Negative energy balance in dairy cows is associated with specific changes in IGF-binding protein expression in the oviduct

Negative energy balance (NEB) during early lactation in dairy cows leads to an altered metabolic state that has major effects on the production of IGF family members. Low IGF-I concentrations are associated with poor fertility and therefore we aimed to determine whether NEB exerts a direct effect on IGF expression in the postpartum oviduct. Multiparous Holstein cows were allocated to two treatments (each n=6) designed using differential feeding and milking regimes to produce either mild NEB (MNEB) or severe NEB (SNEB). Animals were slaughtered in week 2 of lactation when divergent metabolic profiles were evident. Oviducts were collected for RNA analysis by real-time RT-PCR and in situ hybridisation. Quantitative measures in oviduct gene expression were obtained for all members of the IGF family (IGF-I/II, IGF-binding proteins (IGFBP) 1-6 and receptors for IGF types 1 and 2), insulin A/B, GH, glucocorticoid and oestrogen alpha/beta. Expression of IGFBP-2 and IGFBP-6 (both of which have a high affinity for IGF-II) was decreased in SNEB relative to MNEB (P<0.05). No other gene was altered by NEB, but IGF-II, IGFBP-3, IGFBP-5 and IGFBP-6 all showed differential expression in different regions of the oviduct. These results indicate that, in addition to low circulating IGF-I after calving, NEB may also influence IGF availability in the oviduct indirectly through changes in specific IGFBP expression. It is possible that the predicted increased signalling by IGF-II may perturb embryo development, contributing to the high rates of embryonic mortality in dairy cows.     

Interrelationships between energy balance and postpartum reproductive function in dairy cattle

Genetic improvement of dairy cows has markedly increased milk yield over the last three decades. Increased production has been associated with reduced conception rates (66% in 1951 versus 40 to 50% since 1975). Because conception rate in dairy heifers has remained higher, the metabolic demands of higher production may be related to the decline in reproductive performance in cows. During early lactation, increasing dietary intake fails to keep pace with rising milk production. The resultant negative energy balance and rate of mobilization of body reserves appear directly related to the postpartum interval to first ovulation and lower conception rate. Delays in the onset of normal ovarian activity, thus limiting the number of estrous cycles before breeding, may account for the observed decrease in fertility. Negative energy balance probably acts similarly to undernutrition and may manifest in delayed ovarian activity by impinging on pulsatile secretion of LH. Lower availability of glucose and insulin may also decrease LH pulsatility or limit ovarian responsiveness to gonadotropins. Alternatively, release of endogenous opioids in association with increasing feed intake or other lactational hormone responses may provide neural or pituitary inhibition of the pulsatile LH production that is requisite for ovarian follicular development.     

Evidence that lamprey GnRH-III does not release FSH selectively in cattle

Experiments were performed to test the hypothesis that lamprey GnRH-III (lGnRH-III) selectively releases FSH. Primary cultures of bovine adenohypophyseal cells were treated with mammalian GnRH (mGnRH) and lGnRH-III (10(-9), 10(-8), 10(-7) and 10(-6) M) or control media in Experiment 1. All doses of mGnRH and the two highest doses of lGnRH-III stimulated (P < 0.001) a non-selective release of LH and FSH. In Experiments 2-4, Latin Square designs were utilized in vivo to examine whether physiological and hormonal milieu regulate putative selective effects of lGnRH-III. In Experiments 2 and 3, ovariectomized cows with basal levels of estradiol only (Experiment 2) or in combination with luteal phase levels of progester-one (Experiment 3) were injected with mGnRH and lGnRH-III (0.055, 0.11, 0.165 and 1.1 microg/kg body weight (BW) and saline. All doses of mGnRH released (P < 0.001) LH and FSH, but only the highest dose of lGnRH-III stimulated (P < 0.001) a non-selective release of both LH and FSH (Experiment 3). For Experiments 4A and 4B, intact, mid-luteal phase cows were injected with mGnRH and lGnRH-III (1.1 microg/kg BW; Experiment 4A), lGnRH-III (1.1 and 4.4 microg/kg BW; Experiment 4B) and saline. As before, mGnRH released (P < 0.001) both LH and FSH at all doses. In contrast, lGnRH-III at the highest dose released (P < 0.001) LH but not FSH. These findings suggest that lGnRH-III may act as a weak competitor for the mGnRH receptor and do not support the hypothesis that it selectively releases FSH in cattle.     

