Candice M. Klingerman

The effect of Lactobacillus buchneri 40788 or Lactobacillus plantarum MTD-1 on the fermentation and aerobic stability of corn silages ensiled at two dry matter contents

Whole-plant corn was harvested at 33 (normal) and 41% (moderately high) dry matter (DM) and ensiled in quadruplicate 20-L laboratory silos to investigate the effects of Lactobacillus buchneri 40788 (LB) or L. plantarum MTD-1 (LP) alone, or in combination, on the fermentation and aerobic stability of the resulting silage. Aerobic stability was defined as the amount of time after exposure to air for the silage temperature to reach 2 degrees C above ambient temperature. The chopped forage was used in a 2 x 2 x 2 factorial arrangement of treatments: normal and moderately high DM contents, LB at 0 (untreated) or 4 x 10(5) cfu/g of fresh forage, and LP at 0 or 1 x 10(5) cfu/g. After 240 d of ensiling, corn silage harvested at the moderately high DM had higher pH, higher concentrations of ethanol, and more yeasts compared with the silage ensiled at the normal DM content. Inoculation with LB did not affect the concentration of lactic acid in silages with a moderately high DM, but decreased the concentration of lactic acid in the silage with normal DM. Higher concentrations of acetic acid were found in the silage treated with LB compared with those not treated with this organism. Inoculation with LP increased the concentration of lactic acid only in the silage with the normal DM content. The concentration of acetic acid was lower in silage treated with LP with a moderately high DM content, but greater in the silage treated with LP with the normal DM content when compared with silages without this inoculant. Appreciable amounts of 1,2-propanediol (average 1.65%, DM basis) were found in all silages treated with LB regardless of the DM content. The addition of L. buchneri increased the concentration of NH(3)-N in silages but the addition of L. plantarum decreased it. Aerobic stability was improved in all silages treated with LB, with greater aerobic stability occurring in the silage with moderately high DM compared with silage with normal DM content. Inoculation with LP had no effect on aerobic stability. There were no interactions between L. buchneri and L. plantarum for most fermentation products or aerobic stability of the silages. This study showed that inoculating whole-plant corn with L. buchneri 40788 or L. plantarum MTD-1 has different beneficial effects on the resulting silage. There appear to be no major interactions between these organisms when added together to forage. Thus, there is potential to add both organisms simultaneously to improve the fermentation and aerobic stability of corn silage.

Food Restriction-Induced Changes in Gonadotropin-Inhibiting Hormone Cells are Associated with Changes in Sexual Motivation and Food Hoarding, but not Sexual Performance and Food Intake

We hypothesized that putative anorectic and orexigenic peptides control the motivation to engage in either ingestive or sex behaviors, and these peptides function to optimize reproductive success in environments where energy fluctuates. Here, the putative orexigenic peptide, gonadotropin-inhibiting hormone (GnIH, also known as RFamide-related peptide-3), and the putative anorectic hormones leptin, insulin, and estradiol were examined during the course of food restriction. Groups of female Syrian hamsters were restricted to 75% of their ad libitum food intake or fed ad libitum for 4, 8, or 12 days. Two other groups were food-restricted for 12 days and then re-fed ad libitum for 4 or 8 days. After testing for sex and ingestive behavior, blood was sampled and assayed for peripheral hormones. Brains were immunohistochemically double-labeled for GnIH and the protein product of the immediate early gene, c-fos, a marker of cellular activation. Food hoarding, the number of double-labeled cells, and the percent of GnIH-Ir cells labeled with Fos-Ir were significantly increased at 8 and 12 days after the start of food restriction. Vaginal scent marking and GnIH-Ir cell number significantly decreased after the same duration of restriction. Food hoarding, but not food intake, was significantly positively correlated with cellular activation in GnIH-Ir cells. Vaginal scent marking was significantly negatively correlated with cellular activation in GnIH-Ir cells. There were no significant effects of food restriction on plasma insulin, leptin, estradiol, or progesterone concentrations. In the dorsomedial hypothalamus (DMH) of energetically challenged females, strong projections from NPY-Ir cells were found in close apposition to GnIH-Ir cells. Together these results are consistent with the idea that metabolic signals influence sexual and ingestive motivation via NPY fibers that project to GnIH cells in the DMH.

An evaluation of exogenous enzymes with amylolytic activity for dairy cows.

