Rift Valley fever MP-12 vaccine elicits an early protective immune response in mice.

Rift Valley fever virus (RVFV) is an important mosquito-borne pathogen that causes outbreaks of severe disease in people and livestock throughout Africa and the Arabian Peninsula. The development of an effective veterinary and human vaccine to protect against Rift Valley fever (RVF) disease remains a high priority. The live attenuated RVFV MP-12 is a promising vaccine candidate for the prevention of RVF in both human and domestic ruminants. The aim of this study was to determine the onset of protective immunity elicted in mice by a single dose of this vaccine. Groups of CD-1 mice were vaccinated intraperitoneally with RVFV MP-12 vaccine and challenged on days 2, 5, 6 and 7 post-vaccination (PV) with a lethal dose of virulent RVFV. The mice were observed once daily for terminal morbidity and blood samples were obtained from the retro-orbital sinus complex on days 23 and 28 PV of surviving mice to determine RVFV neutralizing antibody titers. In one test, 2 of 3 mice challenged on day 2 PV survived and all 3 mice challenged at days 5 and 7 PV also survived. A second test of 10 mice per group was performed, and half (5) of those challenged at day 2 PV survived while all (10) survived challenge at day 4 and 6 PV. All surviving animals develop antibody that ranged from 1:80 to 1:1,280 PV. In a separate experiment, RVFV MP-12 vaccinated CD-1 mice, but not challenged developed a low viremia for the first 3 days PV and neutralzing antibody was detected on days 5 through day 28 PV. These findings demonstrated that the RVFV MP-12 vaccine elicited a rapid protective immune response in mice as early as 2 days PV, thus further supporting the effectiveness of this vaccine candidate for preventing RVF among humans and domestic ruminants.

Glucose and acetate metabolism in bovine intramuscular and subcutaneous adipose tissues from steers infused with glucose, propionate, or acetate.

We hypothesized that abomasal infusion of glucose would promote de novo fatty acid biosynthesis from glucose in vitro in bovine intramuscular (i.m.) and subcutaneous (s.c.) adipose tissues to a greater extent than ruminal infusion of acetate, propionate, or glucose. Angus crossbred steers (n = 24), 22 mo of age, were fitted with ruminal cannulas, and steers were adapted to another corn/sorghum finishing diet over a 2-wk period while recovering from the placement of the cannulas. After the adaptation period, the steers were fed the second finishing diet at 130% of their voluntary intake and were infused with isocaloric amounts (3.76 Mcal/d) of glucose, propionate, or acetate for 35 d. Glucose was infused either into the rumen or into the abomasum, whereas propionate and acetate were infused into the rumen. Acetate infusion decreased DM and DE intakes (P < 0.05). The 5th to 8th longissimus muscle section was removed immediately and transported to the laboratory within 10 min post-exsanguination in 38 °C, oxygenated Krebs Henseleit buffer containing 5 mM glucose and 5 mM acetate. Intramuscular and s.c. adipose tissues were dissected from the muscle and incubated in vitro in 5 mM glucose plus 5 mM acetate (containing [U-14C]glucose or [1-14C]acetate). Lipid content was lower (P = 0.04) in i.m. adipose tissue of the acetate-infused steers than in the other treatment groups, and i.m. adipocytes from acetate-infused steers were smaller (P = 0.01) than those from propionate-infused steers. The rate of incorporation of acetate into glyceride-fatty acids (GFA) in i.m. and s.c. adipose tissues was greater (P < 0.03) in steers receiving ruminal or abomasal infusions of glucose than in adipose tissues from steers infused with acetate. The greatest rates of GFA synthesis were observed in s.c. adipose tissue from steers infused ruminally with propionate or abomasally infused with glucose (P < 0.001). In i.m. and s.c. adipose tissues, the proportion of acetyl units from acetate incorporated into GFA was greater in steers receiving glucose infusion in the rumen or abomasum than in steers receiving acetate or propionate infusion (P < 0.05). Contrary to our hypothesis, abomasal glucose infusion did not promote greater fatty acid biosynthesis from glucose in i.m. adipose tissue than ruminal glucose infusion. However, glucose infusion caused the greatest production of acetyl units from acetate in i.m. and s.c. adipose tissues.

