Maternal influences on beef calf rumen microbiome in the first 4 wk of life.

The preruminant microbiome has the potential to set the stage for later life feed efficiency and is critical to proper development within the rumen. We hypothesized that the rumen microbiome is established at or near birth and is subject to maternal influences that can influence preruminant and postruminant microbial profiles. Our objective was to determine how mode of delivery and rearing affected the development of the rumen microbiome. Bred mature Charolais cows were randomly allocated to one of the three treatment groups: control (CON; n = 8), bottle reared (BOT; n = 8), and caesarian section (CSET; n = 8), where CON was vaginal birth and raised by their dam; BOT was vaginal birth, then removed 24-h post-parturition, and raised on commercial milk replacer; and CSET was born via caesarian section and raised by their respective dams. Calf rumen fluid was collected from calves at 1, 3, and 28 d of age via oral lavage and metagenomic shotgun sequencing was performed using the Illumina NextSeq 500 platform. Sequence data were analyzed utilizing Metataxa2 for taxonomic assignment followed by QIIME to determine α- and ß-diversity differences. A total of 1,113 taxa had differential abundance when comparing day while 66 taxa had differential abundance across treatment groups. There were no differences across treatment group richness (P > 0.05), but day 28 was significantly more rich (P = 0.003) compared with days 1 and 3 with no difference between days 1 and 3 (P = 0.58). No differences in ß-diversity were detected across treatment group with the exception of greater variance in the BOT and CSET compared with the CON (P = 0.048). Microbial profiles of day 1 are more similar to each other than day 3 or 28 (P = 0.03); day 3 is more similar to each other than day 1 or 28 (P = 0.03); and day 28 is more similar to each other than day 1 or 3 (P = 0.03). These data suggest that while treatment group did not have a large impact on microbial diversity, several specific taxa were affected by treatment group. Day affects the microbial diversity both within and among samples. Understanding how these profiles shift with age is critical to understanding key intervention periods for optimal alteration of the microbiome.

Microarray analysis of subcutaneous adipose tissue from mature cows with divergent body weight gain after feed restriction and realimentation.

Body weight response to periods of feed restriction and realimentation is critical and relevant to the agricultural industry. The purpose of this study was to evaluate differentially expressed genes identified in subcutaneous adipose tissue collected from cows divergent in body weight (BW) gain after feed restriction and realimentation. We compared adipose samples from cows with greater gain based on average daily gain (ADG) during realimentation with samples from cows with lesser gain. Specifically, there were four comparisons including two comparing the high and low gain animals across each feeding period (feed restriction and realimentation) and two that compared differences in feed restriction and realimentation across high or low gain classifications. Using microarray analysis, we provide a set of differentially expressed genes identified between the high and low gain at both periods of nutrient restriction and realimentation. These data identify multiple differentially expressed genes between these two phenotypes across both nutritional environments.

Relationships between the genes expressed in the mesenteric adipose tissue of beef cattle and feed intake and gain.

Mesenteric fat, a depot within the visceral fat, accumulates in cattle during maturation and finishing and may be a potential source of production inefficiency. The aim of this study was to determine whether the genes expressed in the mesenteric fat of steers were associated with body weight gain and feed intake. Sixteen steers chosen by their rank of distance from the bivariate mean for gain and feed intake were used for this study. Mesenteric fat was obtained and evaluated for differences in gene expression. A total of 1831 genes were identified as differentially expressed among steers with variation in feed intake and gain. Many of these genes were involved with metabolic processes such as proteolysis, transcription and translation. In addition, the Gene Ontology annotations including transport and localization were both over-represented among the differentially expressed genes. Pathway analysis was also performed on the differentially expressed genes. The superoxide radical degradation pathway was identified as over-represented based on the differential expression of the genes GPX7, SOD2 and TYRP1, suggesting a potential role for oxidative stress or inflammatory pathways among low gain-high intake animals. GPX7 and SOD2 were in lower transcript abundance, and TYRP1 was higher in transcript abundance among the low gain-high feed intake animals. The retinoate biosynthesis pathway was also enriched due to the differential expression of the genes AKR1C3, ALDH8A1, RDH8, RDH13 and SDR9C7. These genes were all more highly expressed in the low gain-high intake animals. The glycerol degradation and granzyme A signaling pathways were both associated with gain. Three glycerol kinase genes and the GZMA gene were differentially expressed among high vs. low gain animals. Mesenteric fat is a metabolically active tissue, and in this study, genes involved in proteolysis, transcription, translation, transport immune function, glycerol degradation and oxidative stress were differentially expressed among beef steers with variation in body weight gain and feed intake.

Effect of abomasal butyrate infusion on gene expression in the duodenum of lambs.