An experimental (7B) and a commercial (AMA) formulation of enzymes, both primarily with alpha-amylase activity, were evaluated for activity at various pH values, stability in ruminal fluid, the potential to improve in vitro ruminal fermentations, and the potential to improve production performance of lactating cows. When incubated (40 degrees C) in buffer with a pH between 5.4 and 6.0, 7B had about 10 to 25 times greater amylase activity than AMA, and enzyme activity in this range increased by 100% for 7B, whereas activity decreased by about 26% for AMA. Both formulations maintained enzyme activity when they were incubated in in vitro ruminal fermentations for 24 h. After 6 h of ruminal in vitro fermentation, additions of 7B resulted in linear increases in apparent total volatile fatty acid production for flint and dent corn but had no effect on floury corn. In a lactation trial, 28 Holstein cows (68 +/- 31 d in milk, 46.9 +/- 9.1 kg of milk/d) were fed a total mixed ration (TMR) supplemented with nothing (CON), a low dose of 7B [7BL, 0.88 mL/kg of TMR dry matter (DM)], a high dose of 7B (7BH, 4.4 mL/kg of TMR DM), or AMA (0.4 g/kg of TMR DM). The experiment was conducted as a 4. 4 Latin square design with 21-d periods. Cows fed 7BL, 7BH, and AMA ate similar amounts of DM, and cows fed the latter 2 diets consumed more DM than did cows fed CON. Cows fed 7BL produced more milk than cows fed CON and 7BH, but produced similar amounts to cows fed AMA. The production of 3.5% fat-corrected milk was greater from cows fed 7BL and AMA compared with cows fed CON. The percentages of milk fat and milk protein were unaffected by treatment. Total-tract digestion of DM and organic matter were greater for cows fed 7BL compared with those fed CON. The addition of exogenous amylase enzymes to the diets of lactating dairy cows has the potential to improve animal productivity.

H2S concentrations in the arterial blood during H2S administration in relation to its toxicity and effects on breathing

Our aim was to establish in spontaneously breathing urethane-anesthetized rats, the relationship between the concentrations of H2S transported in the blood and the corresponding clinical manifestations, i.e., breathing stimulation and inhibition, during and following infusion of NaHS at increasing rates. The gaseous concentration of H2S (CgH2S, one-third of the total soluble form) was computed from the continuous determination of H2S partial pressure in the alveolar gas, while H2S, both dissolved and combined to hemoglobin, was measured at specific time points by sulfide complexation with monobromobimane (CMBBH2S). We found that using a potent reducing agent in vitro, H2S added to the whole blood had little interaction with the plasma proteins, as sulfide appeared to be primarily combined and then oxidized by hemoglobin. In vivo, H2S was undetectable in the blood in its soluble form in baseline conditions, while CMBBH2S averaged 0.7 ± 0.5 μM. During NaHS infusion, H2S was primarily present in nonsoluble form in the arterial blood: CMBBH2S was about 50 times higher than CgH2S at the lowest levels of exposure and 5 or 6 times at the levels wherein fatal apnea occurred. CgH2S averaged only 1.1 ± 0.7 μM when breathing increased, corresponding to a CMBBH2S of 11.1 ± 5.4 μM. Apnea occurred at CgH2S above 5.1 μM and CMBBH2S above 25.4 μM. At the cessation of exposure, CMBBH2S remained elevated, at about 3 times above baseline for at least 15 min. These data provide a frame of reference for studying the putative effects of endogenous H2S and for testing antidotes against its deadly effects.

In-Vivo Interactions Between Cobalt or Ferric Compounds and the Pools of Sulphide in the Blood During and After H2s Poisoning