The influence of taste in willingness-to-pay valuations of sirloin steaks from postextraction algal residue-fed cattle.

Consumer preferences and willingness-to-pay (WTP) for beef sirloin steaks with differing production, physical, and credence attributes related to the use of postextraction algal residue (PEAR), a novel feed ingredient, were estimated. Ninety-six consumers participated in a sensory tasting panel before completing a choice set survey; 127 consumers completed only the choice set survey without sampling products. Steaks from grain- and PEAR-fed steers had similar Warner-Bratzler shear force (WBSF) scores (1.89 kg and 2.01 kg, respectively; = 0.77) and had lower WBSF scores than steaks from grass-fed steers (3.37 kg; < 0.05). Eicosapentaenoic acid (20:5) was not different among steaks from grain- and PEAR-fed steers ( = 0.39) but was greater compared with steaks from grass-fed cattle ( ≤ 0.03). Panelists in the sensory portion of the study evaluated beef samples for like/dislike of overall sample, overall flavor, beefy flavor, and juiciness. Panelist rating of overall like, overall flavor like, and beefy flavor like were not different between the PEAR- and grain-fed treatments ( > 0.26). Panelists rated the juiciness like/dislike of steaks from PEAR-fed cattle the highest ( < 0.01) among the 3 samples. Sensory tasting of the products was observed to alter the preferences of consumers. Consumers who completed only the survey negatively perceived beef from PEAR-fed cattle compared with beef from grain-fed cattle, with a WTP discount of -US$1.17/kg. However, with sensory tasting, the WTP for beef from PEAR-fed cattle was not discounted relative to beef from grain-fed cattle ( = 0.21). The nontasting consumers had much higher stated WTP values for credence attributes. Factors that influence the eating experience (tenderness and quality grade) dominated as the most important attributes on WTP among the tasting group. The use of no hormones and no antibiotics in production had a premium of $2.34/kg among the nontasting group, but with tasting, the premium was $1.19/kg. If PEAR-fed beef came to market, there would be no need to differentiate it from grain-fed beef unless retailers wanted to market it as a differentiated product. If it were marketed as a differentiated product, retailers would need to hold promotional tastings to change consumer's preconceived notions about the product.

Pathogenicity and neurovirulence of a mutagen-attenuated Rift Valley fever vaccine in rhesus monkeys.

Rhesus macaques, intravenously inoculated with virulent Rift Valley fever virus, develop viremia and biochemical evidence of liver damage and serve as a model for human disease. Some of these monkeys suffer more serious disease with hemorrhagic phenomena and approximately 20% die with frank hemorrhage. Presently, the only Rift Valley fever vaccine approved for use in humans is a formalin-killed product that requires annual booster vaccinations. Efforts to produce an improved vaccine to replace the present vaccine have led to a mutagen-attenuated strain of Rift Valley fever virus that was found to be markedly attenuated for rhesus macaques and showed promise as a vaccine candidate for human use. Neurovirulence testing in rhesus monkeys showed that, while the vaccine was not completely innocuous, residual lesions were no more severe than the currently used 17D yellow fever vaccine.

Quantum mechanical quantitative structure activity relationships to avoid mutagenicity in dental monomers.

The objective of this study was to identify through quantum mechanical quantitative structure activity relationships (Q-QSARs) chemical structures in dental monomers that influence their mutagenicity. AMPAC, a semiempirical computer program that provides quantum mechanical information for chemical structures, was applied to three series of reference chemicals: a set of methacrylates, a set of aromatic and a set of aliphatic epoxy compounds. QSAR models were developed using this chemical information together with mutagenicity data (Salmonella TA 100, Ames Test). CODESSA, a QSAR program that calculates quantum chemical descriptors from information generated by AMPAC and statistically matches these descriptors with observed biological properties was used. QSARs were developed which had r2 values exceeding 0.90 for each study series. These QSARs were used to accurately predict the mutagenicity of BISGMA. a monomer commonly used in dentistry, and two epoxy monomers with developing use in dentistry, GY-281 and UVR-6105. The Q-QSAR quantum mechanical descriptors correctly predicted the level of mutagenicity for all three compounds. The descriptors in the correlation equation pointed to components of structure that may contribute to mutagenesis. The QSARs also provided 'dose windows' for testing mutagenicity, circumventing the need for extensive dose exploration in the laboratory. The Q-QSAR method promises an approach for biomaterials scientists to predict and avoid mutagenicity from the chemicals used in new biomaterial designs.

