Effect of grass silage harvesting time and level of concentrate supplementation on goat milk quality

Milk fat lipolysis giving high concentrations of free fatty acids (FFA) and off-flavor in the goat's milk is a challenge for the dairy industry in Norway. This has been considered to be caused by underfeeding of the goats and thereby energy mobilization in early and mid lactation. Energy intake can be improved by feeding silage of early harvesting time (HT) and supplementation with concentrate. In the present experiment, 18 goats in early lactation were fed grass silages prepared from the primary growth at a very early, early or normal stage of maturity (HT 1, HT 2 and HT 3, respectively), supplemented with a low (LC; 0.6 kg per goat daily) or normal (NC; 1.2 kg per goat daily) level of concentrate. The experiment was conducted as a cyclic change-over design with four periods of 28 days using three blocks of goats according to their initial body condition (poor, medium or high). Milk and blood samples were collected at the end of each period. Milk yield and yields of milk constituents decreased with delayed harvesting time and with LC. Sensory milk taste quality was not affected by dietary treatment, and milk FFA was highest when NC was fed. The proportion of short and medium chain fatty acids in milk fat decreased with postponed harvesting time and LC, while most of the long chain fatty acids (including C18:1c9) increased with postponed harvesting time and LC. The calculated energy balance decreased and the serum concentration of non-esterified fatty acids (NEFA) increased with decreasing energy content in the diet (postponed harvesting time and low level of concentrate). Goats with initial poor body condition had higher milk FFA concentrations than goats in higher initial body condition. High milk FFA concentration was correlated to poor milk taste quality, low serum NEFA concentration, low C18:1c9 proportion and high energy balance. Our findings suggest that increasing energy intake and energy balance during the first 4 months of lactation does not reduce FFA concentration in goats’ milk.     

Effects of grass silage harvesting time and level of concentrate supplementation on nutrient digestibility and dairy goat performance

The effect of harvesting time (HT) of timothy-dominated grass silage and level of concentrate on the chemical composition of silage, and on feed intake and milk production by Norwegian dairy goats, were evaluated. The silages were prepared from the primary growth at three stages of maturity: very early (HT 1), early (HT 2) and normal (HT 3). The silages were fed ad libitum to 18 goats of the Norwegian dairy goat breed in early lactation and supplemented with a low (LC; 0.6 kg per goat daily) or normal (NC; 1.2 kg per goat daily) level of concentrate. The experiment was conducted as a cyclic changeover design with four periods of 28 days using three blocks of goats according to their initial body condition (poor, medium or high body condition). Silages contained 771, 696 and 619 g digestible organic matter per kg dry matter in silage (D-value) for HT 1, 2 and 3, respectively. Postponing the harvesting time decreased (P<0.001) silage dry matter intake (DMI) and silage DMI per kg body weight (BW). Increased concentrate allowance decreased silage DMI, with substitution rates (decrease in silage DMI when concentrate dry matter intake is increased, kg/kg) of 0.43, 0.21 and 0.27 at HT 1, HT 2 and HT 3, respectively. Milk yield and yields of milk constituents decreased (P<0.001) with delayed harvesting time and thus reflected the changes in silage D-value. Milk free fatty acids (FFA) concentration was not affected by dietary treatments. The efficiency of nutrient utilization was best when LC was fed and increased with postponed harvesting time. The higher energy efficiency of the HT 3 LC fed goats indicates that these goats canalized a higher proportion of energy intake to milk production, compared to goats fed NC and earlier harvested silage. Marginal ECM production response to increased net energy lactation (NEL) intake were higher when intake was increased due to higher silage digestibility (0.14 kg ECM/MJ NEL) compared with increased NEL intake due to increased concentrate level (0.12 kg ECM/MJ NEL). Improving silage quality by earlier harvesting time resulted in higher feed intake and milk yield than obtained by the same increase in NEL intake by concentrate supplementation.     

