Coordinated changes in energy intake and expenditure following hypothalamic administration of neuropeptides involved in energy balance.

OBJECTIVE: The hypothalamic control of energy balance is regulated by a complex network of neuropeptide-releasing neurons. Although the effect of these neuropeptides on individual aspects of energy homoeostasis has been studied, the coordinated response of these effects has not been comprehensively investigated. We have simultaneously monitored a number of metabolic parameters following intracerebroventricular (ICV) administration of 1 and 3 nmol of neuropeptides with established roles in the regulation of feeding, activity and metabolism. Ad libitum- fed rats received the orexigenic neuropeptides neuropeptide Y (NPY), agouti-related protein (AgRP), melanin-concentrating hormone (MCH) or orexin-A. Overnight-food-deprived rats received an ICV injection of the anorectic peptides alpha-melanocyte-stimulating hormone (MSH), corticotrophin-releasing factor (CRF) or neuromedin U (NMU). RESULTS: Our results reveal the temporal sequence of the effects of these neuropeptides on both energy intake and expenditure, highlighting key differences in their function as mediators of energy balance. NPY and AgRP increased feeding and decreased oxygen consumption, with the effects of AgRP being more prolonged. In contrast, orexin-A increased both feeding and oxygen consumption, consistent with an observed increase in activity. The potent anorexigenic effects of CRF were accompanied by a prolonged increase in activity, whereas NMU injection resulted in significant but short-lasting inhibition of food intake, ambulatory activity and oxygen consumption. alpha-MSH injection resulted in significant increases in both ambulatory activity and oxygen consumption, and reduced food intake following administration of 3 nmol of the peptide. CONCLUSION: We have for the first time, simultaneously measured several metabolic parameters following hypothalamic administration of a number of neuropeptides within the same experimental system. This work has shown the interrelated effects of these neuropeotides on activity, energy expenditure and food intake, thus facilitating comparison between the different hypothalamic systems.

Metabolism of dairy cows as affected by prepartum dietary carbohydrate source and supplementation with chromium throughout the periparturient period.

Holstein cows (n = 72) entering second or later lactation were used to determine whether metabolic indices and hepatic capacities for oxidation and gluconeogenesis from propionate are affected by source of carbohydrate in the prepartum diet and chromium-l-methionine (Cr-Met) supplementation throughout the periparturient period. Cows were fed prepartum diets as total mixed rations with the concentrate portion based either on starch-based cereals [high nonfiber carbohydrate (NFC); 1.59 Mcal/kg of net energy for lactation (NE(L)), 14.4% crude protein (CP), 40.3% NFC] or nonforage fiber sources (low NFC; 1.54 Mcal/kg of NE(L), 14.5% CP, 33.6% NFC) from 21 d before expected parturition until parturition. After parturition all cows were fed a common lactation total mixed ration (1.74 Mcal/kg of NE(L), 16.5% CP, 40.0% NFC). The Cr-Met was supplemented once daily via gelatin capsule at dosages of 0, 0.03, or 0.06 mg of Cr/kg of BW(0.75). Thus, treatments were in a 2 (carbohydrate source) x 3 (Cr-Met) factorial arrangement. There was no effect of prepartum carbohydrate source on pre- and postpartum plasma concentrations of glucose, nonesterified fatty acids (NEFA), beta-hydroxybutyrate (BHBA), insulin, glucagon, or insulin to glucagon ratio. However, cows fed the low NFC diet during the prepartum period tended to have greater plasma NEFA and lower BHBA concentrations postpartum. Liver glycogen concentrations tended to be greater on d 1 postpartum for cows fed low NFC prepartum. Supplementing 0.03 mg/kg of BW(0.75) of Cr as Cr-Met increased prepartum plasma glucose and glucagon concentrations and tended to decrease prepartum plasma NEFA concentrations compared with either 0 or 0.06 mg of Cr/kg of BW(0.75). Postpartum plasma glucose concentrations decreased linearly and glucagon concentrations were increased quadratically by administering increasing amounts of Cr-Met. Supplementing Cr-Met did not affect prepartum plasma concentrations of insulin or BHBA, postpartum NEFA or BHBA, or liver composition. There was an interaction of prepartum carbohydrate source and Cr-Met supplementation such that in vitro hepatic conversion of [1-(14)C]propionate to both CO(2) and glucose was similar or increased when Cr-Met was supplemented to cows fed the low NFC diet but decreased when Cr-Met was supplemented to cows fed the high NFC diet. Insulin addition in vitro did not affect hepatic metabolism of propionate on d 1 postpartum. Overall, both the NFC content of the prepartum diet and Cr-Met had only modest effects on metabolic indices in this experiment.