Relationships between energy balance and health traits of dairy cattle in early lactation

The objective of the study was to calculate phenotypic relationships between energy balance in early lactation and health and reproduction in that lactation. Data were 26,701 daily records of dry matter intake and milk production, periodic measures of milk composition and body weight, and all health and reproductive information from 140 multiparous Holstein cows. Daily energy balance was calculated by multiplying feed intake by the concentration of energy of the ration and subtracting the amount of energy required for maintenance (based on parity and body weight) and for milk production (based on yield and concentrations of fat, protein, and lactose). Six measures of energy balance were defined: mean daily energy balance during the first 20, 50, and 100 d of lactation; minimum daily energy balance; days in negative energy balance; and total energy deficit. Measures of health were the numbers of occurrences of each of the following during lactation: all udder problems, mastitis, all locomotive problems, laminitis, digestive problems, and reproductive problems. Reproductive traits were the number of days to first observed estrus and number of inseminations. Several significant relationships between energy balance and health were observed. Increased digestive and locomotive problems were associated with longer and more extreme periods of negative energy balance.     

Mitochondrial protein gene expression and the oxidative phosphorylation pathway associated with feed efficiency and energy balance in dairy cattle

Feed efficiency and energy balance are important traits underpinning profitability and environmental sustainability in animal production. They are complex traits, and our understanding of their underlying biology is currently limited. One measure of feed efficiency is residual feed intake (RFI), which is the difference between actual and predicted intake. Variation in RFI among individuals is attributable to the metabolic efficiency of energy utilization. High RFI (H_RFI) animals require more energy per unit of weight gain or milk produced compared with low RFI (L_RFI) animals. Energy balance (EB) is a closely related trait calculated very similarly to RFI. Cellular energy metabolism in mitochondria involves mitochondrial protein (MiP) encoded by both nuclear (NuMiP) and mitochondrial (MtMiP) genomes. We hypothesized that MiP genes are differentially expressed (DE) between H_RFI and L_RFI animal groups and similarly between negative and positive EB groups. Our study aimed to characterize MiP gene expression in white blood cells of H_RFI and L_RFI cows using RNA sequencing to identify genes and biological pathways associated with feed efficiency in dairy cattle. We used the top and bottom 14 cows ranked for RFI and EB out of 109 animals as H_RFI and L_RFI, and positive and negative EB groups, respectively. The gene expression counts across all nuclear and mitochondrial genes for animals in each group were used for differential gene expression analyses, weighted gene correlation network analysis, functional enrichment, and identification of hub genes. Out of 244 DE genes between RFI groups, 38 were MiP genes. The DE genes were enriched for the oxidative phosphorylation (OXPHOS) and ribosome pathways. The DE MiP genes were underexpressed in L_RFI (and negative EB) compared with the H_RFI (and positive EB) groups, suggestive of reduced mitochondrial activity in the L_RFI group. None of the MtMiP genes were among the DE MiP genes between the groups, which suggests a non-rate limiting role of MtMiP genes in feed efficiency and warrants further investigation. The role of MiP, particularly the NuMiP and OXPHOS pathways in RFI, was also supported by our gene correlation network analysis and the hub gene identification. We validated the findings in an independent data set. Overall, our study suggested that differences in feed efficiency in dairy cows may be linked to differences in cellular energy demand. This study broadens our knowledge of the biology of feed efficiency in dairy cattle.     