Effect of roughage type and concentrate supplementation on follicle numbers and in vitro fertilisation and development of oocytes recovered from beef heifers

Increasing dietary energy tends to decrease the ovulatory response and produce fewer viable embryos following superovulation of beef cattle. Data in sheep indicate that high energy intake can decrease progesterone concentrations (P4), although effects in cattle are not as clear. The objectives were to evaluate the effects of roughage type and concentrate supplementation on P4 concentrations, follicle growth and subsequent oocyte fertilisation and embryo development in vitro. Forty-two beef heifers were allocated to 3 treatment groups: (i) silage ad libitum plus 6 kg concentrates (silage + conc.; n = 14); (ii) silage ad libitum (silage; n = 14) or (iii) hay ad libitum (hay; n = 14) for 40 days. Oestrus was synchronised using a controlled intravaginal progesterone releasing device (CIDR) for 7 days plus prostaglandin F2 alpha (15 mg luprostiol) administered 2 days before CIDR withdrawal. Ovaries were stimulated with 600 i.u. of follicle stimulating hormone (pFSH) administered in 6 equal doses at 12-h intervals, starting 12 days after CIDR withdrawal. Daily blood samples were collected from 3 days after CIDR insertion until CIDR withdrawal, and for another 3 days prior to pFSH, for P4 determination. Oocytes were recovered postmortem 12 h after the last pFSH injection, matured, fertilised and cultured in vitro. There was no overall effect of diet (P > 0.05) on P4 concentrations. The number of follicles grown in heifers on silage + conc (18.8 +/- 3.3), silage (23.5 +/- 3.4) or hay (18.1 +/- 2.6) were not affected by the dietary treatment (P > 0.05). The percentage of oocytes fertilised from heifers on hay (88%) was higher compared to oocytes from heifers on silage (79%; P < 0.05), but was not different (P > 0.05) compared to the proportion of oocytes from heifers on silage + conc. (86%). The percentage of fertilised oocytes that cleaved was higher from heifers on silage (94%; P < 0.01) compared with oocytes from heifers on hay (82%) or silage + conc. (86%). The proportion of embryos that developed to blastocyst was not different (P > 0.05) between groups of oocytes from heifers on silage + conc. (8%), silage (14%) or hay (15%). Heifers on silage produced numerically more blastocysts (silage: 19 from 14 heifers; silage + conc.: 8 from 14 heifers; hay: 12 from 14 heifers). These results suggest that dietary treatment used prior to oocyte recovery did not significantly influence the developmental competence of the oocytes in vitro.     

An analysis of the volatile flavour compounds in a soft raw goat milk cheese

The major components of the musty cow rind cheeses were identified in a soft raw goat milk cheese as heptan-2-one, nonan-2-one, their corresponding secondary alcohols, some esters and sulfur compounds. Their production was associated with the manufacturing process and its influence on the microbial activity. However a specificity in goat cheese compounds was displayed concerning in particular limonene and some ketones, alcohols and aldehydes.     

Effect of different levels of concentrate on ruminal microorganisms and rumen fermentation in Nellore steers

The aim of this study was to investigate the effect of different dietary levels of concentrate on feed intake, digestibility, ruminal fermentation and microbial population in steers. Eight Nellore steers fitted with ruminal cannulas were used in a double 4 × 4 Latin square design experiment. The dietary treatments consist of four different proportions of concentrate to roughage: 30:70, 40:60, 60:40 and 80:20% in the dry matter, resulting in Diets 30, 40, 60 and 80, respectively. The roughage was corn silage, and the concentrate was composed of corn, soybean meal and urea. Apparent digestibility of organic matter and crude protein showed a linear association with concentrate proportion (p = 0.01), but the increased concentrate levels did not affect the digestibility of fibre. The lowest ruminal pH-values were observed in animals fed with Diet 80, remaining below pH 6.0 from 6 h after feeding, while in the other diets, the ruminal pH was below 6.0 not before 12 h after feeding. After feeding Diet 80, the ammonia concentration in the rumen was significantly the highest. Higher dietary concentrate levels resulted in a linear increase of propionic acid concentrations, a linear reduction of the ratio acetic acid to propionic acid (p < 0.01) and a linear increased synthesis of microbial nitrogen (p < 0.001). The predicted production of methane was lower in diets with greater amounts of concentrate (p = 0.032). The population of methanogens, R. flavefaciens and R. albus decreased with higher concentrate levels, while the population of S. ruminantium increased (p < 0.05). The results indicate that greater amounts of concentrate do not decrease ruminal pH-values as much as expected and inhibit some cellulolytic bacteria without impairing the dry matter intake and fibre digestibility in Nellore steers.     