COOH-terminal truncated alpha(1S) subunits conduct current better than full-length dihydropyridine receptors.

Skeletal muscle dihydropyridine (DHP) receptors function both as voltage-activated Ca(2+) channels and as voltage sensors for coupling membrane depolarization to release of Ca(2+) from the sarcoplasmic reticulum. In skeletal muscle, the principal or alpha(1S) subunit occurs in full-length ( approximately 10% of total) and post-transcriptionally truncated ( approximately 90%) forms, which has raised the possibility that the two functional roles are subserved by DHP receptors comprised of different sized alpha(1S) subunits. We tested the functional properties of each form by injecting oocytes with cRNAs coding for full-length (alpha(1S)) or truncated (alpha(1SDeltaC)) alpha subunits. Both translation products were expressed in the membrane, as evidenced by increases in the gating charge (Q(max) 80-150 pC). Thus, oocytes provide a robust expression system for the study of gating charge movement in alpha(1S), unencumbered by contributions from other voltage-gated channels or the complexities of the transverse tubules. As in recordings from skeletal muscle, for heterologously expressed channels the peak inward Ba(2+) currents were small relative to Q(max). The truncated alpha(1SDeltaC) protein, however, supported much larger ionic currents than the full-length product. These data raise the possibility that DHP receptors containing the more abundant, truncated form of the alpha(1S) subunit conduct the majority of the L-type Ca(2+) current in skeletal muscle. Our data also suggest that the carboxyl terminus of the alpha(1S) subunit modulates the coupling between charge movement and channel opening.

Effects of mutations causing hypokalaemic periodic paralysis on the skeletal muscle L-type Ca2+ channel expressed in Xenopus laevis oocytes.

1. A truncated form of the rabbit alpha1S Ca2+ channel subunit (alpha1SDeltaC) was expressed with the beta1b, alpha2delta and gamma auxiliary subunits in Xenopus laevis oocytes. After 5-7 days, skeletal muscle L-type currents were measured (469 +/- 48 nA in 10 mM Ba2+). All three of the auxiliary subunits were necessary to record significant L-type current. A rapidly inactivating, dihydropyridine-insensitive endogenous Ba2+ current was observed in oocytes expressing the auxiliary subunits without an exogenous alpha subunit. Expression of full-length alpha1S gave 10-fold smaller currents than the truncated form. 2. Three missense mutations causing hypokalaemic periodic paralysis (R528H in domain II S4 of the alpha1S subunit; R1239H and R1239G in domain IV S4) were introduced into alpha1SDeltaC and expressed in oocytes. L-type current was separated from the endogenous current by nimodipine subtraction. All three of the mutations reduced L-type current amplitude ( approximately 40 % for R528H, approximately 60-70 % for R1239H and R1239G). 3. The disease mutations altered the activation properties of L-type current. R528H shifted the G(V) curve approximately 5 mV to the left and modestly reduced the voltage dependence of the activation time constant, tauact. R1239H and R1239G shifted the G(V) curve approximately 5-10 mV to the right and dramatically slowed tauact at depolarized test potentials. 4. The voltage dependence of steady-state inactivation was not significantly altered by any of the disease mutations. 5. Wild-type and mutant L-type currents were also measured in the presence of (-)-Bay K8644, which boosted the amplitude approximately 5- to 7-fold. The effects of the mutations on the position of the G(V) curve and the voltage dependence of tauact were essentially the same as in the absence of agonist. Bay K-enhanced tail currents were slowed by R528H and accelerated by R1239H and R1239G. 6. We conclude that the domain IV mutations R1239H and R1239G have similar effects on the gating properties of the skeletal muscle L-type Ca2+ channel expressed in Xenopus oocytes, while the domain II mutation R528H has distinct effects. This result implies that the location of the substitutions is more important than their degree of conservation in determining their biophysical consequences.

Isolation of a single carboxyl-carboxylate proton binding site in the pore of a cyclic nucleotide-gated channel.