A previous study infusing butyrate into the abomasum of sheep produced increased oxygen, glucose, glutamate, and glutamine uptake by the portal-drained viscera. These changes were thought to be partially due to increases in glycolysis and cell proliferation. The purpose of this study was to evaluate the duodenum transcriptome of control and butyrate-treated lambs to determine whether genes involved in these pathways were altered. Polled Dorset lambs ( = 9) received a pulse dose of either butyrate (10 mg/kg BW) or an equal volume of a buffered saline solution (1 mL/kg BW) daily at the time of feeding. Lambs were euthanized approximately 4 h after treatment/feeding on d 21, and a sample of duodenal mucosa was obtained from which total RNA was isolated for microarray analysis. A total of 230 genes were differentially expressed ( < 0.05). Pathway analyses performed with the differentially expressed genes revealed glycolysis, fatty acid activation/biosynthesis, UDP-N-acetyl-ᴅ-galactosamine biosynthesis, γ-Linolenate biosynthesis, and mitochondrial ʟ-carnitine shuttle pathways up-regulated by the butyrate treatment. Additionally, expression of functional gene clusters related to mitochondrial function was found to be enriched ( < 0.05) with the butyrate treatment. These data could partially explain the metabolite flux changes that were observed with the butyrate treatment; specifically the increase in glucose uptake and glycolysis pathway upregulation and the increased oxygen uptake and upregulation of mitochondria function-related genes.

Effects of high-sulphur water on hepatic gene expression of steers fed fibre-based diets.

Sulphur-induced polioencephalomalacia (sPEM), a neurological disorder affecting ruminants, is frequently associated with the consumption of high-sulphur (S) water and subsequent poor performance. Currently, there is no economical method for S removal from surface water sources, and alternative water sources are typically neither readily available nor cost-effective. Determination of genes differentially expressed in response to high-S water consumption may provide a better understanding of the physiology corresponding to high dietary S and ultimately lead to the development of treatment and prevention strategies. The objective of this study was to determine changes in gene expression in the liver, an organ important for S metabolism, of fibre-fed steers consuming high-S water. For this study, liver tissues were collected on the final day of a trial from yearling steers randomly assigned to low-S water control (566 mg/kg SO4 ; n = 24), high-S water (3651 mg/kg SO4 ; n = 24) or high-S water plus clinoptilolite supplemented at either 2.5% (n = 24) or 5.0% (n = 24) of diet dry matter (DM). Microarray analyses on randomly selected healthy low-S control (n = 4) and high-S (n = 4; no clinoptilolite) steers using the Affymetrix GeneChip Bovine Genome Array revealed 488 genes upregulated (p < 0.05) and 154 genes downregulated (p < 0.05) in response to the high- vs. low-S water consumption. Real-time RT-PCR confirmed the upregulation (p < 0.10) of seven genes involved in inflammatory response and immune functions. Changes in such genes suggest that ruminant animals administered high-S water may be undergoing an inflammation or immune response, even if signs of sPEM or compromised health are not readily observed. Further study of these, and other affected genes, may deliver new insights into the physiology underlying the response to high dietary S, ultimately leading to the development of treatments for high S-affected ruminant livestock.

Selection for placental efficiency in swine: conceptus development.

The objective of this study was to evaluate correlated responses in conceptus development and traits physiologically relevant for placental function in swine from a selection experiment that resulted in differences in placental weight (PW) and efficiency (PE = birthweight/placental weight). Generation 3, second parity females from 2 lines with a history of selection on an index predicting either high PE (HPE) or low PE (LPE) were mated within line to produce Generation 4 litters for evaluation at d 30, 50, 70, 90, and 110 of gestation (n = 5/line × d combination). Maternal and fetal traits were analyzed by using a model including the fixed effects of line and gestational age, and the random effect of sire within line. Uterine length was not different between lines at any gestational age, but increased (P = 0.06) from 275.0 ± 23.1 cm at d 30 to 338.3 ± 23.3 cm at d 50, and remained relatively unchanged to d 110. Fetal weight was not different between lines from d 30 to 90, but was less (P = 0.02) in HPE than LPE at d 110 (1,280.6 ± 77.0 vs. 1,551.1 ± 75.3 g, respectively). Crown-rump length was not different between lines from d 30 to 70, but tended (P = 0.09) to be longer in HPE than LPE at d 90 (265.8 ± 8.8 vs. 241.2 ± 10.6 mm, respectively) and was shorter (P = 0.04) in HPE than LPE at d 110 (290.6 ± 5.0 vs. 304.9 ± 4.5 mm, respectively). Placental weight increased in both lines from d 30 to 50, at which point it remained relatively unchanged through the rest of pregnancy, except in LPE that showed a second increase from d 90 to 110. As a result, placental weight was not different between lines from d 30 to 90, but was less (P < 0.01) in HPE than LPE at d 110 (244.6 ± 32.3 vs. 379.2 ± 24.5 g, respectively). Line differences in placental efficiency were not significant at any gestational age. Implantation site length increased slowly for both lines from d 30 to 90, where it remained unchanged to d 110. Implantation site area was greater (P < 0.05) in HPE than LPE at d 30 and 50, but was not different between lines for the remainder of pregnancy. These results suggest that in Western breeds, a reduction in placental weight through selection is not accompanied by compensation in placental nutrient transfer and may result in decreased prenatal survival.

