Gene expression in hypothalamus, liver, and adipose tissues and food intake response to melanocortin-4 receptor agonist in pigs expressing melanocortin-4 receptor mutations

Transcriptional profiling was used to identify genes and pathways that responded to intracerebroventricular injection of melanocortin-4 receptor (MC4R) agonist [Nle4, D-Phe7]-α-melanocyte stimulating hormone (NDP-MSH) in pigs homozygous for the missense mutation in the MC4R, D298 allele (n = 12), N298 allele (n = 12), or heterozygous (n = 12). Food intake (FI) was measured at 12 and 24 h after treatment. All pigs were killed at 24 h after treatment, and hypothalamus, liver, and back-fat tissue was collected. NDP-MSH suppressed (P \u3c 0.004) FI at 12 and 24 h in all animals after treatment. In response to NDP-MSH, 278 genes in hypothalamus (q ≤ 0.07, P ≤ 0.001), 249 genes in liver (q ≤ 0.07, P ≤ 0.001), and 5,066 genes in fat (q ≤ 0.07, P ≤ 0.015) were differentially expressed. Pathway analysis of NDP-MSH-induced differentially expressed genes indicated that genes involved in cell communication, nucleotide metabolism, and signal transduction were prominently downregulated in the hypothalamus. In both liver and adipose tissue, energy-intensive biosynthetic and catabolic processes were downregulated in response to NDP-MSH. This included genes encoding for biosynthetic pathways such as steroid and lipid biosynthesis, fatty acid synthesis, and amino acid synthesis. Genes involved in direct energy-generating processes, such as oxidative phosphorylation, electron transport, and ATP synthesis, were upregulated, whereas TCA-associated genes were prominently downregulated in NDP-MSH-treated pigs. Our data also indicate a metabolic switch toward energy conservation since genes involved in energy-intensive biosynthetic and catabolic processes were downregulated in NDP-MSH-treated pigs

GENOMICS SYMPOSIUM: Using genomic approaches to uncover sources of variation in age at puberty and reproductive longevity in sows

Genetic variants associated with traits such as age at puberty and litter size could provide insight into the underlying genetic sources of variation impacting sow reproductive longevity and productivity. Genomewide characterization and gene expression profiling were used using gilts from the University of Nebraska–Lincoln swine resource population (n = 1,644) to identify genetic variants associated with age at puberty and litter size traits. From all reproductive traits studied, the largest fraction of phenotypic variation explained by the Porcine SNP60 BeadArray was for age at puberty (27.3%). In an evaluation data set, the predictive ability of all SNP from highranked 1-Mb windows (1 to 50%), based on genetic variance explained in training, was greater (12.3 to 36.8%) compared with the most informative SNP from these windows (6.5 to 23.7%). In the integrated data set (n = 1,644), the top 1% of the 1-Mb windows explained 6.7% of the genetic variation of age at puberty. One of the high-ranked windows detected (SSC2, 12–12.9 Mb) showed pleiotropic features, affecting both age at puberty and litter size traits. The RNA sequencing of the hypothalami arcuate nucleus uncovered 17 differentially expressed genes (adjusted P \u3c 0.05) between gilts that became pubertal early (180 d of age). Twelve of the differentially expressed genes are upregulated in the late pubertal gilts. One of these genes is involved in energy homeostasis (FFAR2), a function in which the arcuate nucleus plays an important contribution, linking nutrition with reproductive development. Energy restriction during the gilt development period delayed age at puberty by 7 d but increased the probability of a sow to produce up to 3 parities (P \u3c 0.05). Identification of pleotropic functional polymorphisms may improve accuracy of genomic prediction while facilitating a reduction in sow replacement rates and addressing welfare concerns