Hypothalamic mapping of orexigenic action and Fos-like immunoreactivity following relaxin-3 administration in male Wistar rats.

The insulin superfamily, characterized by common disulphide bonds, includes not only insulin but also insulin-like peptides such as relaxin-1 and relaxin-3. The actions of relaxin-3 are largely unknown, but recent work suggests a role in regulation of food intake. Relaxin-3 mRNA is highly expressed in the nucleus incertus, which has extensive projections to the hypothalamus, and relaxin immunoreactivity is present in several hypothalamic nuclei. In the rat, relaxin-3 binds and activates both relaxin family peptide receptor 1, which also binds relaxin-1, and a previously orphaned G protein-coupled receptor, RXFP3. These receptors are extensively expressed in the hypothalamus. The aims of these studies were twofold: 1) map the hypothalamic site(s) of the orexigenic action of relaxin-3 and 2) examine the site(s) of neuronal activation following central relaxin-3 administration. After microinjection into hypothalamic sites, human relaxin-3 (H3; 180 pmol) significantly stimulated 0- to 1-h food intake in the supraoptic nucleus (SON), arcuate nucleus (ARC), and the anterior preoptic area (APOA) [SON 0.4+/-0.2 (vehicle) vs. 2.9+/-0.5 g (H3), P<0.001; ARC 0.7+/-0.3 (vehicle) vs. 2.7+/-0.2 g (H3), P<0.05; and APOA 0.8+/-0.1 (vehicle) vs. 2.2+/-0.2 g (H3), P<0.05]. Cumulative food intake was significantly increased

Microinjection of galanin-like peptide into the medial preoptic area stimulates food intake in adult male rats.

Galanin-like peptide (GALP) is a neuropeptide implicated in the regulation of feeding behaviour, metabolism and reproduction. GALP is an endogenous ligand of the galanin receptors, which are widely expressed in the hypothalamus. GALP is predominantly expressed in arcuate nucleus (ARC) neurones, which project to the paraventricular nucleus (PVN) and medial preoptic area (mPOA). Intracerebroventricular or intraparaventricular (iPVN) injection of GALP acutely increases food intake in rats. The effect of GALP injection into the mPOA on feeding behaviour has not previously been studied. In the present study, intra-mPOA (imPOA) injection of GALP potently increased 0-1-h food intake in rats. The dose-response effect of imPOA GALP administration on food intake was similar to that previously observed following iPVN administration. The effects of GALP (1 nmol) or galanin (1 nmol) on food intake were then compared following injection into the PVN, mPOA, ARC, dorsal medial nucleus (DMN), lateral hypothalamus and rostral preoptic area (rPOA). GALP (1 nmol) increased food intake to a similar degree when injected into the imPOA or iPVN, but produced no significant effect when injected into the ARC, DMN, lateral hypothalamus or rPOA. Similarly, galanin (1 nmol) significantly increased food intake following injection imPOA and iPVN. However, the effect was significantly smaller than that following administration of GALP (1 nmol). Galanin also had no significant effect on food intake when administered into the ARC, DMN, lateral hypothalamus and rPOA. These data suggest that the mPOA and the PVN may have specific roles in mediating the orexigenic effect of GALP and galanin.

