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Yield and composition of milk from lactating mares: effect of lactation stage and individual differences.

The yields and composition of milk from nursing mares were studied during the first two months of lactation in 11 mares of heavy breeds (784 kg). Daily yield increased from 21.7 to 24.6 kg between weeks 1 and 8 of lactation. Fat, protein, gross energy and Ca concentrations significantly decreased when lactose content increased during this period. Individual variations were higher for yield than for composition. Casein, whey protein and non-protein N (56, 34 and 10% of crude protein, respectively) and amino acid composition did not vary between weeks 1 and 8 of lactation.

Amino Acids↗

Functional atrophy of brown adipose tissue during lactation in mice. Effects of lactation and weaning on mitochondrial GDP binding and uncoupling protein.

The thermogenic activity and capacity of brown adipose tissue were determined in mice during lactation and after weaning. There was a marked fall during lactation in the mitochondrial content of the tissue, and in GDP binding and the specific mitochondrial concentration of uncoupling protein. The lactation-induced functional atrophy of brown adipose tissue was fully reversible after weaning; mitochondrial content and the mitochondrial concentration of uncoupling protein were both restored, although GDP binding was not normalized.

Adipose Tissue, Brown↗

Relationship between ovarian delta 5-3 beta-hydroxysteroid dehydrogenase activity and implantation in non-lactating and lactating mongolian gerbils (Meriones unguiculatus).

The distribution and intensity of delta 5-3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activity was examined histochemically in ovaries from Mongolian gerbils during pregnancy and undergoing lactational delay. In all of the ovaries examined the corpora lutea showed the strongest level of activity, but like the interstitium and granulosa, there was no relationship between the enzyme pattern and embryonic development. Thecal 3 beta-HSD activity was only observed immediately before, and after implantation on day 8 p.c. in pregnancies without lactation, and was totally absent throughout lactational delay, appearing for the first time after day 15 in association with implantation. The absence of this key enzyme in the theca interna throughout the pre-implantation period means that the biosynthesis of androstenedione and progesterone cannot occur, thereby precluding the production of oestrogen, an essential steroid for implantation in this species.

3-Hydroxysteroid Dehydrogenases↗

Prediction of lactate threshold and fixed blood lactate concentrations from 3200-m running performance in male runners.

To determine the effectiveness of a 3200-m time trial for predicting VO2 and running velocity at lactate threshold (LT), fixed blood lactate concentrations of 2.0, 2.5, and 4.0 mM, and peak, 42 male runners (means age = 31.1 +/- 8.3 years; means ht = 176.8 +/- 6.6 cm; means wt = 70.4 +/- 10.0 kg) completed a VO2 peak/LT test and a 3200-m time trial. The continuous treadmill protocol started at 0% grade 150 m/min and increased 10 m/min every 3 min until exhaustion. Velocity at LT, 2.0, 2.5, and 4.0 mM was determined from individual velocity blood lactate relationships, and VO2 values were determined from individual plots ov VO2 vs velocity. VO2 peak and velocity peak were chosen as the highest values observed. Oxygen uptake at LT, 2.0, 2.5, 4.0 mM, and peak was 52.51, 56.61, 58.31, 61.70, and 64.21 ml/kg.min-1, respectively, while the velocities associated with LT, 2.0, 2.5, 4.0 mM, and peak were 235.5, 251.5, 259.8, 273.5, and 285.5 m/min, respectively. During the 3200-m time trial (means time = 11.28 +/- 0.96 min), 400-m split times and cumulative times were recorded. Twenty-nine subjects were randomly assigned to a validation sample and the remaining subjects were used for cross-validation purposes. Regression analysis revealed that a 3200-m time trial was a good predictor of both VO2 (ml/kg.min-1) and velocity (m/min) at LT, 2.0, 2.5, 4.0 mM, and peak.

Adult↗

Effects of pedal frequency on VO2 and work output at lactate threshold (LT), fixed blood lactate concentrations of 2 mM and 4 mM, and max in competitive cyclists.