Effects of supplemental molybdenum on animal performance, liver copper concentrations, ruminal hydrogen sulfide concentrations, and the appearance of sulfur and molybdenum toxicity in steers receiving fiber-based diets.

Poor performance and S-induced polioencephalomalacia (sPEM) have been observed in ruminant livestock in high-S drinking water regions. No gainful method of removing S from drinking water is available and therefore a feed supplement that negates the effects of high-S water is needed. Our objective was to determine if supplementing Mo improves health and performance of steers administered a high-fiber diet and high-S drinking water. We hypothesized that if the supplemental Mo adequately bound excess S in the rumen, it would not be available at toxic concentrations. Yearling steers (n = 96; 260.0 ± 1.3 kg BW) were stratified by pretrial BW into 12 feedlot pens (n = 8 steers per pen). One of 3 treatments, low-S water (LS; 375 mg SO(4)/L), high-S water (HS; 2,218 mg SO(4)/L), or high-S water plus Mo (HSMO; 2,218 mg SO(4)/L; 187.5 mg Mo/kg DM), were randomly assigned to pens within 4 blocks for a 56-d trial. Body weights were recorded on d -2, -1, 29, 56, and 57, ruminal H(2)S concentrations were measured by rumenocentesis on d -1, 29, and 57, and liver biopsies were performed on d -1 and 57. Performance data were analyzed over the 56-d trial period (overall) as well as over 2 periods: Period 1 (d 0 to d 28) and Period 2 (d 29 to d 56). One case of sPEM was confirmed by the presence of cortical lesions in the HS treatment group. Daily DMI and ADG were affected by treatment and period (P < 0.001) main effects. The LS steers had the greatest (P < 0.05) DMI followed by HS and HSMO steers, respectively. Similar results were observed for ADG. Daily water intake was affected (P < 0.001) by period only, with greater daily water intake in Period 2 than Period 1. Change in hepatic concentrations of Cu, Fe, and Mo over the course of the trial were all affected (P < 0.001) by treatment. Hepatic Cu increased from d 1 to 57 in LS and HS steers but was depleted in HSMO steers. Hepatic Fe and Mo increased in HSMO steers only. Ruminal H(2)S concentrations were affected by treatment (P < 0.021), with greater H(2)S concentrations in HSMO compared with LS and HS steers. Signs of Mo toxicity such as severe diarrhea, loss of body condition, anorexia, changes in hair color, and stiffness in joints were observed in the Mo supplemented steers. These results indicate that added dietary Mo does not adequately bind excess S in the rumen, causing aggravated toxic effects from potentially both the high dietary S and Mo.

Renin mRNA is upregulated in testes and testicular cells in response to treatment with aflatoxin B1.

Aflatoxin B(1) (AFB(1)) has been shown to affect fertility in many species; however, the exact molecular mechanisms associated with the disruption are not known. Our objectives were to determine changes in testicular gene expression due to exposure to AFB(1) and to investigate which cell types were affected by treatment with AFB(1). Male mice 4 wk of age were administered a daily placebo (control; N = 9) or 50 µg/kg AFB(1) (AFB(1) treated; N = 10) daily for 45 days. Males were then mated to four females each for 8 days. Male mice were characterized as being "Tolerant" (N = 3) or "Intolerant" (N = 3) to the effects of AFB(1) based on positive terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining in the testes and the number of pups sired. Tolerant males produced a similar average number of fetuses (mean ± SEM) (12.5 ± 1.2) per male as selected control males (13.4 ± 1.2), but more fetuses (P = 0.01) than Intolerant males (7.6 ± 1.2). The number of terminal deoxynucleotidyl transferase dUTP nick end labeling-positive cells in Intolerant males tended to be (P = 0.10) greater (136.5 ± 27.2) than in Tolerant (55.0 ± 22.2) and selected control (54.3 ± 22.2) males. Affymetrix microarray (Sunnyvale, CA, USA) analysis revealed differential expression (P < 0.05) of 193 extra cellular space and signaling genes, 49 signal transduction genes, 45 immune regulation genes, and 230 cell differentiation genes in the testis. Renin was commonly represented amongst many clusters and was chosen for further analyses. Upregulation (P < 0.001) of Renin in Tolerant mice (N = 3) compared with Intolerant mice (N = 3) was confirmed by real-time reverse transcription polymerase chain reaction (RT-PCR) (P = 0.05). This upregulation (P = 0.01) was also observed in representative AFB(1) treated males (N = 8) compared with control males (N = 8) selected for real-time reverse transcription polymerase chain reaction analysis. Spermatogonia cultured in vitro and treated with 0, 5, 10, or 20 µg/mL AFB(1) (N = 6 per treatment) resulted in a 10-fold upregulation (P = 0.01) of Renin message at the 20 µg/mL level, whereas Leydig tumor cells showed similar differences (P = 0.03) in message for Renin in treated (10 and 20 µg/mL) versus control cell cultures. Based on these results, we inferred a role for Renin at the molecular level in the response to the adverse effects of AFB(1) in male mice.