Central relaxin-3 administration causes hyperphagia in male Wistar rats.

Relaxin-3 (INSL-7) is a recently discovered member of the insulin superfamily. Relaxin-3 mRNA is expressed in the nucleus incertus of the brainstem, which has projections to the hypothalamus. Relaxin-3 binds with high affinity to the LGR7 receptor and to the previously orphan G protein-coupled receptor GPCR135. GPCR135 mRNA is expressed predominantly in the central nervous system, particularly in the paraventricular nucleus (PVN). The presence of relaxin-3 and these receptors in the PVN led us to investigate the effect of central administration of relaxin-3 on food intake in male Wistar rats. The receptor involved in mediating these effects was also investigated. Intracerebroventricular injections of human relaxin-3 (H3) to satiated rats significantly increased food intake 1 h post administration in the early light phase [0.96 +/- 0.16 g (vehicle) vs. 1.81 +/- 0.21 g (180 pmol H3), P < 0.05] and the early dark phase [2.95 +/- 0.45 g (vehicle) vs. 4.39 +/- 0.39 g (180 pmol H3), P < 0.05]. Intra-PVN H3 administration significantly increased 1-h food intake in satiated rats in the early light phase [0.34 +/- 0.16 g (vehicle) vs. 1.23 +/- 0.30 g (18 pmol H3), P < 0.05] and the early dark phase [4.43 +/- 0.32 g (vehicle) vs. 6.57 +/- 0.42 g (18 pmol H3), P < 0.05]. Feeding behavior increased after intra-PVN H3. Equimolar doses of human relaxin-2, which binds the LGR7 receptor but not GPCR135, did not increase feeding. Hypothalamic neuropeptide Y, proopiomelanocortin, or agouti-related peptide mRNA expression did not change after acute intracerebroventricular H3. These results suggest a novel role for relaxin-3 in appetite regulation.

Performance of dairy cows as affected by prepartum dietary carbohydrate source and supplementation with chromium throughout the transition period.

Holstein cows (n = 72) entering second or later lactation were used to determine whether productive performance and dry matter intake (DMI) are affected by carbohydrate source in the prepartum diet and chromium-L-methionine (Cr-Met) supplementation throughout the periparturient period. Cows were fed either a TMR with the concentrate portion based on starch-based cereals [high nonfiber carbohydrate (NFC); 1.59 Mcal/kg of net energy for lactation (NEL), 14.4% crude protein (CP), 40.3% NFC] or a TMR with the concentrate portion based on nonforage fiber sources (low NFC; 1.54 Mcal/kg NEL, 14.5% CP, 33.6% NFC) from 21 d before expected parturition until parturition. After parturition all cows were fed a lactation TMR (1.74 Mcal/kg NEL, 16.5% CP, 40.0% NFC). The Cr-Met was supplemented once daily via gelatin capsule at dosages of 0, 0.03, or 0.06 mg of Cr/kg of metabolic body weight. Thus, treatments were in a 2 (carbohydrate source) x 3 (Cr-Met) factorial arrangement. Neither prepartum nor postpartum DMI was affected by prepartum dietary carbohydrate source. Administering increasing amounts of Cr-Met linearly increased milk yield and, subsequently, postpartum DMI. Prepartum carbohydrate source did not affect postpartum milk yield; however, cows fed the low NFC diet tended to yield milk with a lower content of total solids. These data indicate that prepartum carbohydrate source has little influence on performance during the immediate peripartal period, and that increases in milk yield for cows supplemented with Cr-Met are independent of prepartum dietary carbohydrate source.

Effect of immunoglobulin G from cows immunized with ferric citrate receptor (FecA) on iron uptake by Escherichia coli.