To determine the effects of differing pedal frequencies on VO2 and work output values at the lactate threshold (LT), fixed blood lactate concentrations of 2 mM and 4 mM (2 mM, 4 mM), and at max, nine male competitive road racing cyclists (USCF category I or II) completed three VO2 max tests; on a Monark bicycle ergometer, at pedal frequencies of 60, 90, and 120 rpm. Each stage was 3 min in duration, starting at 0 kgm/min with subsequent stages increased by either 180 kgm/min (60 and 120 rpm) or 178 kgm/min (90 rpm). Blood samples were taken during the last 30 s of each stage. VO2 and work output at LT, 2 mM, and 4 mM were determined from individual blood lactate-work rate and VO2-work rate relationships. VO2 max and maximal work output were chosen as the peak values observed during the VO2 max tests. Results indicated that work output at LT, 2 mM, and 4 mM was affected by choice of pedal frequency (1278, 1140, 999 kgm/min at LT; 1533, 1450, 1182 kgm/min at 2 mM; 1780, 1703, 1487 kgm/min at 4 mM; for 60, 90, and 120 rpm, respectively, P less than 0.05). Max work output at 60 and 90 rpm was significantly greater (P less than 0.05) than at 120 rpm (2035, 2053, 1879 kgm/min for 60, 90, and 120 rpm, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Effect of calcium supplements and stage of lactation on the calcium absorption efficiency of lactating women accustomed to low calcium intakes.

The effect of calcium intake on the calcium absorption efficiency from 100 mL cow milk was measured in lactating Gambian mothers habituated to a low-calcium diet [mean intake 7.08 mmol (283 mg)/d], and compared with UK lactating mothers consuming high-calcium diets [mean intake 29.2 mmol (1168 mg)/d] by using a double stable-isotope technique (oral 44Ca and intravenous 42Ca). In a double-blind trial starting 9 d postpartum, Gambian mothers were given a calcium supplement [17.85 mmol (714 mg)/d] or placebo for 12 mo. At 3 and 12 mo postpartum, mean (+/- SEM) calcium absorption from isotopically enriched milk was 52.3 +/- 3.1% (n = 25) and 47.2 +/- 4.8% (n = 24) in the unsupplemented Gambian mothers and 48.8 +/- 2.8% (n = 28) and 42.9 +/- 3.7% (n = 24) in the supplemented mothers, respectively. There was no effect of supplementation or stage of lactation on the efficiency of calcium absorption. At 3 mo postpartum the UK mothers absorbed 32.2 +/- 3.8% of the isotopically enriched calcium added to milk, which was significantly less than that of the Gambian mothers (P < 0.01).

Absorption↗

Loss of tissues in female rats subjected to food restriction during lactation or during both gestation and lactation.

The effects of food restriction (50% of ad libitum-fed controls) on maternal tissues during lactation (LR) or during both gestation and lactation (GLR) were examined in rats. Skeletal muscle protein, RNA and fiber diameter were reduced in response to the LR and GLR treatments, but muscle DNA and fiber number were unchanged. Mean body weights of the pregnant (GLR) and nonpregnant female rats were not significantly different after the first 3 wk of food restriction (after delivery in pregnant GLR rats). Compared to the controls, pups of the GLR females had a significantly smaller mean birth weight. The LR females lost weight and catabolized fat pads, organs, and skeletal muscles while nonlactating females gained weight when given a similar amount of food. These results indicated that the demands of gestation did not significantly add to the loss of maternal body weight due to the restriction of food intake. However, the demands of lactation caused the food-restricted females to catabolize significant amounts of maternal tissues.

Adipose Tissue↗

Fructose 1,6-bisphosphate-dependent L-lactate dehydrogenase from Thermus aquaticus YT-1, an extreme thermophile: activation by citrate and modification reagents and comparison with Thermus caldophilus GK24 L-lactate dehydrogenase.

Heat-stable fructose 1,6-bisphosphate-dependent L-lactate dehydrogenase [EC 1.1.1.27] was purified from an extremely thermophilic bacterium, Thermus aquaticus YT-1. The amino acid composition and NH2-terminal 34 amino acid sequence of the enzyme were determined. Its NH2-terminal sequence shows high homology with those of Thermus caldophilus GK24 (82% identity) and some other bacterial L-lactate dehydrogenases (44-53% identity), indicating the close phylogenic relationship of the two Thermus species. At the same time, the two Thermus L-lactate dehydrogenases were found not to be identical not only chemically but also kinetically and immunologically. Citrate activated the T. aquaticus enzyme in the weak acidic pH region, while fructose 1,6-bisphosphate did in both acidic and neutral pH regions. The maximum activity obtained with citrate at pH 5.0 was about 2.5 times higher than that in the presence of fructose 1,6-bisphosphate at pH 6.7. The enzymes modified with 2,3-butanedione, acetic anhydride and diethyl pyrocarbonate in the presence of both NADH and oxamate were desensitized to fructose 1,6-bisphosphate, and the modified enzymes were active even in the absence of fructose 1,6-bisphosphate. All of the modified enzymes examined were still activated by citrate similarly to the native enzyme. These results suggest that the mechanism of activation by citrate is different from that by fructose 1,6-bisphosphate, and that the citrate-binding site is different from the fructose 1,6-bisphosphate-binding site.

