Stage of lactation and corresponding diets affect in situ protein degradation by dairy cows.

The influence of stage of lactation and corresponding diets on rates of protein degradation (kd) is largely unstudied. Study objectives were to measure and compare in situ ruminal kd of crude protein (CP) and estimate rumen CP escape (rumen-undegradable protein; RUP) of selected feeds by cows at 3 stages of lactation fed corresponding diets, and to determine the incubation times needed in an enzymatic in vitro procedure, using 0.2 units of Streptomyces griseus protease per percent of true CP, that predicted in situ RUP. Residue CP was measured after in situ fermentation for 4, 8, 12, 24, 36, 48, and 72 h of 5 protein sources and 3 total mixed rations, which were fed to the in situ cows. Two nonlactating (dry) cows and 2 cows each at 190 (mid) and 90 (peak) days of lactation were used. Each pair of cows was offered free-choice diets that differed in composition to meet their corresponding nutrient requirements. Diets had decreasing proportions of forages and contained (dry matter basis) 11.9, 15.1 and 16.4% CP and 54.3, 40.3 and 35.3% neutral detergent fiber, for dry, mid, and peak TMR (TMR1, TMR2, and TMR3), respectively. Intakes were 10.3, 21.4, and 23.8kg of dry matter/d, respectively. Kinetic CP fractions (extractable, potentially degradable, undegradable, or slowly degradable) were unaffected by treatment. Lag time and kd varied among feeds. The kd was faster for all feeds (0.136/h) when incubated in dry-TMR1 cows compared with mid-TMR2 (0.097/h) or peak-TMR3 (0.098/h) cows, and no differences in lag time were detected. Calculated RUP, using estimated passage rates for each cow based on intake, differed between dry-TMR1 (0.382) and mid-TMR2 (0.559) or peak-TMR3 (0.626) cows, with a tendency for mid-TMR2 to be different from peak-TMR3. Using the average kd and lag time obtained from dry-TMR1 to calculate RUP for mid-TMR2 and peak-TMR3 cows using their passage rates reduced RUP values by 6.3 and 9.5 percentage units, respectively. Except for that of herring meal, in vitro residue CP at 6, 12, and 48h of enzymatic hydrolysis was correlated (r=0.90) with in situ RUP of peak-TMR3, mid-TMR2, and dry-TMR1, respectively. Although confounded within treatments, stage of lactation, diet, and intake appeared to affect CP degradation parameters and RUP. Using kd from nonlactating cows, or the RUP calculated from them, may bias diet evaluation or ration formulation for lactating cows. In addition, enzymatic in vitro predictions of RUP should be measured using incubation times that are appropriate for lactating cows.

How do dairy cows chew?--particle size analysis of selected feeds with different particle length distributions and of respective ingested bolus particles.

Not only feed but also respective bolus particle size could alter diet efficiency and cow performance. The objective of this project was to characterize particle size of selected feeds and respective swallowed boli. Feed samples included 6 different particle length rye grass hay samples, 1 grass silage, 1 corn silage, and 1 total mixed ration (TMR). Rye grass hay samples consisted of long hay and chopped hay particles retained on the 19- (19_PSPS hay), 8- (8_PSPS hay), and 1.18-mm (1.18_PSPS hay) Penn State Particle Separator (PSPS) screens and those collected on the pan (PSPS_pan hay). A sixth hay treatment was rye grass forage cut at 50-mm lengths and dried to hay (50-mm hay). Treatments were offered to 4 nonlactating and 4 lactating cows following rumen evacuation. Swallowed boli were collected and the number of chews per gram of ingested feed dry matter was determined. Feed and bolus particles of lengths ≥5mm were collected on a 1.6-mm screen using a horizontal wet sieving technique. This cut point was chosen, as the literature suggests that most fecal particles are shorter than 5mm. Dry matter proportions on this screen (PROP_1.6) were determined and particle lengths of retained particles were measured by image analysis. Mean particle lengths (ML) were calculated considering particles ≥5mm in length. Boli of long hay, of 19_PSPS hay, of 8_PSPS hay, and of 50-mm hay had similar ML of 10 to 11mm. Bolus PROP_1.6 were also similar between these treatments, ranging from 0.54 to 0.69. Bolus particle lengths and distributions of these treatments were not related to respective hay particles. Bolus of 1.18_PSPS hay had PROP_1.6 of 0.51 and a smaller ML of 8mm. The PSPS_pan hay had PROP_1.6 of only 0.33, but was still chewed intensely. Apparently, little particle size reduction occurred when cows ate the TMR or the silages. Feed and respective bolus PROP_1.6 were as follows: 0.66 and 0.59 in grass silage, 0.52 and 0.55 in corn silage, and 0.44 and 0.38 in the TMR. Feed and respective bolus ML were as follows: 13.8 and 11.6mm in grass silage, 12.0 and 11.2mm in corn silage, and 13.1 and 12.5mm in the TMR. Rye grass hay particles retained on PSPS screens ≥8mm, with ML of at least 25mm were longer compared with TMR particles, but respective bolus particles were shorter. Bolus particle size is not associated with the size of large feed particles chewed to a constant size that is appropriate for deglutition. This size may be related to feed chemical composition.

