W. T. Oliver

Adipose and Muscle Tissue Gene Expression of Two Genes (NCAPG and LCORL) Located in a Chromosomal Region Associated with Cattle Feed Intake and Gain

A region on bovine chromosome 6 has been implicated in cattle birth weight, growth, and length. Non-SMC conodensin I complex subunit G (NCAPG) and ligand dependent nuclear receptor corepressor-like protein (LCORL) are positional candidate genes within this region. Previously identified genetic markers in both genes were associated with average daily gain (ADG) and average daily feed intake (ADFI) in a crossbred population of beef steers. These markers were also associated with hot carcass weight, ribeye area and adjusted fat thickness suggesting that they may have a role in lean muscle growth and/or fat deposition. The purpose of this study was to determine whether the transcript abundance of either of these genes in cattle adipose and muscle tissue was associated with variation in feed intake and average daily gain phenotypes. Transcript abundance for NCAPG and LCORL in adipose and muscle tissue was measured in heifers (adipose only), cows and steers using real-time polymerase chain reaction. In the adipose tissue from cows and heifers, a negative correlation between LCORL transcript abundance and ADFI were detected (P = 0.05). In the muscle tissue from cows, transcript abundance of NCAPG was associated with ADG (r = 0.26; P = 0.009). A positive correlation between LCORL transcript abundance from muscle tissue of steers and ADFI was detected (P = 0.04). LCORL protein levels in the muscle of steers were investigated and were associated with ADFI (P = 0.01). These data support our earlier genetic associations with ADFI and ADG within this region and represent the potential for biological activity of these genes in the muscle and adipose tissues of beef cattle; however, they also suggest that sex, age and/or nutrition-specific interactions may affect the expression of NCAPG and LCORL in these tissues.

Lysozyme as an alternative to antibiotics improves growth performance and small intestinal morphology in nursery pigs.

Lysozyme is a 1,4-β-N-acetylmuramidase that has antimicrobial properties. The objective of this experiment was to determine if lysozyme in nursery diets improved growth performance and gastrointestinal health of pigs weaned from the sow at 24 d of age. Two replicates of 96 pigs (192 total; 96 males, 96 females) were weaned from the sow at 24 d of age, blocked by BW and gender, and then assigned to 1 of 24 pens (4 pigs/pen). Each block was randomly assigned 1 of 3 dietary treatments for 28 d: control (two 14-d phases), control + antibiotics (carbadox/copper sulfate), or control + lysozyme (100 mg/kg diet). Pigs were weighed and blood sampled on d 0, 14, and 28 of treatment. Blood was analyzed for plasma urea nitrogen (PUN) and IgA. At 28 d, pigs were killed, and samples of jejunum and ileum were collected and fixed for intestinal morphology measurements. An additional jejunum sample was taken from the 12 pigs with the median BW per treatment to determine transepithelial electrical resistance (TER). Pigs consuming antibiotics or lysozyme grew at a faster rate than control pigs (0.433 ± 0.009 and 0.421 ± 0.008 vs. 0.398 ± 0.008 kg/d, respectively; P < 0.03), which resulted in heavier ending BW (20.00 ± 0.31, 19.8 ± 0.29, and 18.83 ± 0.32 kg, respectively; P < 0.03). Feed intake was not different (P > 0.48), but G:F was improved in pigs consuming antibiotics or lysozyme (0.756 ± 0.014, 0.750 ± 0.021, and 0.695 ± 0.019 kg/kg; P < 0.05). Immunoglobulin A (P < 0.03) and PUN (P < 0.01) increased during the experiment, regardless of dietary treatment (P > 0.48). Dietary treatment did not affect TER (P > 0.37), but gilts had lower TER compared with barrows (P < 0.04). No differences in villi height or crypt depth were observed in the ileum (P > 0.53). However, jejunum villi height was increased and crypt depth was decreased in pigs consuming antibiotics or lysozyme (P < 0.001), resulting in an increased villi height:crypt depth of 72% (P < 0.001). Thus, we concluded that lysozyme is a suitable alternative to carbadox/copper sulfate diets fed to pigs weaned from the sow at 24 d of age.

