Background: A major determinant of influenza infection is the presence of virus receptors on susceptible host cells to which the viral haemagglutinin is able to bind. Avian viruses preferentially bind to sialic acid α2,3-galactose (SAα2,3-Gal) linked receptors, whereas human strains bind to sialic acid α2,6-galactose (SAα2,6-Gal) linked receptors. To date, there has been no detailed account published on the distribution of SA receptors in the pig, a model host that is susceptible to avian and human influenza subtypes, thus with potential for virus reassortment. We examined the relative expression and spatial distribution of SAα2,3-GalG(1-3)GalNAc and SAα2,6-Gal receptors in the major organs from normal post-weaned pigs by binding with lectins Maackia amurensis agglutinins (MAA II) and Sambucus nigra agglutinin (SNA) respectively. Results: Both SAα2,3-Gal and SAα2,6-Gal receptors were extensively detected in the major porcine organs examined (trachea, lung, liver, kidney, spleen, heart, skeletal muscle, cerebrum, small intestine and colon). Furthermore, distribution of both SA receptors in the pig respiratory tract closely resembled the published data of the human tract. Similar expression patterns of SA receptors between pig and human in other major organs were found, with exception of the intestinal tract. Unlike the limited reports on the scarcity of influenza receptors in human intestines, we found increasing presence of SAα2,3-Gal and SAα2,6-Gal receptors from duodenum to colon in the pig. Conclusions: The extensive presence of SAα2,3-Gal and SAα2,6-Gal receptors in the major organs examined suggests that each major organ may be permissive to influenza virus entry or infection. The high similarity of SA expression patterns between pig and human, in particular in the respiratory tract, suggests that pigs are not more likely to be potential hosts for virus reassortment than humans. Our finding of relative abundance of SA receptors in the pig intestines highlights a need for clarification on the presence of SA receptors in the human intestinal tract.

One requisite of quantitative reverse transcription PCR (qRT-PCR) is to normalise the data with an internal reference gene that is invariant regardless of treatment, such as virus infection. Several studies have found variability in the expression of commonly used housekeeping genes, such as beta-actin (ACTB) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), under different experimental settings. However, ACTB and GAPDH remain widely used in the studies of host gene response to virus infections, including influenza viruses. To date no detailed study has been described that compares the suitability of commonly used housekeeping genes in influenza virus infections. The present study evaluated several commonly used housekeeping genes [ACTB, GAPDH, 18S ribosomal RNA (18S rRNA), ATP synthase, H+ transporting, mitochondrial F1 complex, beta polypeptide (ATP5B) and ATP synthase, H+ transporting, mitochondrial Fo complex, subunit C1 (subunit 9) (ATP5G1)] to identify the most stably expressed gene in human, pig, chicken and duck cells infected with a range of influenza A virus subtypes.

Aquatic birds are the natural reservoir for most subtypes of influenza A, and a source of novel viruses with the potential to cause human pandemics, fatal zoonotic disease or devastating epizootics in poultry. It is well recognised that waterfowl typically show few clinical signs following influenza A infection, in contrast, terrestrial poultry such as chickens may develop severe disease with rapid death following infection with highly pathogenic avian influenza. This study examined the cellular response to influenza infection in primary cells derived from resistant (duck) and susceptible (chicken) avian hosts. Paradoxically, we observed that duck cells underwent rapid cell death following infection with low pathogenic avian H2N3, classical swine H1N1 and ‘classical’ highly pathogenic H5N1 viruses. Dying cells showed morphological features of apoptosis, increased DNA fragmentation and activation of caspase 3/7. Following infection of chicken cells, cell death occurred less rapidly, accompanied by reduced DNA fragmentation and caspase activation. Duck cells produced similar levels of viral RNA but less infectious virus, in comparison with chicken cells. Such rapid cell death was not observed in duck cells infected with a contemporary Eurasian lineage H5N1 fatal to ducks. The induction of rapid death in duck cells may be part of a mechanism of host resistance to influenza A, with the loss of this response leading to increased susceptibility to emergent strains of H5N1. These studies provide novel insights that should help resolve the long‐standing enigma of host–pathogen relationships for highly pathogenic and zoonotic avian influenza.

There is now an increasing debate about the viability of using temperate-grown legumes in pig diets as a potential replacement for imported soya bean meal (SBM) and this is due to food security, sustainability and environmental concerns. Two trials were designed to examine nitrogen (N) retention, growth performance and carcass quality of grower and finisher pigs when fed nutritionally balanced SBM-free diets formulated to contain peas or faba beans at 300 g/kg, compared to an SBM-containing, pulse-free control diet. Trial 1 evaluated N digestibility/retention in four iso-energetic diets, comparing the SBM control with one diet formulated with peas and two with faba bean cultivars; a tannin-containing and a tannin-free variety. This trial employed a four by four Latin Square design with four male pigs housed in metabolism crates, fed twice daily at 0.9 of assumed ad libitum intake over four time periods during grower (30–55 kg) and finisher (55–95 kg) phases. Quantitative faecal and urine collection allowed determination of N coefficient of total tract apparent digestibility, coefficient of apparent metabolisability, and N balance. Results revealed that dietary treatment did not affect these N parameters (P > 0.05) during either the grower or finisher phase. Trial 2 evaluated growth performance (feed intake, daily live weight gain and feed conversion ratio) and carcass quality parameters. Five diets (based on SBM, peas and one of three faba bean cultivars) balanced for standard ileal digestible amino acids and net energy were each fed to eight replicates of individually housed entire male pigs over the same growth phases as Trial 1. The inclusion of three faba bean varieties allowed comparison of animal responses between tannin/tannin-free and spring vs. winter bean cultivars. At ∼95 kg, pigs were slaughtered and a comprehensive range of carcass measurements undertaken. Samples of shoulder backfat were also taken at slaughter to determine skatole and indole concentrations. As with N balance, feeding treatment did not affect performance data. Carcass parameters revealed pigs fed with the pea-based diet had a greater dressing percentage than those animals on faba bean-based diets. Pigs fed with the SBM or pea-based diets also had greater lean meat percentages than those on faba-bean diets. Mean skatole concentrations for all pigs were below the accepted maximum threshold level of 0.2 μg/g. In conclusion, it is suggested that peas and faba beans can be successfully fed in balanced pig diets throughout the grower/finisher periods as alternatives to SBM