Group housing of Holstein calves in a poor indoor environment increases respiratory disease but does not influence performance or leukocyte responses

The objective of the current study was to determine if group-housing Holstein heifer calves in indoor pens with poor ventilation and drainage influences performance, health, leukocytes, and behavioral responses compared with individually housed calves. Ninety colostrum-fed calves (2 ± 1 d of age) were randomly assigned to 3 treatments: individually housed (G1; n = 30 calves), 2 calves per pen (G2; n = 30 calves), or 3 calves per pen (G3; n = 30 calves). The space allowance per calf was 2.5 m² for all treatment groups. All calves were fed 747 and 1,010 g/d of dry matter of a 28% CP and 20% fat milk replacer during the first 2 wk and wk 3 to 6, respectively. Weaning was initiated on d 46 by removing the evening feeding, and calves were completely weaned when they consumed 800 g/d of dry matter calf starter for 2 consecutive days after d 54. Calves were randomly commingled at d 90 in groups of 5 calves per pen in outdoors pens with an attached hutch. Peripheral blood was collected during the neonatal (3, 10, 21 d), weaning (46, 48, 54 d), and commingling periods (90, 93, 98 d) and was analyzed for neutrophil oxidative burst capacity when cocultured with an Escherichia coli, neutrophil surface L-selectin protein concentration, and whole blood secretion of tumor necrosis factor-α when cocultured with lipopolysaccharide. Behavior of each calf was assessed using 2 independent tests: an approach response to a human subject and the response of the calf when placed in an isolated cage. Calf starter intake was greater for G2 and G3 during wk 8 and 9, and also at wk 11 for G3 compared with G1. No treatment × time interaction or treatment effect for average daily gain was observed. Additionally, no treatment × time or treatment effect was noted for any leukocyte or biochemical variable of biological significance throughout the entire study. Individually housed calves tended to have a reduced incidence of respiratory disease during the first 90 d of life. No other treatment differences for the other health outcomes were observed. Group-housed calves also relied more on calf-to-calf interactions than calf-to-human interactions, as evident by their reduced approach of the human observer and more frequent movement when placed in an isolated pen. Group housing in an environment with poor ventilation and drainage may increase the risk of respiratory disease.     

Influence of concentrate allocation and energy balance on postpartum ovarian activity in Norwegian cattle

The effects of different levels of concentrate, milk yield, and energy balance (EB) on postpartum ovarian activity were studied in 164 lactations from 108 moderate yielding, dual-purpose cows. All cows were of the breed Norwegian Cattle. The cows were assigned to three different concentrate allocations and had free access to roughage. Cows in the high energy group were fed as suggested by Norwegian standards, whereas cows in the low energy group were supplied with approximately 40% less concentrate than recommended. Cows in the medium energy group were fed as the low energy group prepartum and as the high energy group during the first 90 d in milk (DIM). Energy balance was estimated by subtracting energy required for maintenance and lactation from energy intake. Ovarian activity was expressed in categories (early, mid and late responders) of days to first luteal response (milk progesterone concentration >5 ng/ml) and days to first regular cycle. Energy balance and milk yield throughout the first 100 DIM were compared between categories of concentrate allocation and ovarian activity using repeated measures mixed models. Decreasing level of concentrate was related to negative EB and declining milk yield. Both negative EB and high milk yield were associated with delayed postpartum ovarian activity. Concentrate allocation did not, however, alter days to first luteal response or days to first cycle. This study demonstrated that although negative EB was associated with delayed ovulation, Norwegian Cattle adapted to less energy-dense rations by reducing milk yield rather than delaying onset of ovarian activity.     

Effects of excess degradable protein on postpartum reproduction and energy balance in dairy cattle

The objectives of this study were to observe endocrine and reproductive responses of cows and heifers fed two diets (16 and 19% CP), which met undegradable protein requirements but differed in rumen degradable protein. Cows (n = 33) and heifers (n = 32) were randomly assigned within parity to diets at calving and remained on diets for 20 d after first breeding. Energy balance was determined twice weekly through the first luteal phase. Blood and milk samples were taken three times per week. Diet did not affect average daily energy balance, days to negative energy balance nadir, days to first ovulation, days to first service, or plasma glucose concentrations. First service conception rate was lower (31% vs. 48%) and plasma urea higher in animals fed the high protein diet. Days to energy balance nadir was correlated with days (r = .75) to first ovulation. Luteinizing hormone pulse frequency increased and pulse amplitude decreased in frequent samples (12-min intervals for 8 h) collected at 14 d postpartum versus sampling after the energy balance nadir. These data suggest that energy balance status plays an important role in determining the postpartum return of cyclic ovarian activity. Feeding excess CP as rumen degradable protein elevated plasma urea concentrations and decreased first service conception rate.     