Camelina meal and crude glycerin as feed supplements for developing replacement beef heifers.

Angus × Gelbvieh rotationally crossbred yearling heifers (n = 99, yr 1; n = 105, yr 2) were used in a 2-yr randomized complete block design experiment with repeated measures to determine the effect of feeding camelina biodiesel coproducts (meal and crude glycerin) on serum concentrations of triiodothyronine, thyroxine, insulin, ß-hydroxybutyrate, and glucose, as well as on growth and reproductive performance. Heifers were assigned to 1 of 15 pens, and pens were assigned initially to receive 7.03 k·â€¢heifer(-1)·d(-1) of bromegrass hay plus 0.95 kg·heifer(-1)·d(-1) of 1 of 3 supplements for 60 d before breeding: 1) control (50% ground corn and 50% soybean meal, as-fed basis); 2) mechanically extracted camelina meal; or 3) crude glycerin (50% soybean meal, 33% ground corn, 15% crude glycerin, 2% corn gluten meal; as-fed basis). Preprandial blood samples were collected via the jugular vein on d 0, 30, and 60 of the feeding period. A 2-injection PGF(2α) protocol (d 60 and 70 of the study) was used to synchronize estrus. Heifers were artificially inseminated 12 h after estrus was first detected. Heifers not detected in estrus within 66 h received a GnRH injection and were artificially inseminated. Dietary treatment × sampling period interactions were not detected (P = 0.17 to 0.87). Dietary treatment did not affect BW (P = 0.44 to 0.59) or serum concentrations of thyroxine (P = 0.96), ß-hydroxybutyrate (P = 0.46), glucose (P = 0.59), or insulin (P = 0.44). Serum concentrations of triiodothyronine were greater (P = 0.05) in heifers fed camelina meal. Additionally, dietary treatment did not affect the percentage of heifers detected in estrus before timed AI (P = 0.83), first-service pregnancy rates of those heifers detected in estrus (P = 0.97), or overall first-service pregnancy rates (P = 0.58). Heifers fed camelina meal, however, had greater (P = 0.05) first-service pregnancy rates to timed AI than did heifers fed the control and crude glycerin supplements. The cost per pregnancy was similar for heifers fed the crude glycerin or the control supplement, whereas the cost per pregnancy was the least for heifers fed camelina meal. We conclude that camelina coproducts can replace conventional corn-soybean meal supplements in the diets of developing replacement beef heifers.

Effects of dietary aflatoxin on the hepatic expression of apoptosis genes in growing barrows.

The most common and toxic form of aflatoxin, aflatoxin B(1) (AFB(1)), is produced by molds growing on crops. Use of moldy corn can result in high concentrations of AFB(1) in swine diets, which could potentially lead to an increased incidence of aflatoxicosis, a disease associated with decreased health and performance through reduced feed intake, reduced BW gain, and impaired liver function. The objective of this study was to determine the effects of AFB(1) on the hepatic gene expression of growing barrows. Ninety Duroc × Yorkshire crossbred barrows (age = 35 ± 5 d; initial BW = 14.2 ± 3.0 kg) were allocated to 9 pens with 10 pigs per pen, and randomly assigned in a 3 × 3 factorial arrangements of treatments to receive diets containing 0 µg/kg of AFB(1), 250 µg/kg of AFB(1), or 500 µg/kg of AFB(1) for 7, 28, or 70 d. Because performance was most affected in animals administered AFB(1) for an extended period, liver samples from d 70 animals were used for RNA-sequencing analysis. Of 82,744 sequences probed, 179 had transcripts that were highly correlated (r ≥ |0.8|; P < 0.0001) with treatment. Of the 179 significant transcripts, 46 sequences were negatively and 133 sequences positively related to treatment. Forty-three unique functional groups were identified. Genes within the apoptosis regulation functional group were selected for 1) confirmation of d 70 gene expression differences using real-time reverse-transcription (RT)-PCR (n = 4 genes), and 2) investigation of d 7 expression to identify early responses to AFB(1) (n = 15 genes) using real-time RT-PCR. Expression of the 4 apoptosis genes selected for confirmation, cyclin-dependent kinase inhibitor 1A, zinc finger matrin type 3, kininogen 1, and pim-1 oncogene, was confirmed with real-time RT-PCR. Of the 15 genes tested in d 7 liver samples, 4 were differentially expressed: cyclin-dependent kinase inhibitor 1A; zinc finger matrin type 3; tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta polypeptide; and apoptosis enhancing nuclease. Results from this study demonstrate that administration of an AFB(1)-contaminated diet to growing barrows alters hepatic gene expression, and in particular apoptosis genes.