Effects of folic acid supplementation to rations differing in the concentrate to roughage ratio on ruminal fermentation, nutrient flow at the duodenum, and on serum and milk variables of dairy cows

The present study was undertaken to determine the effects of dietary folic acid (FOL) supplementation on ruminal fermentation, duodenal nutrient flow, serum and milk variables, and on B-vitamin concentration in serum. The study was divided into two experiments: in Exp. 1 the forage to concentrate (F:C) ratio of the diet (DM basis) was 34:66 (high concentrate, HC), while in Exp. 2 the F:C ratio was 66:34 (high forage, HF). In addition, the cows received 0 or 1 g FOL/d. In Exp. 1, two German Holstein cows equipped with cannulas in the dorsal sac of the rumen and in the proximal duodenum were dry and five were lactating (186 +/- 144 days in milk); in Exp. 2 four cows were dry and four were lactating (165 +/- 57 days in milk). In cows fed the HC diet, FOL supplementation decreased the ruminally-fermented organic matter. Thus, less energy was available for ruminal microorganisms, which resulted in a reduced microbial crude protein flow at the duodenum. Feeding the HF diet, FOL supplementation only increased the apparent ruminal digestibility of acid detergent fibre (ADF). With the HF diet, FOL had no influence on the serum levels of glucose, non-esterified fatty acids, beta-hydroxybutyrate, urea, thiamine, riboflavin, pyridoxal-5'-phosphate, pyridoxic acid, pyridoxal, pyridoxine, pyridoxamine, pantothenic acid, nicotinamide or nicotinic acid, whereas supplementing FOL to the HC diet lowered the serum glucose and riboflavin levels. In both experiments, the supplementation of FOL had no effects on milk composition. Folic acid supplementation to both diets increased the concentrations of serum 5-methyl-tetrahydrofolate. However, no beneficial effects to dairy cows were obvious. Therefore, to achieve certain results, studies with a higher number of non-fistulated cows would be necessary.     

Effects of folic acid supplementation to rations differing in the concentrate to roughage ratio on ruminal fermentation,nutrient flow at the duodenum,and on serum and milk variables of dairy cows

The present study was undertaken to determine the effects of dietary folic acid (FOL) supplementation on ruminal fermentation, duodenal nutrient flow, serum and milk variables, and on B-vitamin concentration in serum. The study was divided into two experiments: in Exp. 1 the forage to concentrate (F:C) ratio of the diet (DM basis) was 34:66 (high concentrate, HC), while in Exp. 2 the F:C ratio was 66:34 (high forage, HF). In addition, the cows received 0 or 1 g FOL/d. In Exp. 1, two German Holstein cows equipped with cannulas in the dorsal sac of the rumen and in the proximal duodenum were dry and five were lactating (186 ± 144 days in milk); in Exp. 2 four cows were dry and four were lactating (165 ± 57 days in milk). In cows fed the HC diet, FOL supplementation decreased the ruminally-fermented organic matter. Thus, less energy was available for ruminal microorganisms, which resulted in a reduced microbial crude protein flow at the duodenum. Feeding the HF diet, FOL supplementation only increased the apparent ruminal digestibility of acid detergent fibre (ADF). With the HF diet, FOL had no influence on the serum levels of glucose, non-esterified fatty acids, beta-hydroxybutyrate, urea, thiamine, riboflavin, pyridoxal-5′-phosphate, pyridoxic acid, pyridoxal, pyridoxine, pyridoxamine, pantothenic acid, nicotinamide or nicotinic acid, whereas supplementing FOL to the HC diet lowered the serum glucose and riboflavin levels. In both experiments, the supplementation of FOL had no effects on milk composition. Folic acid supplementation to both diets increased the concentrations of serum 5-methyl-tetrahydrofolate. However, no beneficial effects to dairy cows were obvious. Therefore, to achieve certain results, studies with a higher number of non-fistulated cows would be necessary.     

Effects of dietary concentrate composition and linseed oil supplementation on the milk fatty acid profile of goats