Effects of brown midrib corn silage on the energy balance of dairy cattle

Effects of genotype and level of intake on net energy for lactation values of corn silage were evaluated by indirect calorimetry in two experiments using lactating and dry, nonpregnant dairy cows. In experiment 1, six multiparous Holstein cows in early lactation were fed experimental diets containing either brown midrib (bm3) or isogenic normal corn silage. Dietary treatments were isogenic and bm3 diets fed ad libitum, and the bm3 diets restricted-fed. Dry matter (DM) intake was 2.4 kg/d greater for cows fed the bm3 diet ad libitum compared with cows fed the isogenic diet. Apparent digestibilities of DM, organic matter, neutral detergent fiber, and acid detergent fiber were greater for cows restricted-fed bm3 than the isogenic diet. In experiment 2, six dry, nonpregnant Holstein cows were fed maintenance diets containing either bm3 or isogenic corn silage. Apparent digestibilities of DM, organic matter, neutral detergent fiber, and acid detergent fiber were greater for cows fed bm3 compared with isogenic corn silage. Digestible energy and metabolizable energy were greater for maintenance diets containing bm3 compared with isogenic corn silage, respectively. These data indicate increased milk production seen in other studies is a result of increased DMI rather than an increase in energy efficiency. Increased organic matter digestibility of bm3 corn silage resulted in greater digestible energy and metabolizable energy values in cows fed at maintenance energy intake. However, calculated net energy for lactation values of bm3 and isogenic corn silages were similar at both productive and maintenance levels of feeding.     

Genetic parameters for energy balance, feed efficiency, and related traits in Holstein cattle

Objectives of the current study were to estimate genetic parameters in Holstein cows for energy balance (EB) and related traits including dry matter intake (DMI), body weight (BW), body condition score (BCS), energy-corrected milk (ECM) production, and gross feed efficiency (GFE), defined as the ratio of total ECM yield to total DMI over the first 150d of lactation. Data were recorded for the first half of lactation on 227 and 175 cows in their first or later lactation, respectively. Random regression models were fitted to longitudinal data. Also, each trait was averaged over monthly intervals and analyzed by single and multivariate animal models. Heritability estimates ranged from 0.27 to 0.63, 0.12 to 0.62, 0.12 to 0.49, 0.63 to 0.72, and 0.49 to 0.53 for DMI, ECM yield, EB, BW, and BCS, respectively, averaged over monthly intervals. Daily heritability estimates ranged from 0.18 to 0.30, 0.10 to 0.26, 0.07 to 0.22, 0.43 to 0.67, and 0.25 to 0.38 for DMI, ECM yield, EB, BW, and BCS, respectively. Estimated heritability for GFE was 0.32. The genetic correlation of EB at 10d in milk (DIM) with EB at 150 DIM was -0.19, suggesting the genetic regulation of this trait differs by stage of lactation. Positive genetic correlations were found among DMI, ECM yield, and BW averaged over monthly intervals, whereas correlations of these traits with BCS depended upon stage of lactation. Total ECM yield for the lactation was positively correlated with DMI, but a negative genetic correlation between total ECM yield and EB was found. However, the genetic correlation between total ECM yield and EB in the first month of lactation was -0.02, indicating that total production is not genetically correlated with EB during the first month of lactation, when negative EB is most closely associated with diminished fitness. The genetic correlation between GFE and EB ranged from -0.73 to -0.99, indicating that selection for more efficient cows would favor a lower energy status. However, the genetic correlation between EB in the first month of lactation and GFE calculated from 75 to 150 DIM was not significant, indicating that the unfavorable correlation between GFE and EB in early lactation may be minimized with alternative definitions of efficiency. Thus, EB, GFE and related traits will likely respond to genetic selection in Holstein cows. However, the impact of selection for improved feed efficiency on EB must be carefully considered to avoid potential negative consequences of further reductions in EB at the onset of lactation.     