Amino Acids↗

Immunological study of lactate dehydrogenase from Streptococcus mutans and evidence of common antigenic domains with lactate dehydrogenases from lactic bacteria.

Rabbit polyclonal antibodies directed against purified Streptococcus mutans L-(+)-lactate dehydrogenase reacted with the purified enzyme, giving a marked deviation of its kinetic parameters. The enzyme affinity for pyruvate or NADH decreased in the presence of antibody, the affinity for fructose 1,6-diphosphate (FDP) appeared to be slightly affected, and the cooperativity of the ligand binding was lowered. A partial protective effect was observed when the enzyme was preincubated with FDP prior to the antibody adjunction. An enzyme-linked immunosorbent assay allowed detection of a 30% decrease in enzyme-antibody fixation when FDP was added. The protective effect observed with FDP could be correlated with a conformational change induced by the activator. A decrease of antibody binding in the presence of FDP was also obtained with S. sanguis, Actinomyces viscosus, and Lactobacillus casei lactate dehydrogenases, which reflects a similar mechanism of activation among lactic bacteria. NADH did not offer any protection against antibody inhibition or fixation, and the coenzyme affinity decrease could be attributed to an indirect mechanism. On the contrary, pyruvate and the immunoglobulins apparently could compete for specific binding sites. A decrease of antibody binding was also obtained with three heterologous lactic bacterial lactate dehydrogenases, indicating a conservation of antigenic determinants implicated in the substrate binding.

Actinomyces↗

Effect of lactate concentration and metabolic rate on net lactate uptake by canine skeletal muscle.

This study addressed two questions: 1) Does net lactate uptake (L) by muscle approach a saturation limit with increasing blood lactate concentration ([La])? 2) Is the muscle net L response to increasing blood [La] affected by metabolic rate (VO2)? The gastrocnemius plantaris muscle group (GP) was isolated in situ in 20 anesthetized dogs. In three series of experiments, a lactate-lactic acid solution was infused into the arterial inflow of the GP to produce five different plasma [La] values: approximately 3, 9, 16, 22, and 30 mM, each of them maintained for 30 min. In one series, the GP remained at rest, whereas in the second series it contracted at 1 Hz and in the third series at 4 Hz. VO2 averaged approximately 3, 43, and 100 ml.kg-1.min-1 at rest and at 1 and 4 Hz, respectively. Within each of the three metabolic rates, increasing plasma [La] resulted in an increase in net L, which was well described (R > 0.98) by exponential equations. These equations predicted net L asymptotic values of 0.80, 0.72, and 1.09 mmol.kg-1.min-1 for rest and for 1 and 4 Hz, respectively. The corresponding plasma [La]s for half-maximal net L from the exponential equations were 16, 10, and 12 mM. Glucose uptake, pyruvate uptake/output, and alanine output by the muscles were not affected by the increasing [La] (and concomitant increases in net L) at any of the metabolic rates. Neither net glycogen synthesis nor depletion was changed by increasing [La].(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Structure and function of L-lactate dehydrogenases from thermophilic and mesophilic bacteria. I) Isolation and characterization of lactate dehydrogenases from thermophilic and mesophilic bacilli.

Lactate dehydrogenases from thermophilic bacilli (Bacillus stearothermophilus, Bacillus caldotenax) and from mesophilic bacilli (Bacillus X1, Bacillus subtilis) have been isolated by a two-step purification procedure. Only one type (LDH-P4) composed of four identical subunits (Mr 34 000 or 36 000) was found in each bacillus. The tetrameric enzymes were characterized with respect to thermostability, pH and temperature dependence of the pyruvate reduction and the L-lactate oxidation, substrate specificity, saturation kinetics (Km values of pyruvate, lactate, NAD, NADH), pyruvate and oxamate inhibition, and activation by fructose bisphosphate. The thermophilic and mesophilic enzymes differ characteristically in these parameters. Preliminary structural data (amino acid composition, comparative N-terminal sequence analysis) show the expected close phylogenetic relationship (high degree of sequence homology), but also typical differences between thermophilic and mesophilic dehydrogenases, a suitable basis for further comparative studies.

Bacillus↗

Structure and function of L-lactate dehydrogenases from thermophilic and mesophilic bacteria, IV. The primary structure of the mesophilic lactate dehydrogenase from Bacillus subtilis.