Composition and aroma compounds of Ragusano cheese: native pasture and total mixed rations.

Raw milk from 13 cows fed TMR supplemented with native pasture and from 13 cows fed only TMR on one farm was collected separately 4 times with an interval of 15 d between collections. Two blocks (14 kg each) of cheese were made from each milk. The objective was to determine the influence of consumption of native plants in Sicilian pastures on the aroma compounds present in Ragusano cheese. Milk from cows that consumed native pasture plants produced cheeses with more odor-active compounds. In 4-mo-old cheese made from milk of pasture-fed cows, 27 odor-active compounds were identified, whereas only 13 were detected in cheese made from milk of total mixed ration-fed cows. The pasture cheeses were much more rich in odor-active aldehyde, ester, and terpenoid compounds than cheeses from cows fed only total mixed ration. A total of 8 unique aroma-active compounds (i.e., not reported in other cheeses evaluated by gas chromatography olfactory) were detected in Ragusano cheese made from milk from cows consuming native Sicilian pasture plants. These compounds were 2 aldehydes ([E,E]-2,4-octadienal and dodecanal), 2 esters (geranyl acetate and [E]-methyl jasmonate), 1 sulfur compound (methionol), and 3 terpenoid compounds (1-carvone, L(-) carvone, and citronellol). Geranyl acetate and (E)-methyl jasmonate were particularly interesting because these compounds are released from fresh plants as they are being damaged and are part of a possible plant defense mechanism against damage from insects. Most of the odor-active compounds that were unique in Ragusano cheese from pasture-fed cows appeared to be compounds created by oxidation processes in the plants that may have occurred during foraging and ingestion by the cow. Some odor-active compounds were consistently present in pasture cheeses that were not detected in the total mixed ration cheeses or in the 14 species of pasture plants analyzed. Either these compounds were present in other plants not analyzed, created in the rumen or in cheese after the pasture-plant material had been consumed, or the compounds were lost in the method of sample extraction used for the plant analysis (i.e., steam distillation) versus the solid-phase microextraction method used for the cheeses. This research has demonstrated clearly that some unique odor-active compounds found in pasture plants can be transferred to the cheese.

A mechanistic model for predicting the nutrient requirements and feed biological values for sheep.