The pore of the catfish olfactory cyclic nucleotide-gated (CNG) channel contains four conserved glutamate residues, one from each subunit, that form a high-affinity binding site for extracellular divalent cations. Previous work showed that these residues form two independent and equivalent high-pKa (approximately 7.6) proton binding sites, giving rise to three pH-dependent conductance states, and it was suggested that the sites were formed by pairing of the glutamates into two independent carboxyl-carboxylates. To test further this physical picture, wild-type CNG subunits were coexpressed in Xenopus oocytes with subunits lacking the critical glutamate residue, and single channel currents through hybrid CNG channels containing one to three wild-type (WT) subunits were recorded. One of these hybrid channels had two pH-dependent conductance states whose occupancy was controlled by a single high-pKa protonation site. Expression of dimers of concatenated CNG channel subunits confirmed that this hybrid contained two WT and two mutant subunits, supporting the idea that a single protonation site is made from two glutamates (dimer expression also implied the subunit makeup of the other hybrid channels). Thus, the proton binding sites in the WT channel occur as a result of the pairing of two glutamate residues. This conclusion places these residues in close proximity to one another in the pore and implies that at any instant in time detailed fourfold symmetry is disrupted.

Utilization of bioelectrical impedance to predict carcass composition of Holstein steers at 3, 6, 9, and 12 months of age.

The objective of this experiment was to study the usefulness of bioelectrical impedance analysis (BIA) in determining soft tissue composition (STC) and carcass fat-free mass (CFFM) of Holstein steers at different ages. Growth data and prediction of STC and CFFM were determined for four groups of Holstein steers: 12 of 3 mo, 12 of 6 mo, 15 of 9 mo, and 16 of 12 mo of age. Average weight for animals at 3, 6, 9, and 12 mo were 96.6, 204.7, 354.1, and 465.9 kg, respectively. Average fat content of carcass soft tissue at 3, 6, 9, and 12 mo were 2.6, 9.8, 18.2, and 24.6%, respectively. Average protein content of the carcass soft tissue was 20.7% at 3 mo, 20% at 6 mo, 18.30% at 9 mo, and 16.9% at 12 mo of age. Feed and water were withheld for 20 h before the BIA was applied. Steers were sedated and forced to recumbency in a lateral position on their right sides over a nonconductive surface. Two electrodes were placed on each limb of the right side (metatarsal and metacarpal regions on back and front foot, respectively). Resistance (Rs) and reactance (Xc) were obtained by attaching four terminals to the electrodes. Impedance and other predictors such as Vol1 (L/Rs), Vol2 (L2/(RS2+Xc2).5, Vol3 (geometrical animal volume), L (2 x height + body length), and L2 were calculated from Rs and Xc, and body measurements and were used to generate prediction equations for CFFM and carcass soft tissue composition. Carcass fat-free mass was predicted accurately for all age groups and the pooled data (r2 = .99 at 3 mo, .99 at 6 mo, .97 at 9 mo, .77 at 12 mo, and .98 for the pooled data). Correlation coefficients between impedance readings and CFFM and carcass composition were calculated. Carcass CFFM and kilograms of H2O for the pooled data (across age groups) were both correlated highly to Vol1 (.97), Vol2 (.95), L (.97), and L2 (.97).

Gating of the L-type Ca channel in human skeletal myotubes: an activation defect caused by the hypokalemic periodic paralysis mutation R528H.

The skeletal muscle L-type Ca channel serves a dual role as a calcium-conducting pore and as the voltage sensor coupling t-tubule depolarization to calcium release from the sarcoplasmic reticulum. Mutations in this channel cause hypokalemic periodic paralysis (HypoPP), a human autosomal dominant disorder characterized by episodic failure of muscle excitability that occurs in association with a decrease in serum potassium. The voltage-dependent gating of L-type Ca channels was characterized by recording whole-cell Ca currents in myotubes cultured from three normal individuals and from a patient carrying the HypoPP mutation R528H. We found two effects of the R528H mutation on the L-type Ca current in HypoPP myotubes: (1) a mild reduction in current density and (2) a significant slowing of the rate of activation. We also measured the voltage dependence of steady-state L-type Ca current inactivation and characterized, for the first time in a mammalian preparation, the kinetics of both entry into and recovery from inactivation over a wide range of voltages. The R528H mutation had no effect on the kinetics or voltage dependence of inactivation.

