samples of peat and sawdust. Isolates with identical and related IS1245- and IS1311 RFLP profiles were detected from swine and peat. CONCLUSIONS: This study demonstrated cross reactions between avian and bovine tuberculin in pigs. Real-time PCR indicated infection with M. avium in the Serbian pigs. However, as a small proportion of the lymph nodes were positive for IS6110, infection with bacteria in the M. tuberculosis complex could not be ruled out. Analyses confirmed the presence of M. avium subsp. hominissuis in porcine and environmental samples from the Lithuanian breeding herd. The results indicate peat as a source of M. avium subsp. hominissuis infection in these pigs, and that the pigs imported to Serbia were infected with M. avium subsp. hominissuis.

Successful eradication of Mycoplasma hyopneumoniae from the Norwegian pig population - 10 years later.

BACKGROUND: Mycoplasma hyopneumoniae (Mhyo) is the causative agent of enzootic pneumonia in pigs which adversely affects animal health and welfare, in addition to causing considerable economical losses. This paper presents the implementation of the national Mhyo eradication program in Norway, the subsequent population wide surveillance and documentation on the current freedom from Mhyo in the Norwegian pig population. In 1994, the Board of The Norwegian Pig Health Service decided on conducting a national surveillance and eradication program for Mhyo. The program aimed for population wide freedom from Mhyo, based on serological surveillance. A partial depopulation program was initiated in all Mhyo positive farrow-to-feed and farrow-to-finish herds. Total depopulation was performed in all positive finisher herds. RESULTS: From 1994 to 2009, a total of 138,635 pigs in 3211 herds were serologically tested for the presence of antibodies against Mhyo. Of these, 5538 (4%) individual samples and 398 (12.4%) of the herds were defined as positive. In 2009, the Norwegian pig population was declared free from Mhyo, and has been so since then. From 2009 through 2019, a total of 44,228 individual serum samples have been analyzed for the presence of antibodies against Mhyo and found negative in the National surveillance program. CONCLUSION: Eradication of Mhyo infections has resulted in improved health and welfare of the Norwegian pig population. The success of the strategy is based on numerous factors, such as moderate to low prevalence of the agent, well documented and effective eradication protocols, accurate diagnostic tests, relatively small herds, low herd density in most parts of the country and negligible import of live pigs. In addition, economic benefit due to a premium on pigs marketed from herds free from Mhyo, a well-structured commercial pig population, and finally, the loyalty and significant effort of farmers, abattoir employees and veterinarians were crucial factors. To maintain the infection-free status at national level, a continuous alertness is required in the future to discover possible Mhyo infections and ensure rapid sampling and diagnostics. Any findings of Mhyo positive pig herds in Norway will result in immediate eradication.

Association of myocarditis with high viral load of porcine circovirus type 2 in several tissues in cases of fetal death and high mortality in piglets. A case study.

During a period of 1.5 months, a newly established pig herd experienced a high number of mummifications and stillbirths, a high neonatal mortality rate, and many piglets with congenital tremors or hind leg ataxia. After clinical and histological investigations, the submitted animals were divided into 4 groups: mummified or stillborn (N = 6), live born with myocarditis (N = 5) (average age 22.8 days), live born without myocarditis (N = 14) (average age 20.0 days), and control animals from a different herd (N = 5) (newborn). Statistically significant differences were observed in the mean porcine circovirus 2 (PCV2) load among the 4 groups in the liver (P < 0.0001). The presence of PCV2 antigen within the myocardial lesions was confirmed by immunohistochemistry. A high load of PCV2 DNA was observed in myocardium, liver, and spleen from mummified or stillborn piglets (>1 x 10(7) copies per 500 ng DNA), lower in piglets with myocarditis (>1 x 10(5) copies per 500 ng DNA), and even further lower in pigs without myocarditis (<1 x 10(5) copies per 500 ng DNA), whereas no PCV2 DNA was detected in the control animals. Myocardium, liver, and spleen were well suited for routine testing of fetuses and young piglets by quantitative real-time polymerase chain reaction. Neither porcine parvovirus nor encepaholomyocarditis virus was detected. These results indicate that the PCV2 infection might have been of etiological importance for the fetal deaths and piglet mortality observed in this herd.