The Cornell Net Carbohydrate and Protein System (CNCPS), a mechanistic model that predicts nutrient requirements and biological values of feeds for cattle, was modified for use with sheep. Published equations were added for predicting the energy and protein requirements of sheep, with a special emphasis on dairy sheep, whose specific needs are not considered by most sheep-feeding systems. The CNCPS for cattle equations that are used to predict the supply of nutrients from each feed were modified to include new solid and liquid ruminal passage rates for sheep, and revised equations were inserted to predict metabolic fecal N. Equations were added to predict fluxes in body energy and protein reserves from BW and condition score. When evaluated with data from seven published studies (19 treatments), for which the CNCPS for sheep predicted positive ruminal N balance, the CNCPS for sheep predicted OM digestibility, which is used to predict feed ME values, with no mean bias (1.1 g/100 g of OM; P > 0.10) and a low root mean squared prediction error (RMSPE; 3.6 g/100 g of OM). Crude protein digestibility, which is used to predict N excretion, was evaluated with eight published studies (23 treatments). The model predicted CP digestibility with no mean bias (-1.9 g/100 g of CP; P > 0.10) but with a large RMSPE (7.2 g/100 g of CP). Evaluation with a data set of published studies in which the CNCPS for sheep predicted negative ruminal N balance indicated that the model tended to underpredict OM digestibility (mean bias of -3.3 g/100 g of OM, P > 0.10; RMSPE = 6.5 g/100 g of OM; n = 12) and to overpredict CP digestibility (mean bias of 2.7 g/100 g of CP, P > 0.10; RMSPE = 12.8 g/100 g of CP; n = 7). The ability of the CNCPS for sheep to predict gains and losses in shrunk BW was evaluated using data from six studies with adult sheep (13 treatments with lactating ewes and 16 with dry ewes). It accurately predicted variations in shrunk BW when diets had positive N balance (mean bias of 5.8 g/d; P > 0.10; RMSPE of 30.0 g/d; n = 15), whereas it markedly overpredicted the variations in shrunk BW when ruminal balance was negative (mean bias of 53.4 g/d, P < 0.05; RMSPE = 84.1 g/d; n = 14). These evaluations indicated that the Cornell Net Carbohydrate and Protein System for Sheep can be used to predict energy and protein requirements, feed biological values, and BW gains and losses in adult sheep.

Contribution of native pasture to the sensory properties of Ragusano cheese.

Ragusano is a Protected Denomination of Origin cheese produced in the Hyblean area of Sicily. Sixteen samples of Ragusano cheese from two different treatments [pasture and total mixed ration (TMR)] were evaluated after 4 and 7 mo of aging. The color of the cheeses produced from milk of cows consuming fresh native pasture plants was much more yellow than cheeses from TMR fed cows (i.e., higher Hunter b value). This was due to transfer of beta-carotene and related compounds from the diet and demonstrated that compounds from native pasture plants changed the sensory characteristics of Ragusano cheese. To avoid a "halo" effect in a trained panel, quantitative descriptive analysis sensory evaluation of these cheeses for odor, taste, consistency, and mouth structure, color differences among cheeses were masked. A unique approach in sensory analysis was developed using sunglasses with lenses designed to block light at the specific wavelengths at which panelists would detect differences in color among samples. Testing was conducted every 2-wk period (15-d increments) with two tests per week using 11 trained panelists. All the panelists tasted all the products. Panelists were able to detect significant differences in the sensory characteristics of cheeses produced from milk of cows consuming native pastures versus TMR even when the color difference was masked.

Stromal support augments extended long-term ex vivo expansion of hemopoietic progenitor cells.

Current technology to numerically expand hemopoietic stem/progenitor cells (HSPC) ex vivo within 1 to 2 weeks is insufficient to warrant significant gain in reconstitution time following their transplantation. In order to more stringently test the parameters affecting HSPC expansion, we followed ex vivo cultures of CD34+-selected umbilical cord blood (UCB) HSPC for up to 10 weeks and investigated the effects of stromal support and cytokine addition. The cytokine combinations included FL + TPO, FL + TPO plus SCF and/or IL6, or SCF + IL6. To identify the HSPC in uncultured and cultured material, we determined the number of colony-forming cells (CFC), cobblestone area forming cells (CAFC), the NOD/SCID repopulating ability (SRA), and CD34+ subsets by phenotyping. The highest fold-increase obtained for CD34+ and CD34+ CD38- cell numbers was, respectively, 1197 and 30,937 for stroma-free and 4066 and 117,235 for stroma-supported cultures. In general, CFC generation increased weekly in FL + TPO containing groups up to week 5 with a 28- to 195-fold expansion whereafter the weekly CFC output stabilized. Stroma support enhanced the expansion of CAFC week 6 maximally 11-fold to 89-fold with FL + TPO + IL6. Cultures stimulated with at least FL + TPO gave an estimated 10- to 14-fold expansion of the ability of CD34+ UCB cells to multilineage engraft the BM of sublethally irradiated NOD/SCID mice at 2 weeks of stroma-free and stroma-supported cultures, while at week 5 and later the estimated SRA decreased to low or undetectable levels in all groups. Our results show that stroma and FL + TPO but also inclusion of bovine serum albumin, greatly increase the long-term generation of HSPC as measured by in vitro assays and is indispensable for long-term expansion of CD34+ CD38- CXCR4+ cells. However, the different surrogate methods to quantify the HSPC (CD34+ CD38-, CFC, CAFC week 6 and SRA) show increasing incongruency with increasing culture time, while especially the phenotypic analysis and the CFC generation greatly overestimate the CAFC and SRA expansion in 10-week cultures.