Expression of cell adhesion molecules, chemokines and chemokine receptors involved in leukocyte traffic in rats undergoing autoimmune orchitis

The testis is considered an immunologically privileged site where germ cell antigens are protected from autoimmune attack. Yet in response to infections, inflammatory diseases, or trauma, there is an influx of leukocytes to testicular interstitium. Interactions between endothelial cells (EC) and circulating leukocytes are implicated in the initiation and evolution of inflammatory processes. Chemokines are a family of chemoattractant cytokines characterized by their ability to both recruit and activate cells. Thus, we investigated the expression of CCL3, its receptors, and adhesion molecules CD31 and CD106 in an in vivo model of experimental autoimmune orchitis (EAO). In EAO, the highest content of CCL3 in testicular fluid coincides with onset of the disease. However, CCL3 released in vitro by testicular macrophages is higher during the immunization period. The specific chemokine receptors, CCR1 and CCR5, were expressed by testicular monocytes/macrophages and an increased number of CCR5+ cells was associated with the degree of testicular lesion. EC also play an essential role by facilitating leukocyte recruitment via their ability to express cell surface adhesion molecules that mediate interactions with leukocytes in the bloodstream. Rats with EAO showed a significant increase in the percentage of CD31+ EC that upregulate the expression of CD106. The percentage of leukocytes isolated from peripheral blood and lymph nodes expressing CD49d (CD106 ligand) also increases during orchitis. These data suggest that cell adhesion molecules, in conjunction with chemokines, contribute to the formation of a chemotactic gradient within the testis, causing the leukocyte infiltration characteristic of EAO histopathology.     

Tolerance does not develop to the decrease in nicotine self-administration produced by repeated bupropion administration.

The atypical antidepressant bupropion has been shown to be an efficacious smoking cessation agent; however, its therapeutic mechanism of action is unknown. To further understand the mechanism by which bupropion reduces smoking, the present study determined the effect of repeated bupropion pretreatment on nicotine self-administration or sucrose-maintained responding. Rats were trained to self-administer intravenous nicotine (0.02 mg/kg/infusion; Experiment 1) or to respond for sucrose pellets (45 mg each; Experiment 2) on a fixed-ratio 5 schedule. Once rats reached stable responding, bupropion (70 mg/kg, subcutaneously) or vehicle was injected 15 min before the session for 14 consecutive sessions. Bupropion acutely decreased both nicotine self-administration and sucrose-maintained responding by approximately 60%-70%. With repeated bupropion pretreatment, however, responding for nicotine decreased completely. In contrast, the bupropion-induced decrease in responding for sucrose following acute administration did not change significantly with repeated bupropion administration. These results suggest that bupropion acquired some specificity with repeated use, decreasing the intake of nicotine and producing an extinction-like pattern in nicotine self-administration. Thus the present results parallel human clinical studies with bupropion demonstrating its smoking cessation properties following repeated treatment. These results indicate that the rat nicotine self-administration paradigm is a useful animal model for assessing smoking cessation pharmacotherapies.     

Deletion of arginine from an abomasal infusion of amino acids does not decrease milk protein yield in Holstein cows

This study was undertaken to determine if a limited supply of Arg would alter milk and milk protein yields, as well as mammary uptake of AA and energetic substrates. Six lactating Holstein cows (199 ± 5 d in milk) were used in a replicated 3 × 3 Latin square balanced for residual effects with 14-d periods. The diet was formulated to supply 100% of the National Research Council net energy requirement and 72% of the metabolizable protein requirement. The treatments were randomly distributed as abomasal infusions of (1) water (CTL), (2) a mixture of essential AA (EAA) excluding Arg (ARG−), or (3) a mixture of EAA including Arg (ARG+). The profile of EAA in the infusates was the same as that found in casein with the exception that methionine was increased to maintain a 3:1 ratio of digestible lysine:methionine (total dietary + infusion). Milk protein yield was increased by the ARG+ compared with the CTL treatment and deletion of Arg in the infusate (ARG−) did not impair this response. Deletion of Arg from the EAA mixture decreased the mammary uptake of Arg relative to that of the CTL treatment, and although the uptake:output ratio decreased from 2.52 (ARG+) to 2.12 (ARG−), it was still largely in excess of Arg secretion in milk protein. Otherwise, Arg deletion did not affect any of the measured parameters (no significant difference between ARG− and ARG+) except Arg and urea arterial concentrations. In support of the increased yields of milk protein and lactose, mammary uptake of the group 2 AA (Ile, Leu, Lys, and Val) increased and the uptake:output ratio tended to increase from 1.04 to 1.23. The mammary uptake:milk protein output ratio was not different from 1 and not different among treatments for the group 1 AA (His, Met, Phe + Tyr, Trp). Mammary uptake of energetic substrates did not vary across treatments, although milk lactose yield increased with the ARG+ treatment relative to CTL. These results indicate that deletion of Arg has minimal effects on milk and milk component yields when the remaining EAA are supplied in sufficient amounts despite decreased mammary Arg uptake and that group 2 AA seem to be involved in the mammary gland to support the lactose yield response to EAA infusion.     