Effects of form of the diet on anatomical, microbial, and fermentative development of the rumen of neonatal calves.

Eight neonatal, Holstein bull calves were paired by birth date and birth weight and randomly assigned to either a finely ground or unground control diet (chopped hay and rolled grain) to study the effects of the physical form of the diet on anatomical, microbial, and fermentative development of the rumen. The diets varied in particle size but were identical in composition (25% alfalfa hay and 75% grain mix). Calves were fed milk at 8% of birth weight daily until weaning. Feed intake was equalized for each pair of calves. Ruminal fluid samples were collected from ruminal cannulas to determine pH, fermentation products, and buffering capacity and to enumerate bacteria. Calves were slaughtered at 10 wk of age, and weights of the full and empty reticulorumen, abomasum, and omasum were recorded. Ruminal tissue samples were taken to assess papillary development by morphometric measurements. Calves had similar body weights at wk 10. Ruminal pH was affected by age and was lower for calves fed the ground diet. Total anaerobic bacterial counts were not affected by the physical form of the diet; however, calves fed the ground diet had lower numbers of cellulolytic bacteria and higher numbers of amylolytic bacteria than did calves fed the unground diet. Physical form of the diet did not affect the weights of the reticulorumen whether full or empty. However, calves fed the ground diet had heavier omasum weights, both full and empty. Physical form of the diet affected papillary size and shape but did not influence the muscle thickness of rumen. Results indicated that the physical form of the diet had a significant influence on the anatomical and microbial development of the forestomac and, therefore, might influence future performance.

Cells are added to the archenteron during and following secondary invagination in the sea urchin Lytechinus variegatus.

In the present investigation, nuclei of endodermal cells, primary and secondary mesenchyme cells (PMCs and SMCs), and small micromere descendants (SMDs) of the sea urchin Lytechinus variegatus were counted and mapped at five developmental stages, ranging from primary invagination to pluteus larva. The archenteron and its derivatives were measured three dimensionally with STERECON analytical software. For the first time SMC production is included in the kinetic analysis of archenteron formation. While the archenteron lumen doubled in length during secondary invagination, the number of archenteron cells increased by at least 38% (over 50% when SMCs that emigrated from the tip of the archenteron were included). The volume of the archenteron epithelial wall plus the volume of 17 new SMCs increased by 40% over the equivalent volumes at the end of primary invagination. Because secondary invagination involves the addition of archenteron cells and an increase in volume of the archenteron epithelium, we conclude that secondary invagination is not accomplished simply by the rearrangement and reshaping of the primary archenteron cells. Both archenteron cell number and wall volume continued to increase at the same rates from the end of secondary invagination until the 27-h prism stage, although the lumen lengthened more slowly. SMCs were also produced at a constant rate from primary invagination until the prism stage. Because the production of both endodermal and mesodermal cells continues until the late prism stage, we conclude that gastrulation (defined as the establishment of the germ layers) also extends into the late prism stage.

A new method of measuring diet abrasion and its effect on the development of the forestomach.

Twelve newborn Holstein bull calves were used to evaluate the effects of dietary abrasiveness, determined by a new method, on ruminal development. Calves were blocked by age and body weight and were assigned to one of three different diets. Each diet had the same ingredients but different particle sizes, which resulted in different abrasive values. No differences were detected in molar percentages of volatile fatty acids in ruminal fluid or in plasma concentrations of urea, glucose, or beta-hydroxybutyrate. The pH of ruminal fluid was lower for calves fed the fine and intermediate diets than for those fed the coarse diet. Digesta-free weights of the stomach and stomach compartments were similar among calves fed the three diets, except that omasum weights were heavier for calves fed the fine diet. Length of the ruminal papillae increased as the abrasive value of the diet decreased. Measurements of ruminal tissue layers from the ventral floor of the cranial sac were not different among diets, but the keratin portion represented more of the epithelial layer for calves fed the diet with the lowest abrasive value, thus decreasing the percentage of metabolically active tissue for those calves.

Using dry feed intake as a percentage of initial body weight as a weaning criterion.