Milk fat composition can be modulated by the inclusion of lipid supplements in ruminant diets. An interaction between the lipid supplement and the forage to concentrate ratio or the type of forage in the rations may affect milk fat composition. However, little is known about the effects of the starch-to-non-forage NDF ratio in the concentrate and lipid supplementation of goat diets. The aim of this work was to determine the role of dietary carbohydrates in goats rations supplemented with linseed oil on animal performance and milk fatty acid (FA) profile. A total of 16 dairy goats were allocated to two simultaneous experiments (two treatments each), in a crossover design with four animals per treatment and two experimental periods of 25 days. In both experiments alfalfa hay was the sole forage and the forage to concentrate ratio (33:67) remained constant. The concentrate in experiment 1 consisted of barley, maize and soybean meal (concentrate rich in starch), whereas it included soybean hulls replacing 25% of barley and 25% maize in experiment 2 (concentrate rich in NDF). As a result, the starch-to-non-forage NDF ratio was 3.1 in experiment 1 and it decreased to 0.8 in experiment 2. Both concentrates were administered either alone or in combination with 30 g/day of linseed oil. Animal performance parameters were not affected by experimental treatments. In contrast, major changes were observed in milk FA profile due to lipid supplementation and the type of concentrate. Linseed oil significantly raised vaccenic and rumenic acids as well as α-linolenic acid and its biohydrogenation intermediates while decreased medium-chain saturated FA (12:0 to 16:0) in milk fat. Milk fat contents of odd and branched-chain FA and trans-10 18:1 responded differently to linseed oil supplementation according to the concentrate fed.     

Effect of different diluents on goat semen fertility

Skim milk (SM) is considered to be the most widely employed extender for goat sperm used for artificial insemination (AI). However, the fertilizing life span of sperm stored in milk or milk-based extenders does not exceed 12h. Besides some seminal plasma components, such as a protein fraction from the goat bulbourethral gland secretion (SBUIII), interacts with some milk fractions and inhibits the spermatozoa motility. The aim of this study was to prolong the survival of buck semen and its fertility. Buck ejaculates were diluted to a final concentration of 100x10(6)spermatozoa/ml with three different diluents: SM, TEMPOL (4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl) and TEMPOL+hyaluronic acid (TEMPOL+HA). At 7h from dilution 42 goats were inseminated with semen diluted with SM (short-term semen) while after storage for 24h, 44 and 45 goats were inseminated with semen diluted with TEMPOL and TEMPOL+HA (long-term storage), respectively. At day 50 from AI the percentages of pregnant goats were 71.4% (30/42) with SM, 61.4% (27/44) with TEMPOL and 48.8% (22/45) with TEMPOL+HA, with significant differences between SM and TEMPOL+HA. The kidding rate was 66.7% (28/42) with SM diluent, 61.4% (27/44) with TEMPOL and 48.8% (22/45) with TEMPOL+HA, without significant differences among treatment groups. In conclusion, it is possible to maintain good fertility in goats after AI with semen stored for 24h in TEMPOL.     

Effect of different dietary energy level intakes on efficiency of estrus synchronization and fertility in Mashona goat does

The objective of the study was to determine the effects of three dietary energy levels: 0.27 (low level: LL); 0.53 (medium level: ML), and 1.06 (high level: HL) MJ ME kg⁻¹ W^0.75 on estrus synchronization and fertility in Mashona goat does. Forty-five multiparous Mashona goat does of average bodyweight 19.9±2.5 kg were randomly allocated in equal numbers to the three dietary energy levels. The diets were made from a complete feed ration providing 9.83 MJ ME kg⁻¹ DM and 15.5% CP kg⁻¹ DM. Does were fed initially during a 60-day pre-synchronization period, and blood samples were collected twice a week for the determination of plasma progesterone concentrations to ascertain ovarian activity. Intramuscular injections of cloprostenol (100 μg each) were administered 11 days apart. Immediately after the second injection of cloprostenol, three fertile bucks were introduced to the does and were left with the does for 21 days. The does were maintained on their dietary treatments throughout gestation except for those does in the LL treatment. Pregnancy was diagnosed 90 days post-mating using an ultrasound scanner. After pregnancy diagnosis, does on the LL treatment were randomly allocated to ML (n=7) and HL (n=8) treatments. During the pre-synchronization period, does on the LL treatment lost 12.3% whereas those on ML and HL treatments gained 2.1 and 28.8% of their initial bodymasses, respectively. The proportion of does exhibiting overt estrus within 96 h after the last cloprostenol injection was significantly lower (P<0.05) for does on the LL treatment (60%) than for those on ML (93%) or HL (100%) treatments, respectively. However, based on plasma progesterone concentrations, the percentage of does on the LL treatment that exhibited ovarian cycles was numerically lower than that of does that were bred (40 versus 73%). Conception, fecundity and twinning rates were significantly lower (P<0.05) on the LL treatment than on the ML and HL treatments. These results indicate that feeding Mashona goat does 0.27 MJ ME kg⁻¹ W^0.75 compared to 0.53 and 1.06 MJ ME kg⁻¹ W^0.75 reduces the expression of estrus, conception, fecundity and twinning rates, and that feeding 0.53 MJ ME kg⁻¹ W^0.75 suffices for optimum reproduction. In addition, the results suggest that cloprostenol administration may induce ovarian cycles in reproductively quiescent does on dietary energy restriction.     