Granulated lysozyme as an alternative to antibiotics improves growth performance and small intestinal morphology of 10-day-old pigs.

Lysozyme is a 1,4-β-N-acetylmuramidase that has antimicrobial properties. The objective of this experiment was to determine the effect of a purified granulated lysozyme, compared with antibiotics, on growth performance, small intestinal morphology, and Campylobacter shedding in 10-d-old pigs. Forty-eight pigs (n = 16 per treatment), with an initial BW of 4.0 ± 0.1 kg (P > 0.40), were weaned at 10 d of age, blocked by litter and sex, and assigned to pens (8 pigs/pen). Each block was randomly assigned to consume 1 of 3 liquid dietary treatments for 14 d: a control diet, the control diet + lysozyme (100 mg/kg of diet), or the control diet + antibiotics (neomycin and oxytetracycline, 16 mg/kg of diet). Pigs were weighed and blood was sampled on d 0, 7, and 14. Blood was analyzed for plasma urea N and IgA. After 14 d of treatment, pigs were killed and samples of the jejunum and ileum were collected and fixed to measure villus height and crypt depth. Rectal swabs were taken on d 0, 7, and 14 of treatment, and samples of ileal and cecal contents were taken at d 14 of treatment to determine the presence of Campylobacter. Pigs consuming lysozyme and antibiotics gained BW at a faster rate than did control pigs over the course of the study (402 ± 12 and 422 ± 14 g/d, respectively, vs. 364 ± 14 g/d; P < 0.02), resulting in heavier ending BW (9.9 ± 0.3, 9.9 ± 0.3, and 9.0 ± 0.2 kg for pigs in the lysozyme, antibiotic, and control groups, respectively; P < 0.03). Immunoglobulin A decreased and plasma urea N increased over the course of the study (P < 0.1), regardless of dietary treatment (P > 0.6). Crypt depth was increased in pigs fed lysozyme- and antibiotic-treated diets, compared with pigs fed the control diet, in both the jejunum (60.0 ± 2.8 and 62.2 ± 3.0 µm, respectively, vs. 50.7 ± 3.1 µm; P < 0.03) and ileum (76.0 ± 7.5 and 72.2 ± 5.0 µm, respectively, vs. 52.4 ± 3.5 µm; P < 0.02). Villus height did not differ in the jejunum (P > 0.2) but was increased in the ileum of pigs consuming the lysozyme- and antibiotic-treated diets, compared with pigs fed the control diet (312 ± 20 and 314 ± 10 µm, respectively, vs. 263 ± 15 µm; P < 0.4). Small intestinal total mucosa and mucosal protein concentrations, as well as disaccharidase-specific activities, were not altered by lysozyme or antibiotics (P > 0.05). Campylobacter was detected in 27% of control samples but in only 5% of samples from pigs fed antibiotics and 8% of samples from pigs fed lysozyme (P < 0.01). Thus, granulated lysozyme is a suitable alternative to antibiotics for 10-d-old pigs consuming manufactured liquid diets.

Transcriptome differences in the rumen of beef steers with variation in feed intake and gain

BACKGROUND Feed intake and gain are economically important traits in beef production. The rumen wall interacts with feed, microbial populations, and fermentation products important to cattle nutrition. As such, it is likely to be a critical component in the beef steers ability to utilize feedstuffs efficiently. To identify genes associated with steer feed intake and body weight gain traits, and to gain an understanding of molecules and pathways involved in feed intake and utilization, RNA sequencing (RNA-Seq) was performed on rumen papillae from 16 steers with variation in gain and feed intake. Four steers were chosen from each of the four Cartesian quadrants for gain×feed intake and used to generate individual RNA-Seq libraries. RESULTS Normalized read counts from all of the mapped reads from each of the four groups of animals were individually compared to the other three groups. In addition, differentially expressed genes (DEGs) between animals with high and low gain, as well as high and low intake were also evaluated. A total of 931 genes were differentially expressed in the analyses of the individual groups. Eighty-nine genes were differentially expressed between high and low gain animals; and sixty-nine were differentially expressed in high versus low intake animals. Several of the genes identified in this study have been previously associated with feed efficiency. Among those are KLK10, IRX3, COL1A1, CRELD2, HDAC10, IFITM3, and VIM. CONCLUSIONS Many of the genes identified in this study are involved with immune function, inflammation, apoptosis, cell growth/proliferation, nutrient transport, and metabolic pathways and may be important predictors of feed intake and gain in beef cattle.