Successful short-term ex vivo expansion of NOD/SCID repopulating ability and CAFC week 6 from umbilical cord blood.

In view of the limited potential for rapid hematological recovery after transplantation of umbilical cord blood cells (UCB) in adults, we have attempted to expand CD34+ selected hemopoietic stem cells (HSC) and progenitors in 2-week cultures of whole graft pools in the presence or absence of serum and stromal layers, and with various cytokine combinations including (1) FL + TPO; (2) FL + TPO plus SCF and/or IL6; or (3) SCF + IL6. Both in the input material and cultured grafts we determined the number of colony-forming cells (CFC), cobblestone area forming cells (CAFC), the NOD/SCID repopulating ability (SRA), and CD34+ CD38- subset by phenotyping. The highest fold-increase obtained for the number of nucleated cells (nc), CD34+, CD34+ CD38 cell numbers and CFC content was, respectively, 102 +/- 76, 24 +/- 19, 190 +/- 202 and 53 +/- 37 for stroma-free and 315 +/- 110, 25 +/- 3, 346 +/- 410 and 53 +/- 43 for stroma-supported cultures. CAFC week type 6 was maximally 11-fold expanded both under stroma-free and stroma-supported conditions. The FBMD-1 stromal cells supported a modest expansion of CD34+ CD38- cells (27 +/- 18-fold) and nc (6 +/- 4-fold), while a loss of CFC and CAFC subsets was observed. The stromal cells synergized with FL + TPO to give the highest expansion of hemopoietic progenitors. Stromal support could be fully replaced by complementing the FL + TPO stimulated cultures with SCF + IL6. FL + TPO were required and sufficient to give a 10- to 20-fold expansion of the ability of CD34+ UCB cells in 2-week cultures to engraft the BM of NOD/SCID mice. Stromal support, or complementation of the medium with SCF + IL6, did not significantly improve the in vivo engraftment potential. If the SRA and CAFC week 6 assays are accepted as tentative estimates of in vivo engrafting stem cells in humans, our findings may assist in the preparation of UCB grafts to meet the requirements for improved repopulation in the clinical setting.

Activation of protein kinase C by oxytocin-related conopressin underlies pacemaker current in Lymnaea central neurons.

The vasopressin/oxytocin-related neuropeptide Lys-conopressin activates two pacemaker currents in central neurons of the mollusk Lymnaea stagnalis. A high-voltage-activated current (I-HVA) is activated at potentials greater than -40 mV and resembles pacemaker currents found in many molluscan neurons. A low-voltage-activated current (I-LVA) activates throughout the range of -90 to 0 mV. Based on sequence homologies, Lymnaea conopressin receptors are thought to couple to Q-type G proteins and protein kinase C (PKC). Alternatively, agonist-induced pacemaker currents in molluscan neurons have traditionally been attributed to cAMP-dependent protein kinase (PKA) activation. Accordingly, this study aimed at resolving possible involvement of cAMP/PKA and diacylglycerol/PKC in the conopressin response. Injection of cAMP into anterior lobe neurons induced a slow inward current with a voltage dependence resembling that of I(LVA) (and not I(HVA)). However, lack of effect of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine and the absence of cross-desensitization between cAMP and conopressin suggest that neither current is dependent on intracellular cAMP. The PKC-activating phorbol ester 12-O-tetradecanoylphorbol 13-acetate (but not inactive phorbol 12-myristate 13-acetate) mimicked activation of I(HVA), but not I(LVA), and occluded subsequent responses to conopressin. Activation of I(HVA) was blocked by general protein kinase inhibitors and the PKC-inhibitor GF-109203X. Modulation of the calcium buffering capacity of the pipette medium did not affect the conopressin response, suggesting that calcium dynamics are not of major importance. We conclude that conopressin activates the ion channels carrying I(LVA) and I(HVA) through different second-messenger cascades and that PKC-dependent phosphorylation underlies activation of I(HVA).

