Untargeted metabolomic investigate milk and ruminal fluid of Holstein cows supplemented with Perilla frutescens leaf.

Milk compounds are important for human nutrient requirements and health. The ruminal metabolic profile is responsible for dietary nutrition and determines milk production. Perilla frutescens leaf (PFL) is a commonly used medicinal herb due to its bioactive metabolites. This study elucidated the effects of PFL on the metabolome of two biofluids (rumen fluid and milk) of 14 cows fed a basic total mixed ration diet (CON, n = 7) and supplemented with 300 g/d PFL per cow (PFL, n = 7) by ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry. Milk PE-NMe (18:1(9Z)/18:1(9Z)) and DG (18:0/20:4(5Z,8Z,11Z,14Z)/0:0), oleanolic acid, and nucleotides were upregulated, and milk medium-chain fatty acids (2-hydroxycaprylic acid) were down-regulated in response to PFL. The supplementation of PFL increased the abundance of pyrimidine nucleotides both in rumen fluid and milk. The pathways of pyrimidine metabolism and biosynthesis of unsaturated fatty acids were enriched both in the rumen fluid and milk. We also found the milk 2-hydroxycaprylic acid was positively correlated with ruminal uridine 5-monophosphate, and was negatively correlated with ruminal deoxycytidine, and the milk thymidine was positively correlated with ruminal icosenoic acid. This study found that the supplementation of PFL could alter the ruminal metabolic profiles and milk synthesis through regulation of the pathways of pyrimidine metabolism and biosynthesis of unsaturated fatty acids. Our new findings provide comprehensive insights into the metabolomics profile of rumen fluid and milk, supporting the potential production of Perilla frutescens milk in dairy cows.

Effect of Broussonetia papyrifera L. silage on blood biochemical parameters, growth performance, meat amino acids and fatty acids compositions in beef cattle.

OBJECTIVE: The study was conducted to investigate the effects of Broussonetia papyrifera L.(B. papyrifera) silage on growth performance, serum biochemical parameters, meat quality, and meat amino acids and fatty acids compositions in beef cattle. METHODS: Sixty-four male Angus beef cattle were assigned to 4 groups with 4 pens in each group and 4 beef cattle in each pen, and fed with the total mixed ration supplemented with 0%, 5%, 10%, or 15% B. papyrifera silage for 100 days (control group, 5% group, 10% group and 15% group) separately. RESULTS: Beef cattle had significantly higher final body weight (BW) in 15% group, higher average daily gain (ADG) and dry matter intake (DMI) in 5% group, 10% group and 15% group, and higher feed conversion ratio (FCR) in 10% group and 15% group. Significantly higher blood superoxide dismutase (SOD) concentration was noted in 15% group, higher blood total antioxidant capacity (TAC) in 10% group and 15% group, lower 8-hydroxydeoxyguanosine (8-OHdG) and malondialdehyde (MDA) in 15% group. Meat had lower pH in 15% group, higher Commission International DeI'Eclairage (CIE) L* in 5% group, 10% group, and 15% group, and lower drip loss in 15% group. Greater concentration of meat polyunsaturated fatty acids (PUFA) was observed in 10% group and 15% group, and docosahexaenoic acid (DHA) in 15% group. CONCLUSION: Diet with 15% B. papyrifera silage could improve performance and increase final BW, ADG, DMI, and FCR, enhance the antioxidant functions by decreasing blood 8-OHdG and MDA and increasing blood SOD and TAC, improve the meat quality by lowing pH and drip loss and increasing CIE L*, increase the meat PUFA and DHA concentration. Polyphenols and flavonoids might be the main components responsible for the antioxidant activity and anti-biohydrogenation in the B. papyrifera silage. And B. papyrifera silage could be used as a new feedstuff in beef cattle nutrition.

Rumen Fermentation Characteristics in Pre- and Post-Weaning Calves upon Feeding with Mulberry Leaf Flavonoids and Candida tropicalis Individually or in Combination as a Supplement.