The effects of immunoglobulin (Ig) G from cows immunized with the ferric citrate receptor (FecA) on iron uptake by Escherichia coli were investigated. Receptor FecA was purified from E. coli UT5600/pSV66. Cows were immunized with 400 microg purified FecA three times at 21 d intervals during late lactation and the nonlactating period. Immunoglobulin G was purified by protein G affinity chromatography from colostral whey from cows immunized with FecA and from unimmunized control cows. The purified IgG from FecA immunized cows had higher IgG titers against FecA compared with control IgG. Fifteen E. coli isolated from intramammary infections and E. coli UT5600/pSV66 were grown in an iron-depleted medium containing 1 mM citrate to induce FecA. The bacterial cells were mixed with 0, 2, and 4 mg/ml purified IgG, and 55Fe was added to the assay. After 5, 10, and 15 min incubations at 37 degrees C, samples were passed through 0.45-pm pore size filters. Filters were washed with saline three times, and the radioactivity of 55Fe taken up by the bacterial cells on the filters was measured by a liquid scintillation counter. The measurements were expressed as numbers of 55Fe atoms per colony-forming unit and transformed to log10. The assay was repeated three times for each isolate in a partially balanced incomplete block design. The presence of IgG decreased 55Fe uptake by E. coli mastitis isolates and E. coli UT5600/pSV66. Anti-FecA IgG reduced 55Fe uptake by E. coli greater than IgG from unimmunized cows.

Growth responses of coliform bacteria to purified immunoglobulin G from cows immunized with ferric enterobactin receptor FepA.

The ability of purified bovine immunoglobulin (Ig) G from cows immunized with ferric enterobactin receptor FepA to inhibit the growth of coliform bacteria derived from bovine intramammary infection was investigated in iron-restricted media. All isolates of Escherichia coli (n = 21) and Klebsiella pneumoniae (n = 21) were tested for growth in a chemically defined medium containing 0.5 mg/ml of apolactoferrin and in a pooled source of dry cow secretion. The addition of 4 mg/ml of purified bovine IgG directed against FepA in the synthetic medium resulted in significant growth inhibition for both E. coli and K. pneumoniae isolates. Growth reduction of E. coli was greater than that of K. pneumoniae. In dry cow secretions, the growth of each E. coli isolate but of less than half of K. pneumoniae isolates (43%) was inhibited by IgG from cows immunized with FepA. Purified bovine IgG from cows immunized with E. coli J5 had a minimal inhibitory effect on the growth of both E. coli and K. pneumoniae isolates in the synthetic medium. In dry cow secretions, IgG from cows immunized with E. coli and K. pneumoniae isolates. Supplementation with 50 microM of ferric chloride to the medium completely reversed the inhibitory effects of the antibodies and lactoferrin. Bovine IgG directed against FepA apparently inhibited the growth of coliform bacteria by interfering with the binding of the ferric enterobactin complex to the cell surface receptor FepA.

Effect of vitamin E supplementation in diets with a low concentration of selenium on mammary gland health of dairy cows.

Sixty-six cows and heifers (Holsteins and Jerseys) were assigned to one of three treatments at 60 d before anticipated calving. Treatment 1 consisted of 100 IU/d of supplemental vitamin E during the dry period and 100 IU/d during the first 30 d of lactation. Treatment 2 was 1000 IU/d of vitamin E during the dry period and 500 IU/d during lactation. Treatment 3 was 1000 IU/d of vitamin E during the first 46 d of the dry period, 4000 IU/d during the last 14 d of the dry period, and 2000 IU/d during lactation. Plasma concentrations of alpha-tocopherol decreased at calving for cows fed dietary treatments with low or intermediate concentrations of vitamin E, but not for cows fed the high vitamin E treatment. High dietary vitamin E increased concentrations of alpha-tocopherol in blood neutrophils at parturition, but no difference was found for the other two treatments. The percentage of quarters with new infections at calving was not different (32.0%) between cows receiving treatments that contained low and intermediate concentrations of vitamin E but was reduced (11.8%) in cows receiving the high vitamin E treatment. Clinical mastitis affected 25.0, 16.7, and 2.6% of quarters during the first 7 d of lactation for cows receiving the low, intermediate, and high vitamin E treatments, respectively. Cows with plasma concentrations of alpha-tocopherol < 3.0 micrograms/ml at calving were 9.4 times more likely to have clinical mastitis during the first 7 d of lactation than were cows with plasma concentrations of alpha-tocopherol > 3.0 micrograms/ml.