Effect of vitamin E supplementation on development of reproductive organs in Boer goat

The aim of this study was to evaluate the supplementation of Vitamin E in diet on development of reproductive organs in Boer goat. Twenty-four healthy, Boer male kids of similar body weight (BW) were selected at 3 months of age from the kid flock. The Boer kids were randomly divided into four groups and each group was supplemented with Vitamin E at 0, 80, 320 or 880 IU kid(-1)d(-1) for 5 months. Three Boer goats in each group were slaughtered at the age of 8 months and measured for testis and epididymises and a ductus deferens biopsy by H&E stained slides was run. The results showed that supplementing 80 and 320 IU kid(-1)d(-1) Vitamin E increased weights of the epididymis and the numeric density of convoluted seminiferous tubules. Diameters of convoluted seminiferous tubules and epididymis ductules and numeric density of spermatogenic cells tended to be larger in 80 IU kid(-1)d(-1) compared to the control group (P<0.05). Weight of ductus deferens and thickness of epididymis wall were increased significantly with 320 IU kid(-1)d(-1) (P<0.01). However, no significant effects were observed on thickness of germinal epithelium, numeric density of leydig cells, testis weight or indexes of ductus deferens (P>0.05). In conclusion, supplementing 80 and 320 IU kid(-1)d(-1) Vitamin E in diets significantly simulates the development of reproductive organs in Boer goats. No significantly improved effects were found from supplementing 880 IU kid(-1)d(-1).     

Influence of weather conditions on milk production and rectal temperature of Holsteins fed two levels of concentrate

Twelve lactating Holstein cows in 2nd lactation were allocated randomly, six each, to two feeding treatments: high concentrate (1 kg dairy concentrate to 2 kg milk produced) and low concentrate (1 kg dairy concentrate to 4 kg milk produced) from 7 to 106 days postcalving. Forage and water were provided adalibitum. Milk and butter fat yields and rectal temperatures were examined in relation to 9 weather variables (minimum, maximum and mean temperatures, relative humidity, temperature-humidity index (THI), radiation, wind velocity and mean temperature of the previous day). Averages for milk yield, fat yield and rectal temperature were respectively 20.4 kg, 0.7 kg and 38.9°C for the high concentrate treatment and 18.4 kg, 0.6 kg and 38.6°C for the low concentrate treatment. Weather conditions accounted for 5.6%, 0.8% and 10.8% of the day to day variation in milk yield, fat yield and rectal remperature, respectively, for the high concentrate group and 29.4%, 9.7% and 0.6%, respectively, for the low concentrate group. Only measures of ambient temperature, especially mean temperature, were closely associated with these traits.     

Effect of macromolecule supplementation during in vitro maturation of goat oocytes on developmental potential

In vitro maturation (IVM) of goat oocytes with serum-supplemented media results in oocytes with reduced developmental potential. The objective of this study was to develop a defined medium for IVM of goat oocytes that better supports subsequent embryonic development. Cumulus oocyte complexes (COC) were matured for 18-20 hr in: Experiment (1), tissue culture medium 199 (TCM199) with 10% (v/v) goat serum or modified synthetic oviduct fluid maturation medium (mSOFmat) with 2.5, 8.0, or 20.0 mg/ml bovine serum albumin (BSA); Experiment (2), mSOFmat with 4.0, 8.0, 12.0, or 16.0 mg/ml BSA; or Experiment (3), 1.0 mg/ml polyvinyl alcohol (PVA; control), 4.0 mg/ml BSA, 0.5 mg/ml hyaluronate plus 0.5 mM citrate, or hyaluronate, citrate, and BSA. Mature COC were coincubated for 20-22 hr with 12-15 x 10(6) sperm/ml in modified Brackett and Oliphant (mBO) medium. Embryos were cultured for a total of 7 days in G1/2, and evaluated for cleavage, and blastocyst development, hatching, and total cell numbers. In the first experiment, more (P < 0.05) blastocysts developed per cleaved embryo following maturation in mSOFmat with 2.5 or 8.0 mg/ml BSA than with 20.0 mg/ml BSA or TCM199 with 10% goat serum. The various concentrations of BSA used in the second experiment did not affect (P > 0.05) any of the developmental endpoints examined. In the third experiment, developmental potential of oocytes matured with PVA or hyaluronate with citrate was not different (P > 0.05) from oocytes matured in the presence of BSA. These results demonstrate that developmentally competent goat oocytes can be matured under defined conditions.     