Assessment of the dairy production needs of cattle owners in southeastern Sicily.

This study was undertaken to investigate research and outreach priorities for Progetto Ibleo (Project Ibleo), a center created in 1990 with tripartite government funding to serve dairy producers in the Hyblean region of Sicily. Data comprised values for production and composition of milk from 1984 to 1989 from 35 herds of Modicana cows on a system based on pasture and that from 69 input-intensive herds of Holstein cows, associated lactation and reproduction measures, and yield and composition of forages from 4 of these farms in 1988. Season had a large effect on the neutral detergent fiber and crude protein composition of forages, production and composition of milk, and predicted yield of fresh Ragusano cheese manufactured from the milk of these cows. The poorest forage quality and the poorest cow performance were observed in summer and fall months (May to October). Lactation curves that were flat, without a discernible peak, or convex were observed for both systems, especially for cows calving in spring and in the dry summer seasons (March to July). These abnormalities, signifying substantial sacrifices in production potential, probably had a complex etiology that stemmed from low nutrient intake and high neutral detergent fiber and low crude protein composition of the grazed and preserved forages. Research and outreach priorities to support the Hyblean dairy industry should include chemical evaluation of forages and other feedstuffs, low moisture ensiling of high quality winter forages, better formulation of diets that are dense with nutrients, and the shifting of calving patterns to better exploit high quality winter forages.

Predicting forage indigestible NDF from lignin concentration.

We used chemical composition and in vitro digestibility data from temperate and tropical forages to develop relationships between indices of lignification and forage indigestible NDF. Neutral detergent fiber indigestibility increased nonlinearly as the lignin concentration of the NDF increased. Differences in estimated indigestible NDF using equations developed for a specific forage class (C3 and C4 grasses and legumes) were small and are probably not biologically significant when compared to those estimated from a common equation. Selected equations were compared with the Cornell Net Carbohydrate and Protein System (CNCPS) for the prediction of ADG. The linear equation (2.4 times NDF lignin content) used by the CNCPS and the Beef NRC had some of the largest errors due to mean bias. A log-log model [4.37 x (lignin/NDF)(.84)] provided the best combination of low total prediction error, low mean bias, and minimal error due to regression bias when permanganate lignin was used. A similar equation based on sulfuric acid lignin [6.17 x (lignin/NDF)(.77)] also met the above criteria. These equations then were evaluated with the CNCPS model against animal growth data from diets ranging in forage quality. Regardless of the equation used for predicting unavailable fiber, the CNCPS underpredicted daily gain, with mean biases ranging from -.10 to -.22 kg/d. Regression bias ranged from .13 to .14 kg/d and the coefficients differed from unity (P = .0001). The new equations gave numerically lower energy allowable ADG by steers compared to the linear equation currently used by the CNCPS model. The estimates were lower due to a higher predicted indigestible NDF, which resulted in a lower estimated forage energy value.

Conopressin affects excitability, firing, and action potential shape through stimulation of transient and persistent inward currents in mulluscan neurons.