The complete amino-acid sequence of lactate dehydrogenase from the mesophilic Bacillus subtilis (B. X1) was determined. Approximately 70% of the sequence was obtained by sequence analysis of intact protein (N-terminal sequence) and of four CNBr fragments (CNBr3, CNBr4, CNBr5 and CNBr6). Sequences overlapping the CNBr fragments were determined from polypeptide fragments obtained by cleavage using o-iodosobenzoic acid (cleavage at Trp) or clostripain (cleavage at Arg). The C-terminal amino-acid residue (Asn) was detected by carboxypeptidase Y-degradation. Lactate dehydrogenase from B. subtilis shows a 69% sequence homology to that from the thermophilic strain B. stearothermophilus, and a 34% sequence homology to those from higher organism. The homology of these enzymes is particularly high at the active site regions (the coenzyme and substrate binding sites). The relatively high sequence conservation of the lactate dehydrogenases from B. subtilis and B. stearothermophilus (and from other bacilli) allows a structural comparison of this temperature variants.

Amino Acid Sequence↗

Structure and function of L-lactate dehydrogenases from thermophilic and mesophilic bacteria, V. The complete amino-acid sequence of the mesophilic L-lactate dehydrogenase from Bacillus megaterium.

The complete amino-acid sequence of L-lactate-dehydrogenase from the mesophilic Bacillus megaterium was determined. 92% of the 318 amino acids were established by sequence analysis of the N-terminus, of four CNBr fragments and of one fragment obtained by cleavage with BNPS-skatole. The primary structure was completed by sequencing overlapping fragments obtained by further cleavage of suitable CNBr fragments and BNPS fragments with either trypsin, endoproteinase Lys-C, o-iodosobenzoic acid or hydroxylamine. The C-terminal amino acids were determined by degradation with carboxypeptidase A. The sequence homology between lactate dehydrogenases from B. megaterium and those from other Bacilli is 59-61% and 35-37% to those from higher organisms. The high sequence homology among lactate dehydrogenases from Bacilli, adapted to different temperatures, allows comparative studies of the structural basis of protein thermostability.

Amino Acid Sequence↗

Energy balances of lactating sows in relation to feeding level and stage of lactation.

Twelve crossbred sows were used in an energy balance study to estimate energetic efficiency of milk production from feed. Balances were made from 7 to 14 d and from 18 to 25 d of lactation. Two feeding levels were applied (high and low). The low level (L) was meant to supply energy slightly above maintenance, with energy needed for milk to be derived from body reserves. The high level (H) was meant to supply sufficient energy for maintenance and for milk production. The low-level animals received 2.5 to 2.6 kg of feed/d and the high level animals 4.8 to 6 kg/d. The loss of weight of sows during lactation depended more on feeding level than on stage of lactation. A high level of feeding to sows resulted in heavier piglets compared with the low feeding level (significant after 10 d). At 24 d of age piglets with the high-feeding-level sows weighed 7.5 kg and those with the low-level sows, 5.7 kg. At the high level, animals excreted 1,200 to 2,800 kcal more milk energy per day than the low-level animals. Energy for milk from feed was produced with an efficiency of 67 to 69% with a maintenance requirement of 112 to 125 kcal metabolizable energy (ME) X W-.75 X d-1. Efficiency of milk production from feed was calculated as 62% and the maintenance requirement was 68 kcal. In this calculation, milk was corrected toward zero energy balance. Another way of calculating this efficiency after correcting feed toward zero energy balance resulted in estimates of 68% for efficiency and of 88 kcal ME X W-.75 X d-1 for maintenance requirement. From these data it was derived that, for each piglet, the sow needed to receive .5 to .6 kg of extra feed (ME content 3,000 kcal/kg) per day to cover milk production. Level of metabolic rate for nursing piglets was estimated as 97 kcal ME X W-.75 X d-1 for maintenance and, in addition, .195 kcal/kcal extra of milk intake above maintenance.

Animal Feed↗

Effects of adding fat to the sow lactation diet on lactation and rebreeding performance.

One-hundred-three multiparous sows were randomly assigned to one of two lactation diets containing either no supplemental animal fat (C) or 10% added fat (F) during two seasons, summer (S) and winter (W), in a 2 X 2 factorial arrangement of treatments. Sows were placed on their respective dietary treatments 1 wk prior to farrowing and were fed these diets ad libitum throughout the 28-d lactation period. Weekly feed intake and total feed intake were not affected by diet or season, while weekly metabolizable energy (ME) intake tended to be higher during week 1 and 3, and was higher (P less than .04) during wk 2 for sows fed diet F. Sow weight loss from farrowing to 21 d of lactation and to weaning (28 d) were unaffected by diet or season. Average pig birth weight was .15 kg higher (P less than .01) for pigs born during S compared with those born in W. Sows receiving diet F had heavier litters at 21 d (P less than .01) and heavier average pig 21-d weights (P less than .01). This was primarily due to the 13.1% increase (P less than .04) in estimated milk yield and the higher fat concentration (P less than .001) of milk consumed by the pigs nursing sows fed diet F. Interval between weaning and rebreeding was shortened by 5.9 d (P less than .01) for sows during W than during S, and tended to be lower for sows fed diet F (7.3 d) compared with that of sows fed diet C (9.7 d).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Lactate determination with the lactate analyser LA 640: a critical study.

