The expression of sexually selected traits often varies with populations’ breeding cycles in many animals. The elucidation of mechanisms underlying the expression of such traits is a research topic in evolutionary biology; however, the genetic basis of the seasonal development of their expression remains unknown. Male Leptobrachium boringii develop keratinized nuptial spines on their upper jaw during the breeding season that fall off when the breeding season ends. To illuminate the genetic basis for the expression of this trait and its seasonal development, we assessed the de novo transcriptome for L. boringii using brain, testis and upper jaw skin and compared gene expression profiles of these tissues between two critical periods of the spine growth cycle. We identified 94,900 unigenes in our transcriptome. Among them, 2,131 genes were differentially expressed between the breeding period when the spines developed and the post-breeding period when the spines were sloughed. An increased number of differentially expressed genes (DEGs) were identified in the upper jaw skin compared with the testis and brain. In the upper jaw skin, DEGs were mainly enriched in cytosolic part, peptidase inhibitor activity and peptidase regulator activity based on GO enrichment analysis and in glycolysis/gluconeogenesis, ribosome biogenesis in eukaryotes and retinol metabolism based on KEGG enrichment analysis. In the other two tissues, DEGs were primarily involved in the cell cycle, DNA replication and melatonin production. Specifically, insulin/insulin-like growth factor and sex steroid hormone-related DEGs were identified in the upper jaw skin. The expression variation of IGF2 and estrogen-related genes may be the main factors regulating the seasonal development of the spines. Our study provides a list of potential genes involved in the regulation of seasonal development of nuptial spines in L. boringii. This is the first transcriptome survey of seasonally developed sexually selected traits for non-model amphibian species, and candidate genes provided here may provide valuable information for further studies of L. boringii.

Rutin, a natural flavonol glycoside, elicits its diverse health-promoting effects from the bioactivities of quercetin, its aglycone. While widely distributed in the vegetables and fruits of human diet, rutin is either absent or inadequate in common animal feed ingredients. Rutin has been supplemented to dairy cows for performance enhancement, but its metabolic fate in vivo has not been determined. In this study, plasma, urine, and rumen fluid samples were collected before and after the intraruminal dosing of 100 mg/kg rutin to 4 Holsteins, and then characterized by both targeted and untargeted liquid chromatography-mass spectrometry (LC-MS)-based metabolomic analysis. In plasma and urine, 4-methylcatechol sulfate was identified as the most abundant metabolite of rutin, instead of quercetin and its flavonol metabolites, and its concentration was inversely correlated with the concentration of p-cresol sulfate. In rumen fluid, the formation of 3,4-dihydroxyphenylacetic acid (DHPAA) and 4-methylcatechol after rapid degradation of rutin and quercetin concurred with the decrease of p-cresol and the increase of its precursor, 4-hydroxyphenylacetic acid. Overall, the formation of 4-methylcatechol, a bioactive microbial metabolite, as the dominant bioavailable metabolite of rutin and quercetin, could contribute to their beneficial bioactivities in dairy cows, while the decrease of p-cresol, a microbial metabolite with negative biological and sensory properties, from the competitive inhibition between microbial metabolism of rutin and tyrosine, has the potential to reduce environmental impact of dairy operations and improve the health of dairy cattle.

Lawsonia intracellularis, an obligate intracellular bacterium, is an important enteric pathogen in pig herds and horse farms worldwide. The hallmark feature of L. intracellularis infection is the proliferation of epithelial cells in intestinal crypts. A major limitation to the study of L. intracellularis infection is the lack of an in vitro model that reproduces the changes observed in proliferative enteropathy. Here we investigated the suitability of mouse enteroids as a model to study L. intracellularis infection. Mouse enteroids were microinjected with L. intracellularis, filter-sterilized L. intracellularis culture supernatant, or sterile cell culture media (DMEM). L. intracellularis antigen was detected in mouse enteroids by immunohistochemistry and was located mostly in the basal region of the epithelium. There was no differential growth of enteroids among treatment groups, and cellular proliferation was not increased in L. intracellularis-infected enteroids in relation to non-infected enteroids based on immunofluorescence staining. L. intracellularis infection did not induce changes in gene expression of Ki-67 (proliferation marker), Sox9 (marker for transit amplifying cells) and Muc2 (marker for goblet cells). These results indicate that although L. intracellularis antigen is detectable in mouse enteroids, indicating susceptibility to infection, mouse enteroids fail to replicate the cellular proliferation and gene expression changes observed in proliferative enteropathy. Nevertheless, we have successfully demonstrated that mouse enteroids can be used to model days-long intracellular pathogen infection, serving as potential models for the study of other pathogens of interest in veterinary medicine.