The molluscan vasopressin/oxytocin-related neuropeptide conopressin activates two persistent inward currents in neurons from the anterior lobe of the right cerebral ganglion of Lymnaea stagnalis that are involved in the control of male copulatory behavior. The low-voltage-activated (LVA) current is activated at a wide range of membrane potentials, its amplitude being only weakly voltage dependent. The high-voltage-activated (HVA) current is activated at potentials positive to -40 mV only and shows a steep voltage dependence. Occurrence of both currents varies from cell to cell, some expressing both and others only the HVA current. In most neurons that have the LVA current, a conopressin-independent persistent inward current (INSR) is found that resembles the HVA current in its voltage dependence. The functional importance of the LVA and HVA currents was studied under current-clamp conditions in isolated anterior lobe neurons. In cells exhibiting both current types, the effect of activation of the LVA current alone was investigated as follows: previously recorded LVA current profiles were injected into the neurons, and the effects were compared with responses induced by conopressin. Both treatments resulted in a strong depolarization and firing activity. No differences in firing frequency and burst duration were observed, indicating that activation of the LVA current is sufficient to evoke bursts. In cells exhibiting only the HVA current, the effect of conopressin on the response to a depolarizing stimulus was tested. Conopressin reversibly increased the number of action potentials generated by the stimulus, suggesting that the HVA current enhances excitability and counteracts accommodation. Conopressin enhanced action potential broadening during depolarizing stimuli in many neurons. Voltage-clamp experiments performed under ion-selective conditions revealed the presence of transient sodium and calcium currents. Using the action potential clamp technique, it was shown that both currents contribute to the action potential. The calcium current, which is activated mainly during the repolarizing phase of the action potential, is augmented by conopressin. Thus conopressin may directly modulate the shape of the action potential. In summary, conopressin may act simultaneously on multiple inward currents in anterior lobe neurons of Lymnaea to affect firing activity, excitability, and action potential shape.

Vasopressin/oxytocin-related conopressin induces two separate pacemaker currents in an identified central neuron of Lymnaea stagnalis.

The molluscan vasopressin/oxytocin analogue Lys-conopressin excites neurons in the anterior lobe of the right cerebral ganglion of the snail Lymnaea stagnalis. Persistent inward currents that underlie the excitatory response were studied with the use of voltage-ramp protocols in the identified neuron RCB1 and other anterior lobe neurons. Under whole cell voltage-clamp conditions, two types of conopressin-activated current could be distinguished on the basis of their voltage dependence: 1) a pacemaker-like current that was activated at potentials above -40 mV (high-voltage-activated current, I(HVA)) and 2) an inward current that was activated at all potentials between -90 and +10 mV (low-voltage-activated current, I(LVA)). Ion substitution experiments indicate that sodium is the main charge carrier for I(HVA) and I(LVA). Both currents are differentially affected by cadmium. I(HVA) and I(LVA) differ in dose dependence, with median effective concentration values of 7.7 x 10(-8) M and 2.2 x 10(-7) M, respectively. Vasopressin and oxytocin act as weak agonists for the conopressin responses. The kinetics of desensitization and washout of I(HVA) and I(LVA) are different. The HVA response shows little desensitization, whereas the LVA response desensitizes within minutes (time constant 80 +/- 28 s, mean +/- SD). The time constant of washout on removal of conopressin is 159 +/- 63 s for I(HVA) and 36 +/- 13 s for I(LVA). These results suggest that two distinct conopressin receptors are involved in the activation of both currents. The conopressin-activated currents induce or enhance a region of negative slope resistance in the steady-state current-voltage relation. They differ from a third persistent inward current that is carried by calcium and completely blocked by cadmium. The presumed functional roles of these currents, possibly including autoregulation, are discussed.

Studies on the effects of selenium on rumen microbial fermentation in vitro.

The effects of selenium (Se) on ruminant microbial fermentation were investigated in vitro using rumen microflora collected from a rumen-fistulated dairy cow. First, the effects of L-selenomethionine (SeMet; at 0.2 or 2 ppm Se) in the presence or absence of wheat bran (WB, 500 mg per incubation flask) were evaluated. Second, the effects of several forms of Se (elemental Se: 50 ppm Se; sodium selenite: 2 ppm Se; SeMet: 2 ppm Se) were compared. Results showed that the amounts of short-chain fatty acids (SCFAs) tended to be increased by SeMet treatment, whereas SeMet in the presence of WB transiently suppressed fermentation. The addition of SeMet tended to increase the production of acetate while reducing the production of butyrate with the without WB supplementation. Among the different Se compounds tested, the amounts of SCFAs were greater with SeMet treatment, which yielded a higher proportion of acetate compared to other treatments. Selenite did not influence the total SCFAs concentrations; however, it increased the relative proportion of butyrate at the expense of acetate. Elemental Se did not significantly affect fermentation. Higher bacterial Se concentrations were observed for selenite than for SeMet. It was concluded that Se supplementation can influence rumen microbial fermentation and that Se compounds differ in this regard.

Prediction of ruminal volatile fatty acids and pH within the net carbohydrate and protein system.