Hydrogen sulphide (H2S), a chemical hazard in oil and gas production, has recently become a dreadful method of suicide, posing specific risks and challenges for the first responders. Currently, there is no proven effective treatment against H2S poisoning and its severe neurological, respiratory or cardiac after-effects. We have recently described that H2S is present in various compartments, or pools, in the body during sulphide exposure, which have different levels of toxicity. The general goals of our study were to (1) determine the concentrations and kinetics of the various pools of hydrogen sulphide in the blood, i.e., gaseous (CgH2S) versus total sulphide, i.e., reacting with monobromobimane (CMBBH2S), during and following H2S exposure in a small and large mammal and (2) establish the interaction between the pools of H2S and a methemoglobin (MetHb) solution or a high dose of hydroxocobalamin (HyCo). We found that CgH2S during and following H2S infusion was similar in sedated sheep and rats at any given rate of infusion/kg and provoked symptoms, i.e., hyperpnea and apnea, at the same CgH2S. After H2S administration was stopped, CgH2S disappeared within 1 min. CMBBH2S also dropped to 2-3μM, but remained above baseline levels for at least 30 min. Infusion of a MetHb solution during H2S infusion produced an immediate reduction in the free/soluble pool of H2S only, whereas CMBBH2S increased by severalfold. HyCo (70 mg/kg) also decreased the concentrations of free/soluble H2S to almost zero; CgH2S returned to pre-HyCo levels within a maximum of 20 min, if H2S infusion is maintained. These results are discussed in the context of a relevant scenario, wherein antidotes can only be administered after H2S exposure.

Energetic challenges unmask the role of ovarian hormones in orchestrating ingestive and sex behaviors

Effects of ovarian hormones on sex and ingestive behavior are well studied, and yet, their role in diverting attention from food to sex has not been examined directly, possibly because these functions are masked under conditions of excessive food abundance typical of the laboratory. Female Syrian hamsters were either fed ad libitum or food-restricted to 75% of their ad libitum intake for 8days and then tested every day of the estrous cycle for their preference for males versus food, food hoarding and food intake in an apparatus designed to mimic aspects of their natural habitat. The food-restricted, but not the fed females, varied significantly over the estrous cycle in appetitive behaviors, which included their preference for males versus food and in the amount of food hoarded, with low food hoarding and high male preference on the night of ovulation. In contrast, there were no significant differences between restricted and ad libitum-fed females in the consummatory behaviors, namely, food intake or lordosis duration. In ovariectomized females, estradiol plus progesterone treatment delayed food restriction-stimulated hoarding and hastened feeding-inhibited hoarding without affecting food intake or lordosis duration. In summary, energy restriction and the presence of males unmasked an effect that was obscured in the normal laboratory conditions characterized by isolation and an over abundance of readily available food. These results are consistent with the idea that ovarian hormones orchestrate appetites for food and sex to optimize reproductive success under fluctuating energetic conditions.

Sense and nonsense in metabolic control of reproduction.

An exciting synergistic interaction occurs among researchers working at the interface of reproductive biology and energy homeostasis. Reproductive biologists benefit from the theories, experimental designs, and methodologies used by experts on energy homeostasis while they bring context and meaning to the study of energy homeostasis. There is a growing recognition that identification of candidate genes for obesity is little more than meaningless reductionism unless those genes and their expression are placed in a developmental, environmental, and evolutionary context. Reproductive biology provides this context because metabolic energy is the most important factor that controls reproductive success and gonadal hormones affect energy intake, storage, and expenditure. Reproductive hormone secretion changes during development, and reproductive success is key to evolutionary adaptation, the process that most likely molded the mechanisms that control energy balance. It is likely that by viewing energy intake, storage, and expenditure in the context of reproductive success, we will gain insight into human obesity, eating disorders, diabetes, and other pathologies related to fuel homeostasis. This review emphasizes the metabolic hypothesis: a sensory system monitors the availability of oxidizable metabolic fuels and orchestrates behavioral motivation to optimize reproductive success in environments where energy availability fluctuates or is unpredictable.

Food restriction dissociates sexual motivation, sexual performance, and the rewarding consequences of copulation in female Syrian hamsters.

Animals can switch their behavioral priorities from ingestive to sex behaviors to optimize reproductive success in environments where energy fluctuates. We hypothesized that energy availability differentially affects the appetitive (motivation), consummatory (performance), and learned (rewarding) components of behavior. In Experiment 1, appetitive and consummatory aspects of sex behavior were dissociated in the majority of female Syrian hamsters restricted to 75% of their ad libitum food intake for between 8 and 11 days. Food restriction significantly inhibited vaginal scent marking, decreased the preference for spending time with male hamsters vs. spending time with food, and increased food hoarding with no significant effect on consummatory behaviors such as the incidence of lordosis or food intake. In Experiments 2 and 3, we attempted to use a similar level of food restriction to dissociate sexual appetite from sexual reward. In hamsters, formation of a conditioned place preference (CPP) for copulatory reward is reflected in increased nucleus accumbens (NAc) neural activation, measured as immunocytochemical staining for c-Fos, the protein product of the immediate-early gene, c-fos. In Experiment 2, neural activation increased 1h after copulation in the NAc, and did not differ significantly between 10-day food-restricted and ad libitum-fed females in any brain area examined. In Experiment 3, females were either food-restricted or fed ad libitum over 8-30 days of conditioning with copulatory stimuli. Food-restricted females showed significantly fewer appetitive behaviors, but no difference in formation of a CPP compared to females fed ad libitum. Together these data are consistent with the idea that mild levels of food restriction that inhibit appetitive behaviors fail to attenuate consummatory behaviors and the rewarding consequences of copulation. Thus, appetitive sex behaviors are, at least partially, neuroanatomically and behaviorally distinct from both consummatory behaviors and copulatory reward.