Newborn Holstein heifers (n = 32) and bulls (n = 12) were used to investigate the use of dry feed intake as a percentage of birth weight as a weaning criterion. Three different percentages (1, 1.5, and 2%) were used. Calves in the 1% treatment group met the weaning criterion earlier than did those in the 1.5 and 2% treatment groups; no difference was detected between the latter two groups. Total dry feed intake at 8 wk was higher for calves in the 1% treatment group than for calves in the other treatment groups; no difference was detected between the 1.5 and 2% treatment groups. Weights for all calves at 8 wk and weights of heifer calves at 12, 16, and 20 wk were not different among groups. Using dry feed intake at 1% of birth weight as a weaning criterion reduced days to weaning, increased dry feed intake from birth to 8 wk, decreased variation in weaning age, and had no apparent negative effect on growth at 20 wk of age. Using dry feed intake as a percentage of birth weight appears to be a suitable criterion to determine when to wean dairy calves.

Safety and efficacy of a mutagen-attenuated Rift Valley fever virus vaccine in cattle.

OBJECTIVE: To examine safety and efficacy of a mutagen attenuated Rift Valley fever virus (RVFV) vaccine (RVF MP-12) in cattle. ANIMALS: 38 pregnant cows, 14 steers, and 10 lactating dairy cows. PROCEDURE: Pregnant cows in their third, fifth, or eighth month of gestation were vaccinated (1 ml of RVF MP-12 containing 5 log10 plaque-forming units [PFU] of virus) and were monitored daily through parturition for signs of disease, viremia, and immunologic response. Additionally, 10 vaccinated pregnant cows were challenge inoculated with virulent RVFV at post-vaccination day (PVD) 30 and were monitored daily for untoward effects. Ten unvaccinated pregnant cows also were challenge inoculated with virulent RVFV and served as challenge controls. Vaccinated lactating dairy cows were monitored for viremia and virus shedding in the milk through PVD 14. Yearling steers were vaccinated to assess their immunologic response to various doses of vaccine and were challenge inoculated with virulent RVFV at PVD 28 to assess protection. RESULTS: 10 of 38 (26.3%) cows vaccinated during pregnancy developed transient postvaccination viremia titer > or = 2.5 log10 PFU/ml of serum. All vaccinated cows delivered live, healthy calves that were RVFV seronegative at birth, but which quickly acquired colostral antibodies. Vaccinated cows and their fetuses were protected when challenge exposed with virulent RVFV at PVD 30, whereas unvaccinated pregnant cows inoculated with RVFV became febrile and viremic, and aborted. Vaccine virus was unsuccessfully sought from milk of lactating dairy cows after vaccination, suggesting that shedding of vaccine virus through milk should not be a concern. Steers, inoculated with tenfold escalating vaccine doses, beginning with 1.0 log10 PFU, were protected against virulent RVFV challenge exposure. CONCLUSIONS: RVF MP-12 may be safe and efficacious for use in pregnant or lactating bovids, and a minimal dose of vaccine may provide suitable protection against viremia.

Safety of a mutagen-attenuated Rift Valley fever virus vaccine in fetal and neonatal bovids.

OBJECTIVE: To examine effects of in utero inoculation with a mutagen-attenuated Rift Valley fever virus (RVFV) vaccine (RVF MP-12) on fetal bovids and to assess the safety and efficacy of calfhood vaccination with RVF MP-12. ANIMALS: 18 pregnant Hereford and Hereford-type cows in the third or fifth month of gestation, their progeny, and 25 calves from cows immunized with RVF MP-12 during pregnancy. PROCEDURE: Bovine fetuses were inoculated, via laparotomy, with 1 ml of RVF MP-12 containing 5 log10 plaque-forming units (PFU) of virus. Blood was obtained from newborn calves prior to their ingestion of colostrum. Immune-naive calves and calves born to RVF MP-12-vaccinated dams, ranging in age from 2 to 45 days, were vaccinated with RVF MP-12, and some were later challenge exposed with 1 ml of 5.7 log10 PFU of virulent RVFV strain ZH-501. Cows were monitored for viremia and antibody responses and for hematologic and serum biochemical alterations through parturition or abortion. RESULTS: Surviving in utero-vaccinated calves were healthy, with no noticeable defects. Except for 1 vaccine-inoculated fetus that died on postinoculation day 21, all in utero-vaccinated fetuses had serum neutralizing antibody titer > or = 1:20 at the time of delivery. All dams of in utero-vaccinated fetuses also developed neutralizing antibody titer. Calves born to cows vaccinated during gestation did not have antibody at birth, and all but 1 quickly acquired colostral antibody. Postparturient inoculation of immune-naive calves and calves with colostral antibodies resulted in no untoward effects, and all calves with detectable neutralizing antibodies were protected against virulent virus challenge exposure. CONCLUSIONS: Fetal death and abortion would be rare even if fetuses were exposed to RVF MP-12. The trauma and complications associated with in utero inoculation do not make this a practical method of immunization. RVF MP-12 was safe, immunogenic, and protective in calves as young as 2 days of age.