A steady-state model of the production, absorption, passage, and concentration of ruminal VFA and pH is developed from published literature data and is structured to use the feed descriptions and inputs from the net carbohydrate and protein system. Included are the effects of pH on growth rate and yield of structural and non-structural carbohydrate-fermenting bacteria; production of acetate, propionate, butyrate, lactate, and methane; conversion of lactate to VFA; ruminal absorption of acids; and prediction of ruminal pH from dietary measures and from ruminal buffering and acidity. The root mean square error of predicted total VFA concentration was 12 mM. Individual VFA fractions were inadequately predicted. In a review of literature data, effective NDF (eNDF) provided a better correlation with ruminal pH than forage or NDF. Digestion rate of NDF remained at normal levels above pH 6.2, which corresponds to a minimum eNDF of 20% of dietary DM. Further research is needed to determine the individual VFA produced from carbohydrate fractions at various pH, the appropriateness of partitioning the starch and pectin carbohydrate pool into slowly and rapidly degraded fractions, and the effect on microbial yield, total tract digestibility, and predicted energy values of feeds.

Comparison of Landim and Africander cattle in southern Mozambique: I. Body weights and growth.

The growth performance of Landim and Africander breeds was compared using data collected from 1968 to 1981 at the Chobela Research Station in Mozambique. Animals from both breeds were managed together in groups by age and sex, except when separated for breeding. Growth traits were body weights at birth, weaning at 7 mo, 18 mo, and first calving, and pre- and postweaning daily growth rates. These traits were analyzed using a mixed-effects least squares model containing breed, year-season of birth, sex, the nested effect of parity within breed, a linear regression on dam's age, and the random effect of sire within breed. Africander calves were 16, 9, and 7% heavier (P < .01) than Landim calves at birth, weaning, and 18 mo (18 +/- 6 kg heavier than the 237-kg Landim average). However, there was no detectable difference for age-adjusted weight at first calving and postweaning daily growth rate. Diminishing weight and growth differences with advancing age may indicate adaptation by the Landim to the prevailing environmental limitations in southern Mozambique, especially through younger ages at puberty and at first calving.

Comparison of Landim and Africander cattle in southern Mozambique: II. Female fertility, reproduction, and beef offtake.

Fertility and reproductive performance of Landim and Africander females were compared using data collected from 1968 to 1981 at the Chobela Research Station in Mozambique. Breeds were managed together and grouped by age and sex, except when separated for breeding. Traits were relative fertility (probability of fertile females calving from the first breeding season), age at first calving, first calving interval, and subsequent calving intervals. calving rates were tested by x2 procedures with equal expected frequencies in each subclass. The statistical model included breed, the random effect of sire within breed, year-season of birth or calving, and calving group within breed. Landim survivors were more fertile (P < .05) than the Africander ones throughout their recorded lifetimes. Landim females were 1.32 +/- .21 mo (or 3%) younger at first calving and had a 48 +/- 12 d (or 11%) shorter interval between first and second calving than the Africander average of 473 d. When reproductive and growth information were combined to compute an annual index of beef offtake expressed as 18-mo calf yield per unit of dam's weight at first calving, Landim cows annually yielded 30% more calf weight (P < .001) than Africander cows per kilogram of their own body maintenance despite lighter body weights at 18 mo. Superior fertility of Landim females led to greater beef offtake from higher calving rates. Greater fertility and relatively less feed to maintain the reproducing herd are probable mechanisms for a population to adapt to nutrient-limiting environments such as the one in southern Mozambique.

A novel G protein-coupled receptor mediating both vasopressin- and oxytocin-like functions of Lys-conopressin in Lymnaea stagnalis.

We have cloned a receptor, named LSCPR, for vasopressin-related Lys-conopressin in Lymnaea stagnalis. Lys-conopressin evokes Ca(2+)-dependent Cl- currents in Xenopus oocytes injected with LSCPR cRNA. Expression of LSCPR mRNA was detected in central neurons and peripheral muscles associated with reproduction. Upon application of Lys-conopressin, both neurons and muscle cells depolarize owing to an enhancement of voltage-dependent Ca2+ currents and start firing action potentials. Some neurons coexpress LSCPR and Lys-conopressin, suggesting an autotransmitter-like function for this peptide. Lys-conopressin also induces a depolarizing response in LSCPR-expressing neuroendocrine cells that control carbohydrate metabolism. Thus, in addition to oxytocin-like reproductive functions, LSCPR mediates vasopressin-like metabolic functions of Lys-conopressin as well.