Lysozyme as an alternative to antibiotics improves performance in nursery pigs during an indirect immune challenge

Lysozyme is a 1,4-β-N-acetylmuramidase that has antimicrobial properties. The objective of this study was to determine the effect of lysozyme and antibiotics on growth performance and immune response during an indirect immune challenge. Two replicates of 600 pigs each were weaned from the sow at 26 d of age, blocked by litter and sex, and then randomly assigned to 1 of 24 pens in either a nursery room that had been fully disinfected or a nursery room left unclean since the previous group of pigs. Within a room, pigs were randomly assigned to either control diets (2 phase nursery regime), control diets + antibiotics (chlortetracycline/tiamulin hydrogen fumarate), or control diets + lysozyme (100 mg/kg diet). Pig weights and feed disappearance were measured and blood was collected on d 0, 14, and 28 of treatment. A group of 20 pigs were killed at 24 d of age for initial body composition analysis and 10 pigs of median weight were killed per diet room combination for body composition analysis after 28 d of treatment. Control + antibiotics and control + lysozyme-fed pigs grew at a faster rate for the 28-d study compared to control pigs (318 ± 14,320 ± 15 vs. 288 ± 15 g/d, respectively; P < 0.05), regardless of nursery environment (P > 0.05). The indirect immune challenge did not alter growth performance from d 0 to 14 of treatment but decreased ADG from d 14 to 28 of the study (415 ± 15 vs. 445 ± 13 g/d ;: P < 0.05). Feed intake was not altered by the nursery environment (P > 0.61) or dietary treatments (P > 0.10), but feed efficiency was worsened by the indirect immune challenge (P < 0.05) and improved by both control + antibiotics and control + lysozyme diets (P < 0.01). The immune challenge did not alter nutrient accretion (P > 0.25), but both control + antibiotics and control + lysozyme pigs had decreased accretion of whole-body lipid (P < 0.01) and increased accretion of protein (P < 0.09). Blood levels of tumor necrosis factor-α (TNF-α; P < 0.01), haptoglobin (P < 0.09), and C-reactive protein (CRP; P < 0.01) were higher due to the indirect immune challenge compared to pigs reared in the clean nursery (P < 0.05). In addition, pigs consuming antibiotics or lysozyme had lower TNF-α, haptoglobin, and CRP compared to control pigs, regardless of nursery environment (P < 0.04). Thus, lysozyme is a suitable alternative to antibiotics in swine nursery diets, and lysozyme ameliorates the effects of a chronic indirect immune challenge.

Effects of antimicrobials fed as dietary growth promoters on faecal shedding of Campylobacter, Salmonella and shiga-toxin producing Escherichia coli in swine.

To determine whether antimicrobials commonly used in swine diets affect zoonotic pathogen shedding in faeces.

The effects of Capn1 gene inactivation on skeletal muscle growth, development, and atrophy, and the compensatory role of other proteolytic systems.

Myofibrillar protein turnover is a key component of muscle growth and degeneration, requiring proteolytic enzymes to degrade the skeletal muscle proteins. The objective of this study was to investigate the role of the calpain proteolytic system in muscle growth development using μ-calpain knockout (KO) mice in comparison with control wild-type (WT) mice, and evaluate the subsequent effects of silencing this gene on other proteolytic systems. No differences in muscle development between genotypes were observed during the early stages of growth due to the up regulation of other proteolytic systems. The KO mice showed significantly greater m-calpain protein abundance (P < 0.01) and activity (P < 0.001), and greater caspase 3/7 activity (P < 0.05). At 30 wk of age, KO mice showed increased protein:DNA (P < 0.05) and RNA:DNA ratios (P < 0.01), greater protein content (P < 0.01) at the expense of lipid deposition (P < 0.05), and an increase in size and number of fast-twitch glycolytic muscle fibers (P < 0.05), suggesting that KO mice exhibit an increased capacity to accumulate and maintain protein in their skeletal muscle. Also, expression of proteins associated with muscle regeneration (neural cell adhesion molecule and myoD) were both reduced in the mature KO mice (P < 0.05 and P < 0.01, respectively), indicating less muscle regeneration and, therefore, less muscle damage. These findings indicate the concerted action of proteolytic systems to ensure muscle protein homeostasis in vivo. Furthermore, these data contribute to the existing evidence of the importance of the calpain systems involvement in muscle growth, development, and atrophy. Collectively, these data suggest that there are opportunities to target the calpain system to promote the growth and/or restoration of skeletal muscle mass.