The effect of wide swathing on wilting times and nutritive value of alfalfa haylage.

On 3 consecutive cuttings, alfalfa from a single field was mowed with a John Deere 946 mower-conditioner (4-m cut width; Moline, IL) to leave narrow swaths (NS) ranging from 1.2 to 1.52 m wide (30-37% of cutter bar width) and wide swaths (WS) ranging from 2.44 to 2.74 m wide (62-67% of cutter bar width). Samples were collected from windrows and dry matter (DM) was monitored during wilting until a target of 43 to 45% DM was obtained. Forage from random windrows (n=4-6) was harvested by hand, chopped through a forage harvester before being packed in replicated vacuum-sealed bags, and allowed to ensile for 65 d. There was no swath width x cutting interaction for any parameter tested. Over all cuttings, the resulting silage DM was not different between the NS silage (43.8%) and the WS silage (44.9%). However, wide swathing greatly reduced the time of wilting before making silage. The hours of wilting time needed to reach the targeted DM for the NS silage compared with the WS silage at cuttings 1, 2, and 3 were 50 versus 29, 54 versus 28, and 25 versus 6, respectively. At the time of ensiling, the WS silage had more water-soluble carbohydrates (5.1%) than did the NS silage (3.7%). The WS silage had a lower pH (4.58) than did the NS silage (4.66), but swath width did not affect fermentation end products (lactic acid, acetic acid, and ethanol). The NS silage had more NH(3)-N (0.26%) than did the WS silage (0.21%). Wide swathing did not affect the concentration of ash or the digestibility of NDF, but it lowered the N content (NS=3.45%; WS=3.23%) and increased the ADF content (NS=39.7%; WS=40.9%) of the resulting silage. Wide swathing can markedly reduce the time that alfalfa must wilt before it can be chopped for silage, but under good conditions, as in this study, the resulting silage quality was generally not improved.

Second-Generation Antipsychotics Cause a Rapid Switch to Fat Oxidation That Is Required for Survival in C57BL/6J Mice

Some second-generation antipsychotics (SGAs) increase insulin resistance and fat oxidation, but counter intuitively they do not activate lipolysis. This seems unsustainable for meeting energy demands. Here, we measured dose-dependent effects of SGAs on rates of oxygen consumption (VO2), respiratory exchange ratio (RER), and physical activity in C57BL/6J mice. The role of H1-histamine receptors and consequences of blocking fat oxidation were also examined. Olanzapine, risperidone, and clozapine (2.5-10mg/kg) elicited rapid drops in dark-cycle RER (~0.7) within minutes, whereas aripiprazole exerted only modest changes. Higher doses of olanzapine decreased VO2, and this was associated with accumulation of glucose in plasma. Clozapine and risperidone also lowered VO2, in contrast to aripiprazole, whereas all decreased physical activity. Astemizole and terfenadine had no significant effects on RER, VO2, or physical activity. The VO2 and RER effects appear independent of sedation/physical activity or H1-receptors. CPT-1 inhibitors can enhance muscle glucose utilization and prevent fat oxidation. However, after etomoxir (2 × 30 mg/kg), a low dose of olanzapine that did not significantly affect VO2 by itself caused precipitous drops in VO2 and body temperature, leading to death within hours or a moribund state requiring euthanasia. One 30 mg/kg dose of either etomoxir or 2-tetradecylglycidate followed by olanzapine, risperidone, or clozapine, but not aripiprazole, dramatically lowered VO2 and body temperature. Thus, mice treated with some SGAs shift their fuel utilization to mostly fat but are unable to either switch back to glucose or meet their energy demands when either higher doses are used or when fat oxidation is blocked.