The local anesthetic n-butyl-p-aminobenzoate selectively affects inactivation of fast sodium currents in cultured rat sensory neurons.

BACKGROUND: Aqueous suspensions of the local anesthetic n-butyl-p-aminobenzoate (BAB), epidurally applied in terminal cancer patients, resulted in a sensory blockade, lasting up to several months. To investigate the mechanism of action on the cellular level, the effect of 100 microM BAB on Na+ action potentials and on Na+ currents in dorsal root ganglion neurons from neonatal rats was studied. METHODS: Small neurons grown in cell culture were selected for patch-clamp measurements. Both Na+ action potentials, evoked by current pulses of increasing amplitude (current clamp) and Na+ currents, activated at different membrane potentials (voltage clamp), were investigated in the absence and presence of 100 microM BAB. The local anesthetic was applied by external perfusion for 2 or 10 min. RESULTS: In the presence of 100 microM BAB, either the firing threshold was raised or the action potential was abolished. The maximal peak conductances, underlying the fast sodium current INa,F and the slow sodium current INa,5, were not changed. However, the inactivation of INa,F was increased by BAB. The sigmoid inactivation curve shifted 12 mV toward hyperpolarizing membrane voltages, whereas no changes were found for the inactivation of the slow Na+ current. Only at short exposure times of 2 min, the effects of BAB could be reversed during a 10-min wash-out. CONCLUSIONS: BAB dramatically increased the firing threshold, and in part of the sensory neurons, it blocked the action potential. The inactivation of the fast Na+ channels, but not of the slow Na+ channels, was increased by BAB. Thus, the block of fast Na+ channels by BAB may contribute to epidural analgesia. At exposure times of 10 min, the effect of BAB was not reversible. This probably originates from its high lipid-solubility, which may be an important factor in determining the duration of the block in vivo.

A common stem cell for murine cortical and medullary thymic epithelial cells?

We have addressed the question whether the epithelial stroma in the thymus is derived from a common stem cell or whether cortical and medullary epithelial cells are derived from different embryonic stem cells emerging, for example, from endoderm and ectoderm. By the use of rapidly expanding cultures of thymic epithelial cells (TEC) from 14 to 16 day-old murine fetuses and by specific antibodies against cortical and medullary epithelium, respectively, we were able to demonstrate a small subpopulation of double-labeled TEC in the cultures. These cells were not present in TEC cultures initiated from thymuses of neonatal mice. Double-labeled TEC were also found in tissue sections from fetal thymuses. These findings may indicate that TEC populations of the cortex and the medulla are derived from a common stem cell, with potential for differentiation toward both cortical and medullary TEC.

Transferrin receptor expression as a marker of immature cycling thymocytes in the mouse.

Dividing cells require iron and, therefore, express the transferrin receptor (CD71) on the cell surface to enable internalization of transferrin-bound iron. Since early T cell development is marked by intense proliferation, we questioned whether CD71 might serve as a marker of immature T cells. Therefore, we analyzed the expression of CD71 on fetal, neonatal, and adult thymocytes in correlation with cell size, cell cycle status, and expression of CD3, CD4, CD8, alpha beta TcR, and gamma delta TcR. Phenotypic analysis showed that only the large, immature CD4-8-3-, CD4-8+3-, and CD4+8+3- cells in fetal, neonatal, and adult thymus expressed CD71. In addition, DNA analysis showed that all CD71+ large adult thymocytes were cycling. Downregulation of CD71 occurs when proliferation ceases, i.e., within the CD4+8+3- thymocyte subpopulation. The gradual changes in size and CD71 expression suggest a sequential development within this CD4+8+3- subpopulation from large CD71+ via small CD71+/- to small CD71- cells. As a consequence, CD71 expression is downregulated, in adult T cell development as well as in ontogeny, before the alpha beta TcR appears on the cell surface of the thymocyte. Together, our findings show that CD71 is a marker of immature, proliferating T cells.