Hypothalamic expression of genes for appetite regulators and estrogen α, estrogen ß and leptin receptors in obese dams and their fetuses.

Under- and over-nutrition during gestation may influence fetal hypothalamic development resulting in individuals predisposed to adverse health effects. This study examined fetuses from obese and control ewes to determine whether dam obesity alters hypothalamic expression of fetal appetite regulatory genes. A second objective was to contrast the expression of appetite regulatory genes in ewes that become the most obese to those that remained in moderate body condition on the same energy-rich diet. Multiparous, western white-faced ewes were weighed and individually fed 100% (control) or 150% (obese) of National Research Council requirements from day 60 before mating until day 75 of gestation. At day 75 of gestation, fetuses were collected and weighed. Hypothalamic tissue from fetal lambs and dams was collected and frozen for mRNA extraction. Dam obesity (P ≥ 0.16), fetal sex (P ≥ 0.44) or their interaction (P ≥ 0.42) did not affect the relative expression of fetal hypothalamic regulators of appetite, including neuropeptide Y, agouti-related protein, pro-opiomelanocortin, cocaine- and amphetamine-regulated transcript and receptors for leptin. Maternal obesity at day 75 of gestation in ewes did not affect developmental mechanisms responsible for the expression of fetal appetite regulatory genes and would not be expected to predispose offspring to adult-onset obesity through disrupted appetite regulation at this developmental time point. In the ewe, appetite regulatory genes did not differ (P > 0.20) with ewe adiposity; however, expression of estrogen receptor α, but not ß (P = 0.37), in the medial basal hypothalamus was greater (P = 0.04) in obese than in control ewes.

Effects of dietary aflatoxin on the health and performance of growing barrows.

Aflatoxins, especially aflatoxin B1 (AFB1), can be greater in dried distillers grains with solubles (DDGS) because it can be concentrated during the ethanol production process. Increased use of DDGS in swine diets could potentially lead to an increased incidence of aflatoxicosis, a disease associated with decreased feed intake, reduced BW gain, and impaired liver function. The objective of this study was to determine the effects of AFB1 on the health, performance, and serum profile of growing barrows. Ninety Duroc × Yorkshire crossbred barrows were purchased (age = 35 ± 5 d; BW = 14.2 ± 3.0 kg), allocated to 9 pens with 10 pigs per pen, and randomly assigned to receive diets containing 0 µg/kg of AFB1 (CON), 250 µg/kg of AFB1 (LO), or 500 µg/kg of AFB1 (HI) for 7, 28, or 70 d in a 3 × 3 factorial arrangement of treatments. Feed intake was measured daily, and pigs were weighed and blood samples collected weekly. Serum was analyzed for concentrations of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (BILI), and blood urea nitrogen (BUN). Both ADFI and ADG were negatively affected (P ≤ 0.001) by AFB1 treatment. Average daily feed intake was less (P < 0.05) in HI barrows than in CON barrows from wk 5 to 10 and was less (P < 0.05) in LO barrows than in CON barrows in wk 5 and again from wk 8 to 10. Also, ADFI was less (P = 0.022) in HI barrows than LO barrows in wk 10. Decreased ADG (P < 0.05) was observed in HI barrows than in CON barrows in wk 8 and 10; no differences (P ≥ 0.665) in ADG were noted between CON and LO barrows. There was no effect (P ≥ 0.080) of AFB1 treatment on ALT or BILI concentrations. However, both AST and BUN were affected (P < 0.05) by AFB1 treatment. Serum AST was greater (P = 0.010) in LO barrows than CON barrows in wk 5, and serum BUN was greater (P = 0.004) in CON barrows than LO barrows in wk 3. Results from this study demonstrate that the performance and health of young growing barrows were affected by consumption of an AFB1-contaminated diet, especially when fed for a more extended period.

Differential gene expression of ewes varying in tolerance to dietary nitrate.