Dietary vitamin E and selenium affect mastitis and milk quality.

Vitamin E and selenium (SE) are essential nutrients that are integral components of the antioxidant defense of tissues and cells. Soils in many of the important dairy regions of the world are Se-deficient, and feedstuffs grown on these soils will not provide adequate dietary Se. Cattle consuming stored forages are likely to be low in vitamin E unless supplemented, and vitamin E deficiencies are frequently observed in peripartum dairy cows. Many new intramammary infections (IMI) occur in the 2 wk before and after calving. Deficiencies of either vitamin E or Se have been associated with increased incidence and severity of IMI, increased clinical mastitis cases, and higher somatic cell counts (SCC) in individual cows and bulk tank milk. Somatic cell counts are a primary indicator of mastitis and milk quality in dairy herds. The polymorphonuclear neutrophil (PMN) is a major defensive mechanism against infection in the bovine mammary gland. A know consequence of vitamin E and Se deficiency is impaired PMN activity and postpartum vitamin E deficiencies are frequently observed in dairy cows. Dietary supplementation of cows with Se and vitamin E results in a more rapid PMN influx into milk following intramammary bacterial challenge and increased intracellular kill of ingested bacteria by PMN. Subcutaneous injections of vitamin E approximately 10 and 5 d before calving successfully elevated PMN alpha-tocopherol concentrations during the periparturient period and negated the suppressed intracellular kill of bacteria by PMN that commonly is observed around calving.

Isotype-specific antibody responses to rotavirus and virus proteins in cows inoculated with subunit vaccines composed of recombinant SA11 rotavirus core-like particles (CLP) or virus-like particles (VLP).

The isotype antibody responses to bovine IND P5, G6 and simian SA11 P2, G3 rotavirus and SA11 rotavirus proteins (VP4, VP6 and VP7) in serum, colostrum and milk were analysed by ELISA in three groups of vaccinated cows and nonvaccinated controls. Pregnant cows were vaccinated intramuscularly and intramammarily with recombinant baculovirus-expressed SA11 rotavirus VLP (triple-layered virus-like particles containing rotavirus VP2, VP4, VP6 and VP7); CLP (double-layered core-like particles containing rotavirus VP2 and VP6); or inactivated SA11 rotavirus, respectively. Rotavirus antigen titers were highest (30-200-fold) in ELISA in the VLP vaccine compared to the inactivated SA11 vaccine. The IgG1, IgG2 and IgM geometric mean antibody titers (GMT) to rotavirus (titers to bovine rotavirus vs SA11 rotavirus did not differ significantly for any isotype or group) and the IgG2 GMT to VP6 in serum at calving in the vaccinated groups were significantly (P < 0.05) higher than in the control group. In colostrum, IgG1 and IgA rotavirus antibody titers were significantly elevated for VLP (IgG1 GMT 832225; IgA GMT 16384), CLP (IgG1 GMT 660561; IgA GMT 10321) and SA11 (IgG1 GMT 131072; IgA GMT 1448) vaccinated cows compared to control cows (IgG1 GMT 11585; IgA GMT 45). The IgG1 and IgA GMT to rotavirus were significantly elevated (6-100-fold) in milk of VLP and CLP vaccinated cows compared to SA11 vaccinated or control cows. The isotype antibody responses to VP6 in serum, colostrum and milk paralleled the responses to rotavirus, but titers were approximately 2-10-fold lower. Only cows vaccinated with VLP had significantly enhanced serum, colostral and milk antibody titers to rotavirus VP4 and VP7. These results demonstrate that rotavirus antibody titers in serum, colostrum and milk are significantly enhanced by use of non-infectious VLP, CLP and inactivated SA11 rotavirus vaccines, but the VLP or CLP vaccines induced the highest antibody responses, corresponding to their higher rotavirus antigen titers measured by ELISA.

alpha-Tocopherol concentrations in milk and plasma during clinical Escherichia coli mastitis.