Reducing methane production with corn oil and calcium sulfate: Responses on whole-animal energy and nitrogen balance in dairy cattle

The addition of fat and calcium sulfate to diets fed to ruminants has resulted in a reduction in methane production, but the effects on energy balance have not been studied. A study using indirect calorimetry and 16 multiparous (8 Holstein and 8 Jersey; 78 ± 15 d in milk; mean ± standard deviation) lactating dairy cows was conducted to determine how mitigating methane production by adding corn oil or calcium sulfate to diets containing reduced-fat distillers grains affects energy and nitrogen balance. A replicated 4 × 4 Latin square design with 35-d periods (28 d of adaption and 4 d of collections) was used to compare 4 different dietary treatments. Treatments were composed of a control (CON) diet, which did not contain reduced-fat distillers grain and solubles (DDGS), and treatment diets containing 20% (dry matter basis) DDGS (DG), 20% DDGS with 1.38% (dry matter basis) added corn oil (CO), and 20% DDGS with 0.93% (dry matter basis) added calcium sulfate (CaS). Compared with CON, dry matter intake was not affected by treatment, averaging 29.6 ± 0.67 kg/d. Milk production was increased for diets containing DDGS compared with CON (26.3 vs. 27.8 ± 0.47 kg/d for CON vs. DDGS, respectively), likely supported by increased energy intake. Compared with CON, energy-corrected milk was greater in DG and CO (30.1 vs. 31.4, 31.7, and 31.0 ± 0.67 kg/d for CON, DG, CO, and CaS, respectively). Compared with CON, the addition of calcium sulfate and corn oil to diets containing DDGS reduced methane production per kg of dry matter intake (22.3, 19.9, and 19.6 ± 0.75 L/kg per d for CON, CO, and CaS, respectively). Similarly, methane production per kilogram of energy-corrected milk was reduced with the addition of calcium sulfate and corn oil to diets containing DDGS (14.2, 12.5, and 12.4 ± 0.50 L/kg per d for CON, CO, and CaS, respectively). Compared with CON and CaS, the intake of digestible energy was greater for DG and CO treatments (57.7, 62.1, 62.0, and 59.0 ± 1.38 Mcal/d for CON, DG, CO, and CaS, respectively). Intake of metabolizable energy was greater in all treatments containing DDGS compared with CON (50.5 vs. 54.0 ± 1.08 Mcal/d for CON vs. DDGS, respectively). Net balance (milk plus tissue energy) per unit of dry matter was greater in CO (containing DDGS and oil) than CON (1.55 vs. 1.35 ± 0.06 Mcal/kg for CO vs. CON, respectively). Tissue energy was greater in DG and CO compared with CON (6.08, 7.04, and 3.16 ± 0.99 Mcal/d for DG, CO, and CON, respectively. Results of this study suggest that the addition of oil and calcium sulfate to diets containing DDGS may be a viable option to reduce methane production and in the case of oil also improve net energy balance in lactating dairy cows.     

Overfeeding a moderate energy diet prepartum does not impair bovine subcutaneous adipose tissue insulin signal transduction and induces marked changes in peripartal gene network expression