Evidence for existence of lectins on the ruminal bacteria from steers fed roughage and concentrate diets

Mixed rumen bacteria, isolated by centrifugation from the rumen of steers fed a roughage (R) or concentrate (C) diet, were used to determine if lectins are present on rumen bacteria, based on hemagglutination (HA) and HA inhibition assays in vitro. Rumen bacteria from steers fed either diet agglutinated erythrocytes from cattle, sheep, pigs, and rats, suggested that lectins exist on rumen bacteria. Bacterial HA titers from steers receiving the R diet were much higher (p<0.001) than those from steers fed the C diet, depending on the erythrocyte source used. Centrifugation at 20,000xg for 30 min fractionated the rumen bacteria into upper (U) and lower (L) layers. The HA titers of the U bacterial fractions were significantly higher (p<0.001) than those of the L fractions. A remarkable reduction or complete disappearance of HA titers following treatment of rumen bacteria with protease, trypsin, sodium dodecyl sulfate (SDS) or sodium periodate indicates that rumen bacterial lectins are probably glycoproteins. Lectin specificity for saccharides (galactose, lactose, N-acetyl-D-galactosamine, methyl-alpha-D-galactopyranoside and methyl-beta-D-galactopyranoside) and glycoproteins (mucin, fetuin, and thyroglobulin) was found in the RU, RL, and CU bacterial fractions; no specific binding was determined in the CL fractions. The potential role of lectins in mediating the attachment of rumen bacteria to feed particles, rumen epithelia and other microorganisms is discussed.     

Effects of niacin supplementation and dietary concentrate proportion on body temperature,ruminal pH and milk performance of primiparous dairy cows

The objective of this study was to investigate the effects of niacin and dietary concentrate proportion on body temperature, ruminal pH and milk production of dairy cows. In a 2 × 2 factorial design, 20 primiparous Holstein cows (179 ± 12 days in milk) were assigned to four dietary treatments aimed to receive either 0 or 24 g niacin and 30% (low) or 60% (high) concentrate with the rest being a partial mixed ration (PMR) composed of 60% corn and 40% grass silage (on dry matter basis). Ambient temperature and relative humidity were determined and combined by the calculation of temperature humidity index. Respiration rates, rectal, skin and subcutaneous temperatures were measured. Milk production and composition were determined. Ruminal pH and temperature were recorded at a frequency of 5 min using wireless devices for continuous intra-ruminal measurement (boluses). pH values were corrected for pH sensor drift. The climatic conditions varied considerably but temporarily indicated mild heat stress. Niacin did not affect skin, rectal and subcutaneous temperatures but tended to increase respiration rates. High concentrate reduced skin temperatures at rump, thigh and neck by 0.1–0.3°C. Due to the technical disturbances, not all bolus data could be subjected to statistical evaluation. However, both niacin and high concentrate influenced mean ruminal pH. High concentrate increased the time spent with a pH below 5.6 and ruminal temperatures (0.2–0.3°C). Niacin and high concentrate enhanced milk, protein and lactose yield but reduced milk fat and protein content. Milk fat yield was slightly reduced by high concentrate but increased due to niacin supplementation. In conclusion, niacin did not affect body temperature but stimulated milk performance. High concentrate partially influenced body temperatures and had beneficial effects on milk production.     

Effect of different levels of NADH availability on metabolic fluxes of Escherichia coli chemostat cultures in defined medium