Testing of pathogenic Yersinia enterocolitica in pig herds based on the natural dynamic of infection.

This study was performed to evaluate testing methods of pathogenic Yersinia enterocolitica in pigs at different ages. Relevant tools and procedures are crucial if pig herds should be declared free from pathogenic Y. enterocolitica. Historical data based on serology showed that the two farms investigated in this study (herds A and B) were contaminated with Y. enterocolitica O:3 since at least 1995. Laboratory investigations of 60 pigs were sampled one to four times (herd A) and 20 pigs were sampled one to three times (herd B) at different ages were the basis for this report. The following testing procedures could be used to conclude that a herd is free from pathogenic Y. enterocolitica:--serological testing of pigs could be performed as a basis for categorisation for all ages from about 100 days including at slaughter when the pigs are 150-180 days old, --bacteriological examination of faeces could be used as a basis for categorisation at all ages from 85 days until about 135 days, --bacteriological examination of tonsils could be used as a basis for categorisation at all ages from 85 days including at slaughter when the pigs are 150-180 days old. However, due to animal welfare aspects, one should avoid sampling of tonsils. Accordingly, the serological method or bacteriological examination of faeces at relevant ages should be preferred. One aspect related to slaughter hygiene is that in pigs slaughtered at the age of 135 days or more, the tonsils may be a more significant source of human pathogenic Y. enterocolitica than faeces.

Production impact of influenza A(H1N1)pdm09 virus infection on fattening pigs in Norway.

Newly emerged influenza A(H1N1)pdm09 virus infection in Norwegian pigs, although often observed in a subclinical form, can lower the pig's growth performance by reducing feed efficiency in terms of a poorer feed conversion ratio. Infected pigs would consume more feed and require protracted production time to reach market weight. In our observational longitudinal study, growth performance data from 728 control pigs and 193 infected pigs with known viral shedding time points were analyzed using mixed linear regression models to give estimates of the marginal effects of infection. Gaussian curves describing the variability of the estimates at the individual pig level formed the fundamental inputs to our stochastic models. The models were constructed to simulate the summed negative effects of the infection at the batch level of 150 fattening pigs growing from 33 to 100 kg. Other inputs of variability and uncertainty were 1) batch transmission points, 2) pig infection points to reflect the disease transmission dynamics of the virus, and 3) final prevalence of infected pigs in the batch. Monte Carlo random sampling gave 5,000 estimates on the outputs of the marginal effects for each pig. These results were summed up to provide estimates for a batch size of 150 pigs. This figure was adjusted by our final prevalence distribution function, which was also derived from the longitudinal study with 12 cohorts of infected pigs. For a 150-fattening-pig herd randomly selected from the population, the marginal effects of the infection were 1) 835 kg (fifth percentile) to 1,350 kg (95th percentile) increased feed intake and 2) 194 (fifth percentile) to 334 (95th percentile) pig days in excess of expected figures for an uninfected batch. A batch infected during growth phase 3 (81 to 100 kg BW) gave the worst results since the longitudinal study showed that a pig infected during growth phase 3 required more feed and a greater protracted production time compared to younger infected pigs. Sensitivity analysis showed that final prevalence had the greatest impact on the conditional mean and variation of the marginal effects of infections. Batch transmission point was the next most influential factor. Lowering the final prevalence and preventing older fattening pigs from being infected will have the greatest benefit in saving feed cost and reducing delay in getting the pigs to the market.

Mycobacterium avium subsp. hominissuis infection in swine associated with peat used for bedding.