Litter-of-origin trait effects on gilt development

The preweaning litter environment of gilts can affect subsequent development. In a recent experiment designed to test the effects of diet on gilt development, litter-of-origin traits including individual birth weights, immunocrits (a measure of colostrum intake), sow parity, number weaned, and individual weaning weights were collected for approximately 1,200 gilts that were progeny of approximately 300 sows. Subsequently, BW, LM area, and backfat were measured at 100 d of age and at 28-d intervals until slaughter (260 d of age). From 160 d of age to slaughter, gilts were observed daily for estrus. At slaughter, the reproductive tract and 1 mammary gland were recovered. The reproductive tract was classified as cyclic or prepubertal; the number of corpora lutea was counted. Uterine horn lengths and ovarian dimensions were measured. Uterus and ovary samples from every 10th gilt were prepared for histological evaluation of uterine gland development and follicle counts, respectively. Mammary gland tissue protein and fat were assayed. Day of the estrous cycle at slaughter was calculated using the first day of the most recent standing estrus (d 0) recorded previous to slaughter. Each gilt development trait was analyzed for association with each litter-of-origin trait, after adjusting for dietary treatment effects. Uterine length, ovarian dimensions, mammary gland protein and fat, and uterine gland development were also adjusted for day of the estrous cycle at slaughter. All litter-of-origin traits were associated (P \u3c 0.05) with growth traits. Top-down (backward elimination) multiple regression analysis indicated that BW and LM accretion in gilts was positively associated with immunocrit (P \u3c 0.01), birth weight (P \u3c 0.01), preweaning growth rate (P \u3c 0.01), and parity (P \u3c 0.01). Backfat accretion was positively associated with preweaning growth rate (P \u3c 0.01), number weaned (P\u3c 0.05), and parity (P \u3c 0.05). Age at puberty was associated with birth weight (positive; P \u3c 0.01) and preweaning growth rate (negative; P \u3c 0.01). Total uterine length was positively associated with only birth weights (P \u3c 0.05). Mammary gland protein was negatively associated with preweaning growth (P\u3c 0.01). Mammary gland fat was positively associated with birth weight and number of piglets weaned (P \u3e 0.05). These results indicate that colostrum consumption, birth weights, preweaning growth rate, number weaned, and parity are associated with gilt development traits during later life

Age at puberty, ovulation rate, and uterine length of developing gilts fed two lysine and three metabolizable energy concentrations from 100 to 260 d of age

The objective of this study was to determine the effect of ad libitum feeding diets differing in standard ileal digestible (SID) lysine and ME concentrations that bracket those fed to developing gilts in U.S. commercial settings. Average SID lysine and ME concentrations in diets currently fed to developing gilts were obtained from a poll of the U.S. commercial swine industry. Crossbred Large White × Landrace gilts (n = 1,221), housed in groups, were randomly allotted to 6 corn-soybean diets in a 2 × 3 factorial arrangement formulated to provided 2 SID lysine and 3 ME concentrations. Gilts received grower diets formulated to provide 1.02% (control = survey average) or 0.86% (control minus 15%) SID lysine and 2.94, 3.25, or 3.57 (survey average ME ± 10%) Mcal of ME/kg from 100 d of age until approximately 90 kg BW. Then, gilts were fed finisher diet containing 0.85% (control = survey average) or 0.73% (control minus 15%) SID lysine and 2.94, 3.26, or 3.59 (control ± 10%) Mcal of ME/kg until 260 d of age. Gilts were weighed, and backfat thickness and loin muscle area were recorded at the beginning of the trial and then every 28 d. Starting at 160 d of age, gilts were exposed daily to vasectomized boars and observed for behavioral estrus. At approximately 260 d of age, gilts were slaughtered and their reproductive tract was collected. Each reproductive tract was examined to determine whether the gilt was cyclic, the stage of estrus cycle, ovulation rate, and uterine length. Data were evaluated for normality and analyzed using mixed model methods. Average age at puberty was 193 d of age with a range from 160 to 265 d. When all gilts on trial at 160 d of age were included in the analysis, 91.0% reached puberty as determine by observation of standing estrus. Differences between dietary treatments on age at puberty or measurements of the reproductive tract were not detected. Growth rates to 160 d were not limiting for attainment of puberty in response to daily boar stimulation from 160 d

The roles of age at puberty and energy restriction |in sow reproductive longevity: a genomic perspective

Approximately 50% of sows are culled annually with more than one-third due to poor fertility. Our research demonstrated that age at puberty is an early pre-breeding indicator of reproductive longevity. Age at puberty can be measured early in life, has a moderate heritability, and is negatively correlated with lifetime number of parities. Detection of age at puberty is tedious and time consuming and is therefore not collected by the industry, which limits genetic progress. Genomic prediction is a viable approach to preselect gilts that will express puberty early and have superior reproductive longevity. The hypothesis that genetic variants explaining differences in age at puberty also explain differences in sow reproductive longevity was tested. Phenotypes, genotypes, and tissues from the UNL resource population (n \u3e 1700) were used in genome-wide association analyses, genome, and RNA sequencing to uncover functional polymorphisms that could explain variation in puberty and reproductive longevity. A BeadArray including 56,424 SNP explained 25.2% of the phenotypic variation in age at puberty in a training set (n = 820). Evaluation of major windows and SNPs of subsequent batches of similar genetics (n = 412) showed that if all SNPs located in the major 1-Mb windows were tested, they explained a substantial amount of phenotypic variation (12.3 to 36.8%). Due to differences in linkage disequilibrium status, the most informative SNP from these windows explained a lower proportion of the variation (6.5 to 23.7%). To improve genomic predictive ability, the limited capability of BeadArray was enhanced by potential functional variants uncovered by genome sequencing of selected sires (n = 20; \u3e20X). There were 11.2 mil. SNPs and 2.9 mil. indels discovered across sires and reference genomes. The role of gene expression differences in explaining phenotypic variation in age at puberty was investigated by RNA sequencing of the hypothalamic arcuate nucleus (ARC) in gilts (n = 37) with different pubertal statuses. Seventy genes, including genes involved in reproductive processes, were differentially expressed between gilts with early and late puberty status (Padj \u3c 0.1). Dietary restriction of energy 3 mo before breeding delayed puberty by 7 d but improved the potential of a sow producing up to three parities (P \u3c 0.05). Energy restriction was associated with differential expression in 42 genes in the ARC, including genes involved in energy metabolism. This integrated genomic information will be evaluated in commercial populations to improve the reproductive potential of sows through genomic selection. This project is supported by AFRI Competitive grant no. 2013-68004-20370 from the USDA-NIFA. USDA is an equal opportunity provider and employer