Bovine IgG2a antibodies to Haemophilus somnus and allotype expression.

Bovine IgG2a has been implicated in protection against pyogenic infections, including those caused by Haemophilus somnus. To further investigate the role of IgG2a in defense against H. somnus, IgG1 and IgG2a antibodies were purified from antiserum against an immunodominant 40 kDa outer membrane protein (p40) of H. somnus, which was previously shown to passively protect calves against H. somnus pneumonia. The passive protective capacity of anti-p40 IgG1 or IgG2a was evaluated in vivo in calves. Purified anti-p40 IgG1 or IgG2a was incubated with H. somnus for 15 min before intrabronchial inoculation of calves. Bacteria incubated with anti-p40 IgG1 or IgG2a were inoculated into one caudal lung lobe and bacteria incubated with IgG1 or IgG2a from the respective preimmunization serum were inoculated into the contralateral lobe. The volumes of pneumonia in the right and left lungs were determined 24 h later. The difference in volume of pneumonia with H. somnus preincubated in IgG1 pre- and postimmunization anti p40 was less (16 cm3, P = 0.298) than the difference in volume of pneumonia with H. somnus preincubated in IgG2a pre- and postimmunization anti p40 (30 cm3, P = 0.146). Although the differences in lesion size between pre- and postimmunization serum were not statistically significant, the trend suggests IgG2a may be more protective than IgG1. To examine this further, the peptide specificity of these IgG1 and IgG2a antibodies to p40 was examined. After limited proteolysis of p40, IgG2a antibodies reacted with 2 peptides not recognized by IgG1 antibodies. Other peptides were recognized by both isotypes. Since these studies suggested that IgG2a may be important in protection against infection, we then investigated some aspects of the role of the 2 IgG2a allotypes, A1 and A2. In retrospective studies of age differences in expression of IgG2a allotypes, no heterozygotes were detected in calves of 60 d old or less, and fewer heterozygotes were detected in calves 61-120 d old than in cattle older than 270 d (P < 0.01). In a subsequent prospective study of the time course of allotype expression, Holstein calves shown to be heterozygotes expressed the IgG2aA1 allotype early but the IgG2aA2 allotype was not usually detected until 3 to 4 mo of age. Thus, both the retrospective and the prospective studies showed age related differences in expression of the IgG2aA1 and A2 allotypes. This could have implication in protection.

Intake, digestion, and nitrogen utilization by sheep fed tropical legumes with contrasting tannin concentration and astringency.

We conducted an experiment to determine the effects of concentration and astringency of extractable and bound condensed tannins (CT) in tropical legumes on intake, digestibility, and nitrogen (N) utilization by sheep. The test legumes (Desmodium ovalifolium and Flemingia macrophylla) had similar concentrations of extractable CT (90 g/kg DM) but different concentrations of bound CT and astringency of tannins. Chopped, sun-dried forage of each legume was sprayed with either water (control) or polyethylene glycol (PEG, 35 g/kg of DM) to bind extractable CT and fed daily (26 g/kg BW) to eight sheep with ruminal and duodenal cannulas. The sheep also received starch-extracted cassava meal intraruminally (4 g/kg BW) as a constant source of readily fermentable carbohydrates. Intake of the two legumes was not different (P > .05), but it increased an average of 10% (P < .01) when extractable CT were reduced from 90 to 50 g/kg of DM with PEG. Ruminal and total tract digestibilities of OM, NDF, and ADF were greater (P < .01) with D. ovalifolium than with F. macrophylla and increased for both legumes with the addition of PEG. Greater (P < .01) N flow to the duodenum, N absorbed from the intestine, and fecal N were observed with F. macrophylla than with D. ovalifolium. Extraction of CT with PEG resulted in less (P < .05) ruminal escape protein and less (P < .01) fecal N with both legumes, but apparent postruminal N digestion was not affected. Changes in the concentration of extractable CT in tropical legumes can significantly affect forage intake, digestion, and N utilization by sheep.