Eighteen cows were challenged by intramammary infusion with Escherichia coli 727 to determine the effects of acute clinical mastitis on alpha-tocopherol concentrations in plasma and milk. Cows were fed diets supplemented with 1000 IU of vitamin E/d from calving through the experimental period. At challenge, geometric mean DIM was 33 d. Each mammary quarter was diagnosed with an IMI and clinical mastitis at 24 and 48 h after challenge. The alpha-tocopherol concentrations in milk from challenged quarters were approximately 60% greater by 24 and 48 h after challenge than concentrations at prechallenge and 168 h postchallenge. Plasma alpha-tocopherol concentrations did not change after intramammary challenge. The alpha-tocopherol in plasma and milk was correlated at 48 and 168 h postchallenge but not at prechallenge or 24 h postchallenge. Milk alpha-tocopherol and SCC were correlated positively across all sample periods. Milk fat and milk alpha-tocopherol concentrations were correlated at each sample period except 24 h postchallenge. Increases in milk alpha-tocopherol during clinical mastitis were not correlated to milk production, DMI, or BSA concentration in milk. Changes in milk alpha-tocopherol concentration during clinical mastitis were similar to the dynamics of milk SCC.

Factors affecting serum selenium and vitamin E concentrations in dairy cows.

Supplementation of selenium and vitamin E to enhance disease resistance in dairy cattle has become common, particularly to prevent periparturient reproductive disorders and mastitis. To establish reference values for serum vitamin E and selenium concentrations in postparturient dairy cattle and to determine whether serum concentrations of these micronutrients varied with season and stage of lactation, cows from a stratified random sample of 50 herds were studied for 1 year. Blood samples were collected from each of the 50 study herds twice, from the 10 most recently parturient cows or from 10% of the herd, whichever was greatest. Mean concentration of vitamin E and selenium was 2.55 micrograms/ml and 78.12 ng/ml, respectively. Vitamin E concentrations were significantly (P < 0.05) higher during the summer and fall than during the winter and spring. Selenium concentrations were significantly (P < 0.05) lower during the summer and fall than during the winter and spring. Herd, season of blood sample collection, and time since parturition were significant (P < 0.02) in explaining variation in vitamin E and selenium concentrations.

Effect of dietary fat and vitamin E on alpha-tocopherol and beta-carotene in blood of peripartum cows.

Nonlactating cows were fed diets containing 88% grass forage and 12% concentrate (DM basis). Starting 14 d prior to anticipated calving, the concentrate was changed to provide 0 or 200 g of supplemental fat and 0 or 890 IU of supplemental vitamin E daily. Following parturition, cows were fed for 14 d a 50% concentrate diet that provided the same amounts of supplemental fat and vitamin E that were fed during the dry period. Plasma was sampled and analyzed for alpha-tocopherol and beta-carotene. Supplemental dietary fat elevated plasma concentrations of both nutrients during the peripartum period. Supplemental dietary vitamin E elevated concentrations of alpha-tocopherol during that period. Dietary fat also increased concentrations of plasma cholesterol. When alpha-tocopherol and beta-carotene were expressed per unit of plasma cholesterol, fat supplementation did not affect concentrations. Dietary treatments did not influence concentrations of alpha-tocopherol in blood neutrophils and did not affect intracellular kill of bacteria by neutrophils. Essentially no beta-carotene was found in the neutrophils. For colostrum, dietary vitamin E increased concentrations of alpha-tocopherol and decreased concentrations of beta-carotene.