Differences in transcript abundance of genes on BTA15 located within a region associated with gain in beef steers.

Using results from a previous GWAS, we chose to evaluate seven genes located within a 229Kb region on BTA15 for variation in RNA transcript abundance in a library of tissue samples that included adipose, liver, rumen papillae, spleen, muscle, and small intestine epithelial layers from the duodenum, ileum and jejunum collected from steers (n = 14) with positive and negative residual GN near mean dry matter intake (DMI). The genes evaluated were two olfactory receptor-like genes (LOC525033 and LOC618173), RRM1, STIM1, RHOG, PGAP2, and NUP98. The rumen papillae transcript abundance of RHOG was positively correlated with residual GN (P = 0.02) and ruminal STIM1 exhibited a trend towards an association with residual GN (P = 0.08). The transcript abundance of one olfactory receptor (LOC618173) in the ileum was also positively associated with residual GN (P = 0.02) and PGAP2 and LOC525033 in the ileum displayed trends for association with GN (P ≤ 0.1). To further evaluate the differential expression detected in the ileum and rumen of these animals, the transcript abundance of STIM1 and RHOG in the rumen and of PGAP2 and the olfactory receptors in the ileum were assessed in an additional group of 32 animals with divergent average daily gain (ADG) and average daily feed intake (ADFI) collected over two groups. The olfactory receptor, LOC525033, was not expressed in the ileum for the majority of these animals. Only RHOG showed a slight, but non-significant trend towards greater expression in animals with greater gain. We have detected differences in the transcript abundance of genes within this region in the rumen and ileum of animals selected for greater and less residual gain; however, we were unable to validate the expression of these genes in the larger group of cattle possibly due to the differences in phenotype or contemporary group.

Relationship of neuropeptide FF receptors with pubertal maturation of gilts

Abstract Mechanisms governing the timing of puberty in pigs are poorly understood. A genome-wide association study for age at first estrus in pigs identified candidate genes including neuropeptide FF receptor 2 (NPFFR2), which is a putative receptor for RFamide-related peptides (RFRP). RFRP has been shown to negatively regulate secretion of reproductive hormones from hypothalamic and pituitary tissue of pigs in culture. Here, the porcine NPFFR2 gene was further screened and four potentially functional variants were identified to be associated with age at first estrus in pigs (1,288 gilts). The RFRP neurons in the porcine hypothalamus were localized in the paraventricular and dorsomedial nuclei with RFRP fibers in the lateral hypothalamic area. There were marked changes in expression of NPFF receptors in the anterior pituitary gland and hypothalamus of gilts beginning with the peripubertal period. The hypothesis that NPFF receptor function is related to secretion of luteinizing hormone (LH) in gilts was tested with various NPFF receptor ligands. The NPFF receptor antagonist RF9 stimulated a pulse-like release of LH in prepubertal gilts. The putative NPFF receptor agonist RFRP3modestly suppressed LH pulses in ovariectomized (OVX) prepubertal gilts. A porcine-specific RFRP2 failed to have an effect on LH secretion in OVX prepubertal gilts despite its high degree of homology to avian gonadotropin-inhibitory hormone. Results indicate that an RFRP system is present in the pig and that NPFFR2 is important for pubertal onset in gilts. It is not clear if this regulation involves major control of LH secretion or another unknown mechanism. Summary Sentence Neuropeptide FF receptors play a role in attainment of puberty in pigs.

Effect of lysozyme or antibiotics on faecal zoonotic pathogens in nursery pigs

The objective of this study was to determine the effect of lysozyme and antibiotics on zoonotic pathogen shedding in faeces from nursery pigs housed without and with an indirect disease challenge.