Although flavonoids or yeast have been used as feed additives to improve the production efficiency and health of adult cattle, little information is available on their effects on rumen fermentation in calves. The objective of this study was to investigate the effects of feed supplementation with mulberry leaf flavonoids and Candida tropicalis on performance, blood parameters, and rumen fermentation characteristics during pre-weaning and post-weaning periods. Forty-eight Holstein calves were used in a four-groups trial and were supplemented with (1) no yeast or flavonoids (CON), (2) active dry yeast (ADY; C. tropicalis, 5.0 × 109 CFU/d), (3) flavonoids (FLA; 3 g/d), or (4) yeast and flavonoids (YF; C. tropicalis, 5.0 × 109 CFU/d; flavonoids, 3 g/d). The feeding strategy was as follows: milk replacer was offered at 12% of body weight in two meals per calf each day at age 21 d, and a gradual weaning process was adopted at age 50 to 56 days. Data of daily feed intake, body weight, and serum and rumen fermentation parameters were obtained at 28, 42, 56, and 80 d ages, respectively. A significant time effect and interaction between treatment and time was found for average daily gain, feed efficiency, total volatile fatty acid concentration, and proportion of propionate in calves (p < 0.05). Average daily gain and feed efficiency increased during post-weaning and overall periods for calves in FLA and YF groups compared with CON and ADY groups (p < 0.05). A reduction of fecal scores with supplementation was found in FLA and YF groups (p < 0.05). Rumen fluid pH and ammonia nitrogen concentration remained constant across the groups, whereas total volatile fatty acid concentration and molar proportion of propionate significantly increased during the pre-weaning and overall periods in FLA and YF groups (p < 0.05). Calves in YF group had the highest serum concentrations of IgG and IgA during the overall period (p < 0.05). Additionally, serum ß-hydroxybutyric acid concentration was higher in ADY and FLA groups during the post-weaning period (p < 0.05). Supplementation with C. tropicalis showed little effect on increasing growth performance and health compared with flavonoids alone. Meanwhile, the combination of C. tropicalis and flavonoids was not synergistic with respect to improving health and rumen fermentation compared with use of flavonoids alone in pre- and post-weaning calves (p > 0.05).

Effects of Tea Saponin Supplementation on Nutrient Digestibility, Methanogenesis, and Ruminal Microbial Flora in Dorper Crossbred Ewe.

Two experiments were conducted using Dorper × thin-tailed Han crossbred ewes. In experiment 1, eighteen ewes were randomly assigned to two dietary treatments (a basal diet, or the same basal diet supplemented with 2.0 g tea saponin (TS)/head/day) to investigate the effects of TS supplementation on nutrient digestibility and methane emissions. In experiment 2, six ewes with ruminal cannulae were assigned to the same two dietary treatments as in experiment 1 to investigate the effects of TS supplementation on rumen fermentation and microbial flora. TS supplementation increased the apparent digestibility of organic matter (OM) (p = 0.001), nitrogen (N) (p = 0.036), neutral detergent fibre (NDF) (p = 0.001), and acid detergent fibre (ADF) (p < 0.001). Urinary N (p = 0.001) and fecal N (p = 0.036) output were reduced, and N retention (p = 0.001) and nitrogen retention/nitrogen intake (p = 0.001) were increased. Supplementary TS did not decrease absolute methane emissions (p = 0.519) but decreased methane emissions scaled to metabolic bodyweight by 8.80% (p = 0.006). Ammonia levels decreased (p < 0.001) and total volatile fatty acid levels increased (p = 0.018) in response to TS supplementation. The molar proportion of propionate increased (p = 0.007), whereas the acetate:propionate ratio decreased (p = 0.035). Supplementation with TS increased the population of Fibrobacter succinogenes (p = 0.019), but the population of protozoans tended to decrease (p = 0.054). Supplementation with TS effectively enhanced the apparent digestibility of OM, N, NDF, and ADF, and decreased methane emissions scaled to metabolic bodyweight.

Effect of Broussonetia papyrifera L. (paper mulberry) silage on dry matter intake, milk composition, antioxidant capacity and milk fatty acid profile in dairy cows.

OBJECTIVE: This study was carried out to investigate the possible application of Broussonetia papyrifera (B. papyrifera) silage as a functional feeding stuff in dairy cattle. METHODS: Seventy-two Holstein cows were divided into four groups randomly and allocated to 6 pens with 3 individuals in each group and fed the original total mixed ratio (TMR) in the dairy farm or the new TMR with 5%, 10%, and 15% B. papyrifera silage, separately. Feed intake were recorded, milk and blood samples were collected, and milk composition, blood metabolites and milk fatty acids composition were measure at the end of the experiment. RESULTS: Dry matter intake of cows decreased when they fed on diet with B. papyrifera, but no differences were observed in body condition score, milk yield, milk protein and lactose, feed efficiency and serum metabolites between groups. Both 10% or 15% of B. papyrifera silage in the diet significantly increased the immunoglobulin A (IgA) and IgG in serum, 15% of B. papyrifera silage increased the content of serum catalase, superoxide dismutase, total antioxidant capacity, and decreased the content of 8-hydroxy-2'-deoxyguanosine. Furthermore, 10% or 15% of B. papyrifera silage resulted in a significant decrease in the milk somatic cell count, and increased the polyunsaturated fatty acids content in the milk. CONCLUSION: The diets with 10% to 15% of B. papyrifera silage might enhance the immune and antioxidant function of dairy cows and increase the polyunstaturated fatty acid concentration in the milk.

