A first-generation porcine oligonucleotide set, representing 13,297 cDNAs and ESTs, has been designed by Qiagen-Operon for transcriptional profiling. To validate this set, microarrays containing each 70-mer oligonucleotide, referred to as the Qiagen-NRSP8 array, were hybridized with targets from porcine adult liver, lung, muscle, or small intestine. Transcriptome analyses showed that 11,328 of the oligonucleotides demonstrated expression in at least one tissue. Statistical analyses revealed that 1810 genes showed differential expression among tissues (Bonferroni adjusted p < 0.05). Biological pathways identified by DAVID/EASE analysis using a list of 423 tissue-selective genes matched archetypal pathways in the corresponding human or mouse tissue. Real-time quantitative PCR confirmed expression patterns for 9 of 11 genes tested. Our results demonstrate that this first-generation porcine oligonucleotide array is informative and the specificity is high. This is essential validation for investigators using the Qiagen-NRSP8 array for porcine functional genomics and for using the pig in modeling important physiological problems.

Infection with porcine reproductive and respiratory syndrome virus (PRRSV) results in a weak antiviral immune response that leads to a persistent infection in a subset of pigs. We investigated the intensity and timing of the early cytokine responses to PRRSV infection to determine their utility as a predictor of persistence. As part of the ''Big Pig'' project, we evaluated cytokine gene expression in lymphoid tissues collected from pigs for up 202 days post-infection (dpi); serum samples were collected biweekly. Cytokine mRNA levels were compared between pigs that cleared the viral infection from serum and tissues (non-persistent [NP] pigs) to those of persistent (P) pigs, that had viral RNA in their serum for up to 126 dpi. The gene expression studies in the tracheobronchial lymph nodes (TBLN) of all the pigs showed upregulation of interferon-g (IFN-g)-associated T-helper 1 (Th-1) markers from 14-84 dpi, and of T-regulatory interleukin-10 (IL-10), but no upregulation of innate markers (IFN-A, IL-1B, and IL-8). At later time points (>112 dpi) these genes were no longer differentially expressed and thus were uninformative for persistence studies. Statistical analyses of serum cytokine levels indicated that innate cytokine (IL-1b and IL-8) levels were upregulated early after infection. Interestingly, serum IL-8 levels in NP pigs were significantly higher than in P pigs at 14 dpi. When analyzed together, variations in all three of the serum cytokines tested (IL-8, IL-1b, and IFN-g) was significantly correlated with virus level, accounting for *84% of the variations observed. These results indicate that while each cytokine individually has minor effects on the length of virus replication, the combination of cytokine activities should be considered when understanding the role of immunity in persistence.

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Halyomorpha halys (Stål) (Heteroptera: Pentatomidae), the brown marmorated stink bug, is an invasive agricultural and nuisance pest rapidly expanding its incidence in North America. This voracious pest poses a significant threat to rural and urban agriculture, especially to specialty crops such as apples, grapes and ornamentals, as well as staple crops including soybean and corn. The object of this study was to generate transcript sequence resources for H. halys. RNA-seq libraries derived from distinct developmental stages and sexes were sequenced and assembled into 248,569 putatively unique transcripts (PUTs). PUTs were segmented into three disjoint tiers of varying reliability, with 4,794 classified as gold tier (highest quality), 16,878 as silver, and 14,357 as bronze. The gold-tier PUTs associated with 2,580 distinct non-redundant protein sequences from the NCBI NR database—1,785 of these (69%) mapped to annotated UniProtKB database proteins, from which 1,273 unique Pfam families and 459 unique Molecular Function GO terms were encountered. Of the silver tier's 6,527 PUTs associated with unique proteins, 4,193 mapped to UniProtKB (64%), from which 1,941 and 640 unique Pfam and Molecular Function GO terms were extracted. H. halys PUTs related to important life processes like immunity, endocrinology, reproduction, development, behavior, neurotransmission, neurotoxicity, olfaction, and small RNA pathways were validated through quantitative Real-Time PCR (qRT-PCR) for differential expression during distinct life stages (eggs, 2nd instar nymphs, 4th instar nymphs, female adults, male adults). PUTs similar to hypothetical proteins identified in symbiont microbes, including Pantoea and Nosema species, were more abundantly expressed in adults versus nymphs. These comprehensive H. halys transcriptomic resources can be utilized to aid development of novel control methodologies to disrupt life processes; to conduct reverse genetic screens to determine host gene function; and to design environmentally unobtrusive means to control host populations or target specific H. halys life stages, such as molecular biopesticides.