Ruminants consuming diets with increased concentrations of nitrate (NO(3)(-)) can accumulate nitrite (NO(2)(-)) in the blood, resulting in toxicity. In a previous experiment, ewes identified as highly tolerant to subacute dietary NO(3)(-) were able to consume greater amounts of NO(3)(-) than lowly tolerant ewes without exhibiting signs of toxicity. We hypothesized that highly tolerant and lowly tolerant ewes differ in their ability to metabolize NO(3)(-) and thereby differ in the expression of hepatic genes involved in NO(3)(-) metabolism. Therefore, our objective was to identify hepatic genes differentially expressed between ewes classified as lowly tolerant and highly tolerant after administration of a subacute quantity of dietary NO(3)(-). Analysis of the Bovine Oligonucleotide Microarray data identified 100 oligonucleotides as differentially expressed (P < 0.05) between lowly tolerant and highly tolerant ewes. Functional analysis of the genes associated with these oligonucleotides revealed 2 response clusters of interest: metabolic and stress. Genes of interest within these 2 clusters (n = 17) and nonclustered genes with the greatest fold changes (FC; n = 5) were selected for validation by real-time reverse-transcription PCR. Relative expression, genomic regulation, and FC agreed between microarray and real-time reverse-transcription-PCR analyses, and FC differences (P < 0.05) between lowly tolerant and highly tolerant ewes were confirmed for 12 genes. Metabolic genes that were downregulated (P ≤ 0.032) in lowly tolerant ewes vs. highly tolerant ewes included aldehyde oxidase 1, argininosuccinate lyase, putative steroid dehydrogenase, 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase1, and sterol carrier protein 2. In contrast, the metabolic gene homeobox was upregulated (P = 0.037) in lowly tolerant ewes. The glutathione peroxidase 3 and inter-α (globulin) inhibitor H4 genes in the stress response cluster were upregulated (P ≤ 0.045) in lowly tolerant ewes. Genes with the greatest FC, but did not cluster within the functional analysis included haptoglobin, which was upregulated (P = 0.024) in lowly tolerant ewes, and fatty acid desaturase 2 and thyroid hormone responsive, both of which were downregulated (P ≤ 0.019) in lowly tolerant ewes. Results from this study indicate that hepatic gene expression differs in ewes identified as lowly tolerant and highly tolerant to increased dietary NO(3)(-).

Effects of high-sulfur water and clinoptilolite on health and growth performance of steers fed forage-based diets.

Sulfur-induced polioencephalomalacia (sPEM), a neurological disorder affecting ruminants, is associated with consumption of diets with increased S (high-S). High-S water is commonly found in many western states and is a major source of dietary S for grazing cattle. Consumption of high-S water has been associated with sPEM and decreased performance. Identification of a feed supplement that would counteract the negative effects of high-S water would decrease the incidence of sPEM and prevent performance reductions in regions with problematic water sources. The objectives of this study were to 1) determine the effects of administering high-S drinking water to forage-fed feedlot steers on health and performance, and 2) determine the effectiveness of clinoptilolite, a clay mineral with increased cation-exchange capacity, in negating the effects of high-S drinking water. Yearling steers (n = 96; 318.2 +/- 2.1 kg of BW) were randomly assigned to 1 of 4 treatments for a 77-d trial period: control with low-S water (566 mg of SO(4)/L), high-S water (3,651 mg of SO(4)/L), or high-S water plus clinoptilolite supplemented at 2.5 or 5.0% of the diet DM. Feed and water consumption were measured daily, and all steers were weighed on d -2, -1, 29, 53, 76, and 77. Plasma samples were collected on d 0, 58, and 77, and liver samples on d 0 and 77. There was a greater (P or= 0.546) in ADG or G:F were observed. Plasma Cu decreased (P = 0.029) to a greater magnitude in high-S water steers than the control steers over the 77-d trial period. Mineral analyses of hepatic tissue from randomly selected healthy steers from each treatment group (n = 10 per treatment) showed an interaction (P

Effect of subacute dietary nitrate on production traits and plasma analytes in Suffolk ewes.