Dietary lysine-to-energy ratios for managing growth and pubertal development in replacement gilts

Objective: Our objective was to determine growth rates, body composition, and pubertal development of replacement gilts fed diets with different ratios of standardized ileal digestible (SID) lysine to ME. Materials and Methods: Diets with low, medium, and high ratios of SID lysine to ME (grower: 2.3, 2.6, and 2.8; and, finisher; 1.7, 1.9, and 2.1 g/Mcal) were fed from 100 to 200 d of age, after which gilts were moved from the gilt development unit to sow farms. Boar exposure and estrus detection began at 160 d of age and continued until first detected estrus. Estimates of BW and body composition were determined at 100, 142, 160, and 200 d of age and at puberty. Results and Discussion: Body weights and growth rates were reduced (P \u3c 0.05) as dietary SID lysine–to– ME ratio decreased. Greater SID lysine–to–ME ratios increased the number of gilts that exhibited estrus upon boar exposure, increased the number of gilts with a spontaneous first estrus, reduced the number of gilts requiring P.G.600 (Merck Animal Health, Kenilworth, NJ), and decreased age at first estrus (P \u3c 0.05). Slower growing gilts that weighed less with less backfat were more likely to require P.G.600 to induce puberty (P \u3c 0.05). Implications and Applications: Reducing SID lysine– to–ME ratios in gilt diets can increase the number of gilts within optimal BW range at first estrus, but overall pubertal development is delayed if ratios are reduced below 2.8 and 2.1 g of SID lysine to megacalorie of ME in grower and finisher diets, respectively

Energy balance affects pulsatile secretion of luteinizing hormone from the adenohypophesis and expression of neurokinin B in the hypothalamus of ovariectomized gilts

The pubertal transition of gonadotropin secretion in pigs is metabolically gated. Kisspeptin (KISS1) and neurokinin B (NKB) are coexpressed in neurons within the arcuate nucleus of the hypothalamus (ARC) and are thought to play an important role in the integration of nutrition and metabolic state with the reproductive neuroendocrine axis. The hypothesis that circulating concentrations of luteinizing hormone (LH) and expression of KISS1 and tachykinin 3(TAC3, encodes NKB) in the ARC of female pigs are reduced with negative energy balance was tested using ovariectomized, prepubertal gilts fed to either gain or lose body weight. Restricted feeding of ovariectomized gilts caused a rapid and sustained metabolic response characterized by reduced concentrations of plasma urea nitrogen, insulin, leptin, and insulin-like growth factor-1 and elevated concentrations of free fatty acids. The secretory pattern of LH shifted from one of low amplitude to one of high amplitude, which caused overall circulating concentrations of LH to be greater in restricted gilts. Nutrient-restricted gilts had greater expression of follicle-stimulating hormone and gonadotropinreleasing hormone receptor, but not LH in the anterior pituitary gland. Expression of KISS1 in the ARC was not affected by dietary treatment, but expression of TAC3 was greater in restricted gilts. These data are consistent with the idea that hypothalamic expression of KISS1 is correlated with the number of LH pulse in pig, and further indicate that amplitude of LH pulses may be regulated by NKB in the gilt

Treatment of mastitis during lactation

Treatment of mastitis should be based on bacteriological diagnosis and take national and international guidelines on prudent use of antimicrobials into account. In acute mastitis, where bacteriological diagnosis is not available, treatment should be initiated based on herd data and personal experience. Rapid bacteriological diagnosis would facilitate the proper selection of the antimicrobial. Treating subclinical mastitis with antimicrobials during lactation is seldom economical, because of high treatment costs and generally poor efficacy. All mastitis treatment should be evidence-based, i.e., the efficacy of each product and treatment length should be demonstrated by scientific studies. Use of on-farm written protocols for mastitis treatment promotes a judicious use of antimicrobials and reduces the use of antimicrobials