Using the new method of sample preparation developed by the manufacturer in 1979, the lactate analyser LA 640 now provides reliable measurements of blood lactic acid from blood micro-samples of 20 microliters within 2 min. Reproducibility is comparable to that obtained by spectrophotometric assay with +/- 0.2 mmol . l-1 for 95% confidence limits. High accuracy was also obtained with 95% linked confidence limits within +/- 0.19 mmol . l-1 and +/- 0.46 mmol . l-1 for blood lactate concentrations of from 2-4 and 7-9 mmol . l-1 respectively. Due to a lactate concentration increase of 0.5 mmol . l-1 . h-1 in whole blood kept at 20 degrees C, the blood must be immediately diluted in the buffer haemolysing solution after withdrawal. This diluted sample can then be stored at room temperature without alteration, provided the concentration in the dilution is lower than 0.8 mmol . l-1. For serial successive measurements, a new sample can be tested before the electrode current reaches its background value.

Blood Chemical Analysis↗

Effects of amounts and degradability of dietary protein on lactation, nitrogen utilization, and excretion in early lactation Holstein cows.

Five treatment diets varying in crude protein (CP) and rumen undegradable protein (RUP) were calculated to supply a postruminal lysine to methionine ratio of about 3:1. Diets were fed as a total mixed ration to 65 Holstein cows that were either primiparous (n = 28) or multiparous (n = 37) from 21 to 120 d in milk to determine effects on lactation and nitrogen utilization. Crude protein % and calculated RUP (% of CP) of diets [on a dry matter (DM) basis] were: 1) 19.4, 40 (HPMU), 2) 16.5, 34 (LPLU), 3) 16.8, 40 (LPMU), 4) 16.8, 46 (LPHU), 5) 17.2, 43 (LPHU + UREA), which is the result of adding 0.4% of the diet DM as urea to LPHU. The corn silage-based treatment diets contained an average of 24% acid detergent fiber and 1.6 Mcal/kg net energy of lactation. Milk urea nitrogen (MUN) concentrations and body weights (BW) were used to calculate predicted amounts of urinary nitrogen (N) using the relationship: urinary N (g/d) = 0.0259 x BW (kg) x MUN (mg/dl). Cows fed HPMU had greater CP and RUP intakes, which resulted in higher concentrations of plasma urea nitrogen, rumen ammonia, MUN, and predicted urinary N. Milk yield, fat yield, fat percent, protein yield, and protein percent were not significantly different among treatments. Parity primarily affected parameters that were related to body size and not measurements of N utilization. The interaction of treatment and parity was not significant for any measurements taken. In this study, cows fed LPHU had significantly lower MUN and predicted urinary N without limiting production. These results demonstrate the potential to optimize milk production while minimizing N excretion in lactating dairy cattle.

Ammonia↗

Measuring the feeding behavior of lactating dairy cows in early to peak lactation.

The objectives of this study were to: 1) objectively define meal criteria (minimum interval between meals) of free-stall housed cows fed via a feed alley, 2) determine which measures of feeding behavior were most repeatable, and 3) describe changes in the feeding behavior from early to peak lactation. An electronic monitoring system was used to record individual cow presence (hits; 6-s resolution) at the feed alley for 21 lactating cows for three 8-d periods: period 1, 35 +/- 16 (mean +/- SD), period 2, 57 +/- 16, and period 3, 94 +/- 16 DIM. A mixture distribution model was used to calculate the meal criterion (27.74 min) by fitting the log10 frequency distribution of the intervals between hits. The within-cow repeatability was highest for feeding activity (hits d(-1)) and intensity (hits per meal min), moderate for total daily mealtime (min d(-1)) and meal duration (min meal(-1)), and lowest for meal frequency (meals d(-1)). From periods 1 to 2, all cows showed increases in total daily mealtime, meal frequency, and meal duration; however, cows with lower meal frequencies and feeding intensity in period 1 showed the greatest increases. Cows with high feeding activity and intensity during period 2 showed proportionally greater increases during period 3. These results illustrate that some measures of feeding behavior are highly repeatable within cows, but variable between cows and across stages of lactation. Thus, tests of treatment effects on feeding behavior should be within cow and control for days in milk.

Animals↗