Effects of feeding a Moringa oleifera rachis and twig preparation to dairy cows on their milk production and fatty acid composition, and plasma antioxidants.

BACKGROUND: We determined how supplementing the diet of lactating, multiparous Holstein dairy cows with a preparation of Moringa oleifera rachises and twigs affected their milk production and quality and the levels of plasma antioxidants. RESULTS: We found that milk yield increased in cows receiving the 6% (w/w) moringa supplement compared with that of the control. Addition of the moringa supplement increased the concentration of milk fat and decreased the somatic cell count in the milk. However, protein, glucose and total solid and urea nitrogen concentrations in the milk were the same for all treatments. The concentration of glutathione peroxidase increased for cows fed the moringa supplement compared with the control. The percentages of total unsaturated fatty acids, mono-unsaturated fatty acids, and polyunsaturated fatty acids including n-3 polyunsaturated fatty acid increased in the milk of cows fed the moringa supplement compared with those of the controls. CONCLUSION: Addition of the moringa supplement into the diet of lactating multiparous cows improved milk production and health status and modified milk fatty acid profile positively. The results suggested that moringa supplement could be used as a diet supplement for producing high quality and healthier milk. © 2017 Society of Chemical Industry.

Effect of weaning age on growth performance, feed efficiency, nutrient digestibility and blood-biochemical parameters in Droughtmaster crossbred beef calves.

OBJECTIVE: The objective of this study was to determine the effect of weaning age on intake, performance, nutrition metabolism and serum parameters of beef calves. METHODS: Sixty Droughtmaster crossbred calves were assigned to 5 groups with 12 calves in each group. The calves in control group remained with the dams till the 22-week age, while the calves weaned at 28 d (4 wk), 42 d (6 wk), 56 d (8 wk), and 70 d (10 wk) of age were sent to group 4 wk, group 6 wk, group 8 wk, and group 10 wk, respectively, and then were fed on milk replacer till the 22-week age. Feed intake and body weight and size were record and blood metabolites were measured. And 24 calves of them (6 in each group) were picked randomly for digestion and metabolism trail. Feed, feces and urine sample were taken and measured. RESULTS: Dry matter intake of calves in group 4 wk was significantly lower than those in the remaining groups from wk 17 to 22 (p<0.05). Feed efficiency of the calves was higher in groups 4 wk and 6 wk than those in groups 8 wk and 10 wk from 11 to 13 wk (p<0.05), and calves had higher feed efficiency in group 4 wk, group 6 wk, and group 8 wk than those in group 10 wk from wk 14 to wk 22. Calves in group 4 wk and 6 wk had lower body weight than group 8 wk and group 10 wk and control group at 10-week age (p<0.05) and 13-week age (p<0.05), and calves in group 6 wk had no significant difference in body weight with control group, group 8 wk and 10 wk (p>0.05) but was higher than that of group 4 wk (p<0.05). Calves in group 6 wk had higher final body weight and total gain than group 4 wk, but no difference of total gain with that of groups 8 wk, 10 wk, and control group. And weaning calves at 6-week age brought higher feed efficiency and average daily gain from wk 14 to wk 22, and higher dry matter and organic matter digestibility at 21 wk. CONCLUSION: It is concluded that the weaning of calves at 6 weeks of age gave positive results.

Net protein and metabolizable protein requirements for maintenance and growth of early-weaned Dorper crossbred male lambs.

BACKGROUND: Dorper is an important breed for meat purpose and widely used in the livestock industry of the world. However, the protein requirement of Dorper crossbred has not been investigated. The current paper reports the net protein (NP) and metabolizable protein (MP) requirements of Dorper crossbred ram lambs from 20 to 35 kg BW. METHODS: Thirty-five Dorper × thin-tailed Han crossbred lambs weaned at approximately 50 d of age (20.3 ± 2.15 kg of BW) were used. Seven lambs of 25 kg BW were slaughtered as the baseline animals at the start of the trial. An intermediate group of seven randomly selected lambs fed ad libitum was slaughtered at 28.6 kg BW. The remaining 21 lambs were randomly divided into three levels of dry matter intake: ad libitum or 70% or 40% of ad libitum intake. Those lambs were slaughtered when the lambs fed ad libitum reached 35 kg BW. Total body N and N retention were measured. RESULTS: The daily NP and MP requirements for maintenance were 1.89 and 4.52 g/kg metabolic shrunk BW (SBW0.75). The partial efficiency of MP utilization for maintenance was 0.42. The NP requirement for growth ranged from 12.1 to 43.5 g/d, for the lambs gaining 100 to 350 g/d, and the partial efficiency of MP utilization for growth was 0.86. CONCLUSIONS: The NP and MP requirements for the maintenance and growth of Dorper crossbred male lambs were lower than the recommendations of American and British nutritional systems.