Animals receiving Zinc (Zn) dietary supplementation with organic sources respond better to stress than inorganic Zn sources supplementation. The study aimed to identify the effect of different Zn sources on intestinal epithelial gene expression. In total, 45 pigs (9 per treatment) (77.5 ± 2.5 kg weight) were fed for 32 days, a corn-soybean meal diet without supplemented Zn (ZnR) or supplemented with 50 and 100 ppm of inorganic ZnCl₂ (Zn50 and Zn100), and amino acid-bound organic Zn sources (LQ50 and LQ100). Gene expression changes form RNA-seq in ileum tissues of ZnR revealed changes associated with Zn insufficiency. Comparing organic with inorganic Zn sources by one-way ANOVA, pro-inflammatory cytokine interleukin 18 (IL18) was downregulated (p = 0.03) and Toll-like receptor 2 (TLR2) upregulated (p = 0.02). To determine the role of epithelial cells in response to dietary Zn, swine intestinal organoids (enteroids) were exposed to Zn restriction, ZnCl₂ or LQ-Zn. In enteroids, ZIP4 expression decreased with added Zn compared with Zn-restriction (p = 0.006) but Zn sources did not affect (p > 0.05) IL18 or TLR2 expression. These results suggest that organic Zn may stimulate TLR2 signaling possibly affecting immune response, while decreasing the proinflammatory cytokine IL18 expression in non-epithelial cells of intestinal mucosa.

Feeding thermally oxidized lipids to pigs has been shown to compromise growth and health, reduce energy digestibility, and disrupt lipid metabolism. However, the effects of feeding oxidized lipids on amino acid metabolism in pigs have not been well defined even though amino acids are indispensable for the subsistence of energy metabolism, protein synthesis, the antioxidant system, and many other functions essential for pig growth and health. In this study, oxidized corn oil (OCO)-elicited changes in amino acid homeostasis of nursery pigs were examined by metabolomics-based biochemical analysis. The results showed that serum and hepatic free amino acids and metabolites, including tryptophan, threonine, alanine, glutamate, and glutathione, as well as associated metabolic pathways, were selectively altered by feeding OCO, and more importantly, many of these metabolic events possess protective functions. Specifically, OCO activated tryptophan-nicotinamide adenosine dinucleotide (NAD⁺) synthesis by the transcriptional upregulation of the kynurenine pathway in tryptophan catabolism and promoted adenine nucleotide biosynthesis. Feeding OCO induced oxidative stress, causing decreases in glutathione (GSH)/oxidized glutathione (GSSG) ratio, carnosine, and ascorbic acid in the liver but simultaneously promoted antioxidant responses as shown by the increases in hepatic GSH and GSSG as well as the transcriptional upregulation of GSH metabolism-related enzymes. Moreover, OCO reduced the catabolism of threonine to α-ketobutyrate in the liver by inhibiting the threonine dehydratase (TDH) route. Overall, these protective metabolic events indicate that below a certain threshold of OCO consumption, nursery pigs are capable of overcoming the oxidative stress and metabolic challenges posed by the consumption of oxidized lipids by adjusting antioxidant, nutrient, and energy metabolism, partially through the transcriptional regulation of amino acid metabolism.

Background: Studies of Positively Selected Genes (PSGs) in microorganisms and mammals have provided insights into the dynamics of genome evolution and the genetic basis of differences between species by using whole genome-wide scans. Systematic investigations and comparisons of PSGs in plants, however, are still limited.Objective: A systematic comparison of PSGs between the genomes of two cotton species, Gossypium arboreum (G. arboreum) and G. raimondii, will give the key answer for revealing molecular evolutionary differences in plants.Methods: Genome sequences of G. arboreum and G. raimondii were compared, including Whole Genome Duplication (WGD) events and genomic features such as gene number, gene length, codon bias index, evolutionary rate, number of expressed genes, and retention of duplicated copies.Results: Unlike the PSGs in G. raimondii, G. arboreum comprised more PSGs, smaller gene size and fewer expressed gene. In addition, the PSGs evolved at a higher rate of synonymous substitutions, but were subjected to lower selection pressure. The PSGs in G. arboreum were also retained with a lower number of duplicate gene copies than G. raimondii after a single WGD event involving Gossypium.Conclusion: These data indicate that PSGs in G. arboreum and G. raimondii differ not only in Ka/Ks, but also in their evolutionary, structural, and expression properties, indicating that divergence of G. arboreum and G. raimondii was associated with differences in PSGs in terms of evolutionary rates, gene length, expression patterns, and WGD retention in Gossypium.