Escherichia coli overexpressing a NAD(+)-dependent formate dehydrogenase (FDH) from Candida boidinii was grown in chemostat culture on various carbon sources at 0.05 h(-1) dilution rate, under anaerobic conditions using defined medium and compared to a control without the heterologous FDH pathway. Metabolic fluxes, NADH/NAD(+) ratios and NAD(H/(+)) levels were determined under a range of intracellular NADH availability. The effect of NADH manipulation on the distribution of metabolic fluxes in E. coli was assessed under steady-state conditions. The heterologous FDH pathway converts 1 mol of formate into 1 mol of NADH and carbon dioxide, in contrast with the native FDH where no cofactor involvement is present. Previously, we found that this NADH regeneration system doubled the maximum yield of NADH from 2 to 4 mol NADH/mol glucose consumed and reached 4.6 mol NADH/mol of substrate when sorbitol was used as a carbon source in a complex medium. In the current study, it was found that higher NADH yields and NADH/NAD(+) ratios were achieved with our in vivo NADH regeneration system compared to a control lacking the new FDH pathway in the three carbon sources (glucose, gluconate and sorbitol) examined suggesting a more reduced intracellular environment. The total NAD(H/(+)) amounts were very similar for all the combinations studied. It was also found that the ethanol to acetate ratio increased with increased NADH availability. This ratio increased from 1.05 for the control strain in glucose to 9.45 for the strain expressing the heterologous NAD(+)-dependent FDH in sorbitol.     

不同肥力棕壤全氮和微生物量氮对外源玉米残体氮的响应

以棕壤玉米长期连作定位试验(27a)形成的高低两种肥力水平土壤为研究对象,采用~(15)N标记的玉米植株为试验试材,分别向两种土壤中加入玉米根、茎、叶(共8个处理),采用室内模拟培养与~(15)N同位素示踪技术,旨在弄清玉米根、茎、叶添加后不同肥力土壤全氮含量及微生物量氮的变化规律。结果表明:(1)添加玉米根、茎、叶后低肥力棕壤全氮含量提升幅度分别比高肥力棕壤高5.75%、4.77%和3.75%,外源新氮的贡献率分别比高肥力棕壤高3.54%、3.28%和2.49%,说明不同肥力土壤对玉米残体添加的响应程度不同,低肥力棕壤对外源新氮施入后的响应更敏感,固定能力更强。(2)在添加玉米残体的56d培养时间内,低肥力棕壤中微生物量氮平均增加0.83—0.98倍,高肥力棕壤中微生物量氮平均增加0.87—1.56倍,可以看出不同部位玉米植株添加后均能显著促进土壤微生物量氮的积累,说明外源有机物输入是刺激土壤微生物数量和活性的重要因素,并且在高肥力土壤中刺激作用更加显著。此外,高肥力土壤添加茎和叶处理微生物量氮显著高于根添加处理,但低肥力土壤中根、茎和叶添加处理土壤中微生物量氮之间无显著差异。外源有机氮输入对土壤氮库的贡献与土壤的肥力水平及不同残体部位自身的物质组成特性密切相关。     

Influence of roughage/concentrate ratio and linseed oil on the concentration of trans-fatty acids and conjugated linoleic acid in duodenal chyme and milk fat of late lactating cows

The objective of the study was to investigate the influence of two roughage-to-concentrate ratios, with or without linseed oil supplementation, on the flow of fatty acids in the intestinal chyme and the secretion in milk fat in late lactating cows. Seven late lactating cows fitted with cannulae in the dorsal rumen and simple T-shaped cannulae in the proximal duodenum were randomly assigned to four experimental periods applying an incomplete replicated 2 x 2 Latin square design. The rations consisted of meadow hay and a concentrate mixture given in a ratio of 70:30 or 30:70 on dry matter basis. The basal rations were fed without or with 200 g linseed oil daily. After three weeks of adaptation, samples from the duodenal chyme were taken to study the flow of fatty acids. Additionally, milk samples were analysed for their milk fat composition. Decreasing roughage/concentrate ratio and linseed oil supplementation significantly increased the flow of monounsaturated fatty acids (MUFA), trans-fatty acids (tFA) and conjugated linoleic acids (CLA) in the duodenum. Furthermore, linseed oil increased the flow of saturated fatty acids (SFA) in the duodenum. Higher concentrate portion (H 30) and linseed oil supplementation significantly decreased the milk fat content. SFA were lower (p < 0.05) and MUFA were higher (p < 0.05) in milk fat after linseed oil supplementation; H 30 resulted in more polyunsaturated fatty acids (PUFA, p < 0.05) in the milk. Linseed oil supplementation significantly increased tFA and CLA in milk fat. The higher CLA content in milk fat as compared to that in the digesta suggests that a substantial endogenous synthesis of CLA in the mammary gland tissue through A9-desaturase took place. Between 21% and 48% of duodenal t11-C(18:1) were converted into c9, t11-CLA in milk fat.