Elevated dietary nitrate (NO3-) is associated with production losses in ruminant livestock, resulting in substantial economic losses incurred by producers. Severe drought, fertilization practices and poorly maintained pastures increase the risk of elevated NO3- intake among cattle and sheep. Nitrate is metabolized to nitrite (NO2-) in the rumen and further reduced to ammonia. Ruminants consuming high dietary NO3- vary in ability to efficiently reduce excess NO2- to ammonia. This leads to methemoglobin formation and ultimately NO3- toxicity signs. Variation in individual tolerance to elevated dietary NO3- can be partially attributed to rate and duration of exposure, rate of elimination, metabolism, species and dose. Our objectives were to confirm and quantify variation in individual tolerance to subacute levels of dietary NO3-, and determine if individuals could be identified as highly or lowly tolerant to elevated dietary NO3- based on production traits, plasma analytes and(or) signs of subacute NO3- toxicity. Purebred Suffolk ewes were administered supplement mixed with tap water (control; n = 8) or potassium nitrate (NO3- treated; 300 mg NO3-/kg BW daily; n = 47) for 8 days. Coefficients of variation (CV) indicated that supplement intake was more variable in NO3- treated ewes (CV = 59.3%) than in control ewes (CV = 13.6%). Among NO3- treated ewes, six ewes highly tolerant and six ewes lowly tolerant to elevated dietary NO3- were identified based on individual performance, NO3- treated supplement intake, and signs of toxicity. Supplement intake was lower (P < 0.0001) in NO3- treated ewes than in control ewes, indicating elevated dietary NO3- influences feed intake. Supplement intake differed (P < 0.0001) between control, highly tolerant and lowly tolerant ewes. Supplement intake of highly and lowly tolerant ewes was 82% and 23%, respectively, of the control ewes' intake. Weight change and plasma concentrations of NO2-, cortisol, glucose and retinol were not different (P 0.38) among control, highly tolerant and lowly tolerant ewes. Plasma urea nitrogen (PUN) levels were not different (P = 0.25) between control and lowly tolerant ewes, but were lower (P = 0.02) in highly tolerant ewes than in control ewes. Furthermore, PUN and NO3- treated supplement intake were highly correlated (0.71; P < 0.0001) in lowly tolerant ewes. These results confirm and quantify variation in response to subacute levels of dietary NO3- and indicate that individuals can be identified as highly or lowly tolerant to elevated dietary NO3- based on their performance and NO3- toxicity signs.

Testicular gene expression in male mice divergent for fertility after heat stress.

Fertility losses in male mice occur approximately 18-28 d after heat stress. The objective of this study was to identify gene expression differences in males highly versus lowly fertile after heat stress. Mature male mice were exposed to heat stress (35+/-1 degrees C; n=50) or thermoneutral (21+/-1 degrees C; n=10) conditions for 24 h (Day 0) and hemicastrated (Day 1) to collect tissue for gene expression analyses. Males were subjected to a mating test from Days 18 to 26 when variation in fertility was anticipated. A fertility index was used to rank heat-stressed males and identify those males resistant and susceptible to heat stress, respectively. Microarray analyses were conducted on testis tissues from control (n=5), heat stress resistant (n=5), and heat stress susceptible (n=5) males, and 225 genes were observed to be differentially expressed (P<0.05), including genes involved in chaperone (Canx, Hspcb1, and Tcp1) and catalytic (Fkpb6, Psma7, and Idh1) activity. Expression patterns of these genes were confirmed using real-time RT-PCR. Male progeny from selected sires were similarly divergent in fertility after heat stress. Testicular expression levels of Canx, Hspcb, and Tcp1 genes were determined in these progeny. Hspcb expression was moderately heritable (0.31+/-0.25); however, expression patterns of Canx and Tcp1 were not heritable.

Genetic variation in fertility of heat-stressed male mice.

The damaging effects of heat stress on male fertility are evident in developing spermatozoa expressed in ejaculates 18-28 days post-stress in mice. Our objectives were to: (1) assess genetic variation in fertility of heat-stressed male mice and (2) determine response to selection for fertility after heat stress in male mice. Mature male mice were exposed to heat stress (35+/-1 degrees C; n=50) or control (21+/-1 degrees C; n=10) conditions for 24h (day 0) and then hemicastrated for tissue collection. Two periods of mating tests followed, period 1 (from days 3 to 11) when no reductions in fertility were anticipated, and period 2 (days 18-26) when variation in fertility was expected. Period 2 pregnant females were sacrificed in late gestation. Males were indexed by multiplying overall mean ovulation rate by pre-implantation survival and number of pregnant period 2 mates. The five highest and five lowest ranking males were identified as heat stress resistant and susceptible, respectively. Resistant males were 61.2units superior in the index, 57.5% greater in pregnancy rate, and 57.6 total fetuses greater than susceptible males. Progeny of resistant sires were superior to progeny of susceptible sires in estimated breeding value by 4.5units for the index, 4.1% for pregnancy rate, and 5.2 fetuses (P<0.0001). Heritability estimates for the index, pregnancy rate, and number of fetuses ranged from 0.09 to 0.13, suggesting male fertility following heat stress is heritable and responds to selection.

Genetic and phenotypic relationships of farrowing and weaning survival to birth and placental weights in pigs.