Mycobacterium avium subsp. hominissuis is an environmental bacterium causing opportunistic infections in swine, resulting in economic losses. Additionally, the zoonotic aspect of such infections is of concern. In the southeastern region of Norway in 2009 and 2010, an increase in condemnation of pig carcasses with tuberculous lesions was seen at the meat inspection. The use of peat as bedding in the herds was suspected to be a common factor, and a project examining pigs and environmental samples from the herds was initiated. Lesions detected at meat inspection in pigs originating from 15 herds were sampled. Environmental samples including peat from six of the herds and from three peat production facilities were additionally collected. Samples were analysed by culture and isolates genotyped by MLVA analysis. Mycobacterium avium subsp. hominissuis was detected in 35 out of 46 pigs, in 16 out of 20 samples of peat, and in one sample of sawdust. MLVA analysis demonstrated identical isolates from peat and pigs within the same farms. Polyclonal infection was demonstrated by analysis of multiple isolates from the same pig. To conclude, the increase in condemnation of porcine carcasses at slaughter due to mycobacteriosis seemed to be related to untreated peat used as bedding.

Influenza A(H1N1)pdm09 virus infection in Norwegian swine herds 2009/10: the risk of human to swine transmission.

Influenza A viruses cause respiratory infection in humans and pigs, and some serotypes can be transmitted between these species. The emergence of influenza A(H1N1)pdm09 virus infections in the spring of 2009 quickly led to a worldwide pandemic in humans, with subsequent introduction of the virus to pig populations. Following a widespread infection in the human population in Norway, influenza A(H1N1)pdm09 virus was introduced to the influenza A naïve Norwegian pig population, and within a few months pigs in more than one third of Norwegian swine herds had antibodies against the virus. A cross-sectional study was performed on all swine nucleus and multiplier herds in Norway to analyze risk factors for introduction of infection, and the preventive effects of recommended biosecurity practices. A surveillance program provided information on infection status of the study herds, and a questionnaire was administered to all 118 nucleus and multiplier herds to collect information on herd variables. The surveillance program revealed that pigs in 42% of the herds had antibodies against influenza A(H1N1)pdm09 virus. The incidence of serologically positive pigs was similar in both multiplier herds (41%) and closed nucleus herds (43%). Multivariable logistic regression showed that presence of farm staff with influenza-like illness (ILI) (OR=4.15, CI 1.5-11.4, p=0.005) and herd size (OR=1.01, CI 1-1.02, p=0.009) were risk factors for infection. The rapid and widespread seroconversion for antibodies against influenza A(H1N1)pdm09 virus in the Norwegian pig population can be explained by the emergence of a novel virus that is readily transmitted between people and swine in a largely susceptible population of humans, and an entirely naïve population of pigs.

Pathological Findings and Distribution of Pandemic Influenza A (H1N1) 2009 Virus in Lungs from Naturally Infected Fattening Pigs in Norway.

The Norwegian pig population was considered free from influenza A virus infections until the first case of porcine pandemic influenza A (H1N1) 2009 virus infection in October 2009. Human to pig transmission of virus was suspected. Unusual lung lesions were observed in fattening pigs, with red, lobular, multifocal to coalescing consolidation, most frequently in the cranial, middle, and accessory lobes. The main histopathological findings were epithelial degeneration and necrosis, lymphocyte infiltration in the epithelial lining and lamina propria of small bronchi and bronchioles, and peribronchial and peribronchiolar lymphocyte infiltrations. Infection with pandemic influenza A (H1N1) 2009 virus was confirmed by real-time RT-PCR and immunohistochemical detection of influenza A virus nucleoprotein in the lesions. This investigation shows that natural infection with the pandemic influenza A (H1N1) 2009 virus induces lung lesions similar to lesions described in experimental studies and natural infections with other swine-adapted subtypes of influenza A viruses.

Detection of methicillin-resistant Staphylococcus aureus sequence type 8 in pigs, production environment, and human beings.

Two follow-up studies of a positive methicillin-resistant Staphylococcus aureus (MRSA) finding in the 2008 European Union baseline survey on MRSA in pig herds were performed to gain more knowledge about the epidemiology of the particular MRSA type, a known human type (ST8/t008), among pigs. Two persons on a Norwegian farm in the study were found to be MRSA carriers, and human-to-animal transmission was suspected. In the first follow-up study, all pigs (n  =  346) were sampled by taking nasal swabs. A pooled sample from 5 individual pigs housed together in a single pen, and a dust sample from the equipment in the same room, were positive. Dust samples from a building housing MRSA-negative animals were negative. The MRSA was not detected in the second follow-up, after removing positive animals from the farm and cleaning and disinfecting. A low MRSA occurrence among the animals was found, suggesting that MRSA ST8/t008 may be less able to colonize and persist in pig holdings compared with more host-adapted S. aureus strains.