Role of vitamin E and selenium in host defense against mastitis.

Vitamin E and Se are essential nutrients that share common biological activities. Deficiencies in either of these micronutrients have been related in increased incidence and severity of mastitis. A known physiological consequence of alpha-tocopherol or Se deficiency is reduced neutrophil activity. Vitamin E and the Se-containing enzyme, glutathione peroxidase, and antioxidants that protect neutrophils from the destructive action of toxic oxygen molecules necessary for intracellular kill of ingested pathogens. Dietary supplementation of cattle with Se results in a more rapid neutrophil influx into milk following intramammary bacterial challenge and increased intracellular kill of ingested bacteria by neutrophils. Dietary supplementation of early lactation cows with vitamin E results in increased bactericidal activity by bovine blood neutrophils. Recently completed trials have shown that subcutaneous injections of vitamin E approximately 10 and 5 d prior to calving successfully elevated neutrophil alpha-tocopherol concentrations during the periparturient period and negated the suppressed intracellular kill of bacteria by neutrophils that is commonly observed at calving.

Vitamin E as an adjuvant in an Escherichia coli J5 vaccine.

Vitamin E was tested as an adjuvant in an Escherichia coli (O111:B4) J5 vaccine. Twenty cows were assigned to five groups of 4 cows. Cows in four groups were vaccinated with an E. coli J5 bacterin containing 5 ml of 10(9) boiled cells/ml. Vaccinations were at drying off, 30 d after drying off, and within 48 h after calving. Vaccine adjuvants differed among groups. The four treatment adjuvants were 5 ml of Freund's incomplete adjuvant, 5 ml of vitamin E, 2.5 ml of Freund's plus 2.5 ml of vitamin E, and 5 ml of PBS. Cows in the fifth group were unimmunized controls. A front mammary quarter of each cow was challenged by infusion of 10 micrograms of E. coli J5 lipopolysaccharide approximately 4 wk into lactation. Vitamin E alone enhanced serum IgM titers but had no effect on milk IgM or serum and milk IgG titers. The mixture of Freund's plus vitamin E resulted in peak IgG titers in serum and milk comparable with that of Freund's alone. Persistency of IgG titers in cows immunized with the Freund's plus vitamin E mixture was greater than the persistency of titers for cows immunized with the vaccine containing Freund's alone as the adjuvant. The mixture of Freund's plus vitamin E had a synergistic effect in reducing severity of systemic clinical signs.

Effect of supplementing periparturient cows with vitamin E on distribution of alpha-tocopherol in blood.

Holstein cows were fed 0 or 1000 IU/d of supplemental vitamin E during the dry period and injected with placebo or 3000 IU of vitamin E at 10 and 5 d prior to anticipated calving. Blood was collected at various times, starting at 60 d prepartum (dry off) and concluding 30 d postpartum, and separated into plasma, red blood cells, and neutrophils. Oral supplementation increased concentrations of alpha-tocopherol in plasma and red blood cells on 10 d, but not on 5 d prepartum. Injection of vitamin E increased alpha-tocopherol in plasma, red blood cells, and neutrophils at d 5 prior to and on the day of parturition. Concentrations of alpha-tocopherol and cholesterol in plasma were correlated, and both were at their nadir at parturition. Concentrations of alpha-tocopherol in plasma and red blood cells were correlated, but the correlation increased when plasma alpha-tocopherol was expressed per unit of cholesterol. alpha-Tocopherol in plasma and neutrophils had a low correlation. Injection of alpha-tocopherol increased its concentration in plasma, red blood cells, and neutrophils during the periparturient period. Concentration of alpha-tocopherol in plasma of periparturient cows may be limited by its low lipid content, and transport mechanisms for alpha-tocopherol may differ between red blood cells and neutrophils.