Data obtained during 4 generations of divergent selection for placental efficiency were used to determine factors influencing survival at farrowing and weaning in litters produced by first-parity females. Data were collected from 193 litters and included records on 2,053 individuals. Farrowing survival (FS) and weaning survival (WS) were considered traits of the piglet and were scored 1 if the individual was alive at a time point or 0 if dead. Estimates of (co)variance components for direct and maternal additive genetic effects for FS and WS were obtained using an animal model and computed with the MTDFREML program. Estimates of direct heritability were 0.16 for FS and 0.18 for WS. Estimates of maternal heritability were 0.14 for FS and 0.10 for WS. Genetic correlation estimates between direct and maternal effects were high and negative for both traits. The direct genetic correlation between FS and WS was 0.92. Variables associated with FS and WS were determined using logistic regression procedures. Birth weight (BRW), placental weight, their interaction, and total born can be used as predictors of survival at farrowing in the absence of estimates of genetic merit for survival. The same model, excluding total number born, was the best model for predicting WS. In the presence of BRW information, placental efficiency did not improve the prediction of survival. While it was clearly disadvantageous for a piglet to be below the litter mean in BRW, being above the mean did not provide a substantial advantage in survival. Results from this analysis suggest that it is possible to select for increased survival at farrowing and at weaning. Information on a piglet's BRW, placental weight, litter average BRW, and deviation from litter average BRW can be used to optimize those values at levels resulting in high survival probability.

Selection for placental efficiency in swine: genetic parameters and trends.

The objectives of this study were to estimate response to divergent selection for an index of placental efficiency in swine, and to evaluate the effect of placental efficiency on litter size. The selection index (SI) included total born (TB), birth weight (BRWT), and placental weight (PW), and was designed to increase in the high line (H) or decrease in the low line (L) the efficiency of the placental function (PE), defined as the ratio BRWT:PW. (Co)variance components were estimated for direct and maternal additive effects by using an animal model with MTDFREML procedures. Estimated breeding values were calculated by using records on individual BRWT (n = 2,111), PW (n = 2,006), PE (n = 1,677), and SI (n = 1,677). Litter traits were evaluated using records on 193 litters. The model included the fixed effects of contemporary group for all traits, with the addition of sex for individual traits and parity for litter traits. Litter was fitted as an uncorrelated random effect for all traits, and TB was used as a linear and quadratic covariate for BRWT, PW, and PE. Direct heritability estimates from single-trait models were 0.03, 0.25, 0.18, 0.11, and 0.08 for BRWT, PW, PE, SI, and TB, respectively. Estimated breeding values were compared between lines by using a model including generation, line within generation, and replicate within line as the error term. Estimates of genetic divergence were 20.7 +/- 2.7 g, 0.24 +/- 0.03, 0.11 +/- 0.02, and 0.07 +/- 0.02 per generation for PW, PE, SI, and TB, respectively (P < 0.01), but divergence was not significant for BRWT. At Generation 4, direct EBV was higher in L than in H for PW (55.9 +/- 8.7 vs. -24.2 +/- 9.5 g, respectively; P < 0.01) and higher in H than in L for PE (0.58 +/- 0.10 vs. -0.35 +/- 0.09 g, respectively; P < 0.01). However, EBV was not different for BRWT, SI, or TB. These results indicate that PW and PE are susceptible to change by genetic selection; however, the correlated response in TB was an unexpected genetic trend toward a higher TB in L of 0.05 +/- 0.01 piglets per generation (P < 0.01).

Estimates of genetic parameters for feed intake, feeding behavior, and daily gain in composite ram lambs.

Our objective was to estimate genetic parameters for feed intake, feeding behavior, and ADG in composite ram lambs ((1/2) Columbia, (1/4) Hampshire, (1/4) Suffolk). Data were collected from 1986 to 1997 on 1,239 ram lambs from approximately 11 to 17 wk of age at the U.S. Meat Animal Research Center near Clay Center, NE. Feeding equipment consisted of an elevated pen with an entrance chute that permitted access to the feeder by only one ram lamb at a time, with disappearance of feed measured by an electronic weighing system. Ram lambs were grouped 11 per pen from 1986 to 1989, and nine per pen from 1990 to 1997. Data were edited to exclude invalid feeding events, and approximately 80% of the data remained after edits were applied. Traits analyzed were daily feed intake (DFI), event feed intake (EFI), residual feed intake (RFI), daily feeding time (DFT), event feeding time (EFT), number of daily feeding events (DFE), and ADG. Feed intake traits of DFI and EFI had estimated heritabilities of 0.25 and 0.33, respectively, whereas estimated heritability of RFI was 0.11. Heritability estimates for feeding behavior traits, including DFT, EFT, and DFE, ranged from 0.29 to 0.36. Average daily gain had an estimated heritability of 0.26. Genetic correlations were positive between all pairs of traits, except for RFI and ADG, and that estimate was essentially zero. Phenotypic correlations were generally similar to genetic correlations. Genetic correlations were large (0.80) between DFI and ADG, intermediate between DFI and RFI (0.61) and between DFT and DFE (0.55), and low (0.17 to 0.31) for the other pairs of traits, with the exception of RFI and ADG (-0.03). Genetic correlations between behavioral traits were greater than correlations between behavioral traits and measures of feed intake or ADG; however, selection for ADG and/or feed intake would be expected to cause some changes in feeding behavior.