Clinical Impact of Infection with Pandemic Influenza (H1N1) 2009 Virus in Naïve Nucleus and Multiplier Pig Herds in Norway.

The Norwegian pig population has been free from influenza viruses until 2009. The pandemic influenza outbreak during the autumn 2009 provided an opportunity to study the clinical impact of this infection in an entirely naïve pig population. This paper describes the results of a case-control study on the clinical impact of pandemic influenza (H1N1) 2009 infection in the nucleus and multiplier herds in Norway. The infection spread readily and led to seroconversion of 42% of the Norwegian nucleus and multiplier herds within a year. Positive and negative herds were identified based on surveillance data from the Norwegian Veterinary Institute. Telephone interviews were conducted with pig herd owners or managers between November 2010 and January 2011. Pigs with clinical signs were reported from 40% of the case herds with varying morbidity and duration of respiratory disease and reduced reproductive performance. Clinical signs were reported in all age groups.

Dynamics of serum antibodies to and load of porcine circovirus type 2 (PCV2) in pigs in three finishing herds, affected or not by postweaning multisystemic wasting syndrome.

BACKGROUND: Despite that PMWS commonly affects pigs aged eight to sixteen weeks; most studies of PMWS have been conducted during the period before transfer to finishing herds. This study focused on PCV2 load and antibody dynamics in finishing herds with different PMWS status. METHODS: Sequentially collected blood samples from 40 pigs in each of two Swedish (A and B) and one Norwegian (C) finishing herds were analysed for serum PCV2-load and -antibodies and saliva cortisol. The two Swedish herds differed in PMWS status, despite receiving animals from the same sow pool (multi-site production). However, the PMWS-deemed herd (A) had previously also received pigs from the spot market. RESULTS: The initial serum PCV2 load was similar in the two Swedish herds. In herd A, it peaked after two weeks in the finishing herd and a high number of the pigs had serum PCV2 levels above 107 per ml. The antibody titres increased continually with exception for the pigs that developed PMWS, that had initially low and then declining antibody levels. Pigs in the healthy herd B also expressed high titres of antibodies to PCV2 on arrival but remained at that level throughout the study whereas the viral load steadily decreased. No PCV2 antibodies and only low amounts of PCV2 DNA were detected in serum collected during the first five weeks in the PMWS-free herd C. Thereafter a peak in serum PCV2 load accompanied by an antibody response was recorded. PCV2 from the two Swedish herds grouped into genotype PCV2b whereas the Norwegian isolate grouped into PCV2a. Cortisol levels were lower in herd C than in herds A and B. CONCLUSIONS: The most obvious difference between the Swedish finishing herds and the Norwegian herd was the time of infection with PCV2 in relation to the time of allocation, as well as the genotype of PCV2. Clinical PMWS was preceded by low levels of serum antibodies and a high load of PCV2 but did not develop in all such animals. It is notable that herd A became affected by PMWS after errors in management routine, emphasising the importance of proper hygiene and general disease-preventing measures.

Pig herds free from human pathogenic Yersinia enterocolitica.

Pork products are a substantial source of human yersiniosis, a foodborne disease caused by Yersinia enterocolitica. Thus, the ability to eliminate this agent from pig herds would be an important step in producing human pathogen-free pork. Pig herds free from Y. enterocolitica O:3/biovar 4 have been established and maintained. According to serologic and cultural testing results, 15 of 16 specific pathogen-free herds were free from Y. enterocolitica O:3/biovar 4; this closed breeding pyramid has remained free from this organism since 1996. Pig herds free from human pathogenic Y. enterocolitica suggest that human pathogen-free herds could be attained to provide pork free from zoonotic agents.