Mechanisms regulating subcutaneous adipose tissue (SAT) insulin sensitivity and gene network expression during the peripartal period were evaluated in cows fed to meet or exceed prepartal energy requirements. Holstein cows were dried off at -50 d relative to expected parturition and fed a controlled-energy diet [CON; net energy for lactation=1.24 Mcal/kg of dry matter (DM); 36% of DM as wheat straw] until -21 d. Cows were then randomly assigned (n=7/diet) to either the same CON diet or a moderate-energy close-up diet (OVE; net energy for lactation=1.47 Mcal/kg of DM) until parturition. Biopsies of SAT were harvested at -10, 7, and 21 d for mRNA expression of 48 genes associated with insulin signaling, adipogenesis, and lipolysis. In vitro basal and insulin-stimulated insulin receptor substrate 1 tyrosine phosphorylation (IRS1-PY) was assessed at -10 and 7 d. The OVE led to more positive energy balance and greater serum insulin concentration prepartum. Compared with CON, OVE led to a more drastic increase in serum NEFA and also greater overall serum BHBA postcalving, both of which were associated with greater hepatic total lipid and triacylglycerol concentration. Close-up OVE did not improve any aspect of performance. In prepartal SAT, insulin-stimulated IRS1-PY was greater in OVE than in CON. However, IRS1-PY, serum insulin, and GLUT4 expression decreased postpartum regardless of prepartal treatment, suggesting a more severe state of insulin resistance. The expression of all genes encoding adipogenic regulators (PPARG and ZFP423), most lipogenic enzymes/inducers (FASN, SCD, DGAT2, and INSIG1), and basal-lipolysis regulators (ATGL and ABDH5) was greater at -10 d in OVE than in CON. Whereas adipogenic and basal lipolysis regulator expression remained greater in cows fed OVE by 7 d postpartum, expression of all lipogenic enzymes decreased regardless of diet. Despite those responses, the approximately 3-fold increase in expression of IRS1 and ZFP423 between 7 and 21 d suggested that insulin responsiveness and adipogenic capacity of SAT were partially restored. Expression of the preadipocyte marker DLK1, adiponutrin (PNPLA3), and fibroblast growth factor 21 (FGF21) was undetectable. Results suggested that close-up energy overfeeding did not exacerbate insulin resistance in SAT. Signs of restored insulin responsiveness (upregulation of IRS1, INSIG2, SREBF1, and ZFP423) were apparent as early as 3 wk postpartum. Thus, identifying specific nutrients capable of activating PPARγ after calving in AT might help accelerate its replenishment. A regulatory network encompassing the genes and physiological measurements obtained is proposed.     

Natural-light labeling of tomatoes does not facilitate growth or penetration of Salmonella into the fruit

The survival-growth capacity of Salmonella populations on tomato epidermis labeled by a natural-light labeling system was investigated after persistent fears of such marks serving as possible entryways for the pathogenic organisms, alone and in the presence of Pectobacterium carotovorum subsp. carotovorum, a soft-rot organism. Different treatments involving natural-light labeling, fruit waxing, and a five-strain cocktail of Salmonella were applied to mature green tomato surfaces in different sequences prior to storage at 4, 12, or 25°C. Fruit was sampled every 3 days, and Salmonella was enumerated from all treatments and unlabeled fruit, which served as controls. There were no significant differences between treatments or between treatments and controls throughout. The results indicate that the cuticle and epidermal interruptions caused by natural-light labeling do not facilitate the penetration and colonization of the tomato pericarp. In a separate set of experiments, the capacity of Salmonella to penetrate tomato in the presence of a potential synergism with P. carotovorum subsp. carotovorum was investigated. The addition of P. carotovorum at higher, lower, or equal population densities to Salmonella did not significantly alter the behavior of Salmonella on tomatoes stored at 25°C, regardless of natural-light labeling. The inability of P. carotovorum and Salmonella to colonize natural-light-etched surfaces of tomato fruit indicates that the use of this technology does not adversely compromise the surface of tomatoes.     

Limit-feeding a high-energy diet to meet energy requirements in the dry period alters plasma metabolite concentrations but does not affect intake or milk production in early lactation