Bioavailability of alpha-tocopherol fed with retinol and relative bioavailability of D-alpha-tocopherol or DL-alpha-tocopherol acetate.

Two experiments were conducted to examine the effects of the form of alpha-tocopherol or interactions of alpha-tocopherol with vitamin A on its bioavailability. In Experiment 1, Holstein steers were fed a diet that was low in vitamins A and E for 1 mo; then, steers were blocked by body weight (X = 97.5 kg) and assigned randomly to one of four oral treatments: 1) no added vitamins, 2) 442 mg of retinyl acetate, 3) 1342 mg of D-alpha-tocopherol, or 4) 442 mg of retinyl acetate and 1342 mg of D-alpha-tocopherol. Each treatment was given as a pulse dose. Blood was sampled over a 36-h period. Concentrations of plasma retinyl palmitate peaked at 2 to 6 h postsupplementation for all calves and then peaked again at 22 to 28 h for calves receiving vitamin supplements. Concentrations of plasma alpha-tocopherol peaked earliest with D-alpha-tocopherol supplementation alone at 12 to 20 h after supplementation, but simultaneous supplementation with retinyl acetate resulted in lower plasma alpha-tocopherol concentrations. Plasma retinyl palmitate decreased during peak alpha-tocopherol concentrations. In Experiment 2, blood and tissue were analyzed after a single gastric tube administration of a powder (DL-alpha-tocopheryl acetate) or a liquid (D-alpha-tocopherol) form of vitamin E to Holstein calves. Plasma and kidney concentrations of alpha-tocopherol were higher when calves were fed D-alpha-tocopherol than when calves were fed the DL-alpha-tocopherol acetate form. Concentrations in the liver, spleen, adipose tissue, heart, muscle, cellular blood fraction, and gut did not differ between the two forms.

The use of urea dilution for estimation of carcass composition of Holstein steers at 3, 6, 9, and 12 months of age.

Every 3 mo for a 2-yr period, two weaned Holstein steer calves (94.5 kg) were randomly assigned to each of four slaughter age groups (3, 6, 9, and 12 mo). Urea dilution was performed before slaughter, and urea space (US) was calculated as total volume and as a percentage of body weight (BW) and empty body weight (EBW). The relationships between US (kg, % EBW and % BW), BW, and EBW and carcass soft tissue composition (protein, fat, moisture, and ash) were studied. One- and two-pool models were fitted using the urea dilution data and the coefficients of those equations (zero time, A + B), and the intercepts of compartments A and B were used to estimate body volume. Body weight and EBW effectively predicted the amount of water, fat, and protein in the carcass soft tissue. Equations expressed in kilograms were more accurate than those expressed as percentages. Urea space overestimated body water, probably because of the fast rate of urea disappearance in plasma. Correlation coefficients between US and carcass soft tissue water (kg) based on the pooled data ranged from .74 at 6 min to .48 at 42 min after infusion. The biexponential models coefficients explained more of the variation of carcass soft tissue composition than US; correlation coefficients using volume B and the soft tissue composition (in kg) with pooled data were .78 (water), .68 (fat), .69 (ash), and .76 (protein). The relationships between A and soft tissue composition were weaker (water .59, fat .51, ash .58 and protein .59). The highest correlation coefficients were obtained when A + B was used for water, fat, ash, and protein (.83, .70, .74 and .81, respectively). Equations combining BW, EBW, and two-model coefficients (A, B, A + B) explained much of the variation of soft tissue composition. No significant benefit was found in using the urea space at various times after infusion over BW or EBW alone to estimate carcass soft tissue composition in Holstein steers.