Prevention of human rotavirus-induced diarrhea in gnotobiotic piglets using bovine antibody.

The efficacy of passively administered bovine antibody for preventing human rotavirus (HRV)-induced diarrhea was investigated using a gnotobiotic pig model. Cows were immunized with inactivated HRV serotypes 1 (Wa) and 2 (S2) and simian rotavirus serotype 3 (SA11), and immune colostrum and milk were collected. Antibody concentrates derived from these materials were fed to germ-free piglets that were subsequently inoculated with HRV Wa. Both viral shedding and diarrhea were effectively reduced or eliminated in a dose-dependent manner as a result of HRV immune antibody feeding. A quantitative virus-neutralizing (VN) antibody method permitted assessment of the functional antibody dose required to achieve a 50% reduction of disease (PD50). PD50 dose levels of 15.8 and 19.5 x 10(6) VN antibody units were determined for inhibition of diarrhea and viral shedding, respectively. Studies reported here provide new information on the quantitative relationship between protective antibody dose and diarrheal disease response.

Bovine neutrophil responses to parenteral vitamin E.

Twenty-eight Holsteins were tested to determine effects of dietary and parenteral vitamin E supplementation during the dry period on plasma alpha-tocopherol and in vitro neutrophil functions at calving. Cows were assigned to one of four experimental groups receiving either supplemental dietary vitamin E, injections of vitamin E, both dietary and injections of vitamin E, or neither source of supplemental vitamin E during the dry period in a 2 x 2 factorial arrangement. Cows receiving parenteral vitamin E were injected subcutaneously with 3000 IU of vitamin E (dl-alpha-tocopherol) at 10 and 5 d prior to anticipated calving. Cows not receiving parenteral vitamin E were injected with a placebo. Experimental groups receiving dietary vitamin E during the dry period were supplemented with 1040 IU/d compared with none for controls. Cows injected with vitamin E had greater plasma alpha-tocopherol concentration 5 d after the first injection, at calving, and 1 wk after calving than did cows injected with placebo. Plasma alpha-tocopherol concentrations did not differ between dietary vitamin E treatment groups from calving through 4 wk postpartum. No interaction was found between dietary and parenteral supplementation of vitamin E on plasma alpha-tocopherol concentration. Neutrophils from cows injected with vitamin E had greater intracellular kill of bacteria at calving than did neutrophils from placebo-injected cows. Neither phagocytic index nor percentage of neutrophils phagocytizing differed between vitamin E-injected and placebo-injected cows. Dietary vitamin E during the dry period had no effect on neutrophil function at calving. Intracellular kill and plasma alpha-tocopherol were correlated at calving.

Opsonic activity of bovine serum and mammary secretion after Escherichia coli J5 vaccination.

Six pairs of cows were used to determine the effects of immunization with an Escherichia coli (O111:B4) J5 bacterin on in vitro opsonization of a smooth heterologous strain of E. coli. One cow in each pair was either immunized with the vaccine or sham-immunized at drying off, 30 d after drying off, and at calving. Opsonizing bacteria with serum collected from vaccinated cows 21 d after calving resulted in higher mean number of intracellular bacteria per phagocytosing neutrophil than opsonizing bacteria with serum collected from control cows. Phagocytic parameters using serum collected at drying off and calving did not differ between treatment groups. A trend for enhanced opsonic activity of colostrum from vaccinates was noted. Enhanced opsonization by serum from vaccinated cows coincided with higher serum IgM titer to E. coli J5 whole cell antigen compared with controls. Serum IgG titers to E. coli J5 did not differ between groups. Colostrum IgG titers to E. coli J5 were greater at calving in vaccinated than in control cows. Colostrum and milk collected 21 d after calving from vaccinated cows had higher IgM titers to E. coli J5 than did mammary secretions from control cows. Numbers of intracellular bacteria per phagocytizing neutrophil were correlated positively with IgM titers to E. coli J5 in both serum and colostrum.