Limit-feeding dry cows a high-energy diet may enable adequate energy intake to be sustained as parturition approaches, thus reducing the extent of negative energy balance after parturition. Our objective was to evaluate the effect of dry period feeding strategy on plasma concentrations of hormones and metabolites that reflect energy status. Multiparous Holstein cows (n = 18) were dried off 45 d before expected parturition, paired by expected calving date, parity, and previous lactation milk yield, and randomly assigned to 1 of 2 dry-period diets formulated to meet nutrient requirements at ad libitum or limited intakes. All cows were fed the same diet for ad libitum intake after parturition. Prepartum dry matter intake (DMI) for limit-fed cows was 9.4 kg/d vs. 13.7 kg/d for cows fed ad libitum. During the dry period, limit-fed cows consumed enough feed to meet calculated energy requirements, and ad libitum-fed cows were in positive calculated net energy for lactation (NE_L) balance (0.02 vs. 6.37 Mcal/d, respectively). After parturition, milk yield, milk protein concentration, DMI, body condition score, and body weight were not affected by the prepartum treatments. Cows limit fed during the dry period had a less-negative calculated energy balance during wk 1 postpartum. Milk fat concentration and yield were greater for the ad libitum treatment during wk 1 but were lower in wk 2 and 3 postpartum. Plasma insulin and glucose concentrations decreased after calving. Plasma insulin concentration was greater in ad libitum-fed cows on d −2 relative to calving, but did not differ by dietary treatment at other times. Plasma glucose concentrations were lower before and after parturition for cows limit-fed during the dry period. Plasma nonesterified fatty acid concentrations peaked after parturition on d 1 and 4 for the limit-fed and ad libitum treatments, respectively, and were greater for limit-fed cows on d −18, −9, −5, and −2. Plasma tumor necrosis factor-α concentrations did not differ by treatment in either the pre- or postpartum period, but tended to decrease after parturition. Apart from a reduction in body energy loss in the first week after calving, limit feeding a higher NE_L diet during the dry period had little effect on intake and milk production during the first month of lactation.     

2-Dodecylcyclobutanone does not induce mutations in the Salmonella mutagenicity test or intrachromosomal recombination in Saccharomyces cerevisiae

Treatment of foods, such as red meat and poultry, that contain palmitic acid with ionizing radiation leads to the formation of 2-dodecylcyclobutanone (2-DCB), a compound found only in irradiated foods. In this study, the Salmonella mutagenicity test and the yeast DEL assay were used to evaluate the genotoxic potential of 2-DCB. Salmonella Typhimurium tester strains TA98, TA100, TA1535, and TA1537 were exposed to 0, 0.125, 0.25, 0.5, and 1 mg per well of 2-DCB, with and without exogenous metabolic activation (5% S9 fraction), using the microtiter plate-based Miniscreen version of the test. 2-DCB did not induce mutations in the Salmonella mutagenicity test. When Saccharomyces cerevisiae strain RS112, which contains a nonfunctional duplication of the his3 gene that can be induced to form a functional HIS3+ gene by intrachromosomal recombination, was exposed to 0.63, 1.25, 2.5, or 5.0 mg/ml of 2-DCB, no increase in the rate of intrachromosomal (DEL) recombination was observed. The absence of genotoxicity observed in this study using purified 2-DCB agrees with the lack of genotoxic and teratogenic activity observed in previously conducted multigeneration feeding studies of laboratory animals (rats, mice, guinea pigs, and rabbits) that used radiation-sterilized poultry that contained 2-DCB as a unique radiolytic product.     

Chicory inulin does not increase stool weight or speed up intestinal transit time in healthy male subjects

Inulin is a non-digestible oligosaccharide classified as a prebiotic, a substrate that promotes the growth of certain beneficial microorganisms in the gut. We examined the effect of a 20 g day(-1) supplement of chicory inulin on stool weight, intestinal transit time, stool frequency and consistency, selected intestinal microorganisms and enzymes, fecal pH, short chain fatty acids and ammonia produced as by-products of bacterial fermentation. Twelve healthy male volunteers consumed a well-defined, controlled diet with and without a 20 g day(-1) supplement of chicory inulin (degree of polymerization (DP) ranging for 2-60), with each treatment lasting for 3 weeks in a randomized, double-blind crossover trial. Inulin was consumed in a low fat ice cream. No differences were found in flavor or appeal between the control and inulin-containing ice creams. Inulin consumption resulted in a significant increase in total anaerobes and Lactobacillus species and a significant decrease in ammonia levels and β-glucuronidase activity. Flatulence increased significantly with the inulin treatment. No other significant differences were found in bowel function with the addition of inulin to the diet. Thus, inulin is easily incorporated into a food product and has no negative effects on food acceptability. Twenty grams of inulin was well tolerated, but had minimal effects on measures of laxation in healthy, human subjects.