Dietary supplementation with mulberry leaf flavonoids inhibits methanogenesis in sheep.

The effects of flavonoids on methanogenesis and microbial flora in Dorper × thin-tailed Han crossbred ewes were evaluated in two experiments. To investigate the effects of flavonoids on nutrient digestibility and nitrogen balance, 18 ewes (60.0 ± 1.73 kg body weight (BW)) were allotted to two dietary treatments in experiment one, a control diet and the control diet supplemented with flavonoids (2 g/head/day). In experiment two, the effects of supplementary flavonoids on ruminal fermentation and microbial flora were investigated using quantitative polymerase chain reaction with six ewes (67.2 ± 0.79 kg BW) with ruminal cannula assigned to the identical dietary treatments used in experiment one. Supplementary flavonoids improved the apparent digestibility of nitrogen (N, P < 0.001) and neutral detergent fibre (NDF, P = 0.024) and decreased daily CH4 output (P < 0.001). The ruminal pH (P = 0.638) and ammonia (P = 0.690) were not affected by supplementary flavonoids, whereas the total volatile fatty acid (VFA) content increased (P = 0.037). Supplementary flavonoids decreased ruminal populations of protozoans (P = 0.002) and methanogens (P < 0.001) and increased the populations of Fibrobacter succinogenes (P = 0.016). In conclusion, flavonoids improved the digestibility of organic matter and reduced CH4 output by inhibiting the populations of microbes involved in methanogenesis.

Effects of different source additives and wilt conditions on the pH value, aerobic stability, and carbohydrate and protein fractions of alfalfa silage.

To improve the silage quality and reduce the silage additive cost, the present experiment was designed to evaluate the potential of applying the fermented juice of epiphytic lactic acid bacteria (FJLB) as an additive in alfalfa silage. The effects of FJLB on the fermentation quality, carbohydrate and protein fractions, and aerobic stability of alfalfa silage wilted under five different conditions were investigated and compared with commercial lactic acid bacteria (CLAB) and the control. The FJLB application decreased the pH value, the volatile fatty acids and non-protein nitrogen content, and the loss of sugar by 9.9%, 22.9%, 19.6% and 9.6%, respectively; it increased the lactic acid concentration by 29.5% and the aerobic stability by 17 h in comparison to the control. The FJLB application also decreased the pH value (4.44 vs. 4.66) and volatile fatty acid content (38.32 vs. 44.82) and increased the lactic acid concentration (68.99 vs. 63.29) in comparison to the CLAB-treated silage. However, the FJLB treatment had lower aerobic stability (254 h vs. 274 h) than the CLAB treatment. The FJLB application improved silage quality in comparison to the control; in addition, its effect as a fermentation stimulant may be comparable to or even better than CLAB.

Effect of supplementation of allicin on methanogenesis and ruminal microbial flora in Dorper crossbred ewes.

BACKGROUND: Garlic extracts have been reported to be effective in reducing methanogenesis. Related mechanisms are not well illustrated, however, and most studies have been conducted in vitro. This study investigates the effects of supplementary allicin (AL) in sheep diet on in vivo digestibility, rumen fermentation, and shifts of microbial flora. METHODS: Two experiments were conducted using Dorper × thin-tailed Han crossbred ewes. In experiment 1, eighteen ewes (60.0 ± 1.73 kg BW) were randomly assigned for 29 days to either of two dietary treatments: a basal diet or the basal diet supplemented with 2.0 g AL/head·day to investigate supplementary AL on nutrient digestibility and methane emissions. In experiment 2, six ewes (65.2 ± 2.0 kg BW) with ruminal canulas were assigned to the same two dietary treatments as in experiment 1 for 42 days to investigate supplementary AL on ruminal fermentation and microbial flora. The methane emissions were determined using an open-circuit respirometry system and microbial assessment was done by qPCR of 16S rRNA genes. RESULTS: Supplementary AL increased the apparent digestibility of organic matter (P < 0.001), nitrogen (P = 0.006), neutral detergent fiber (P < 0.001), and acid detergent fiber (P = 0.002). Fecal nitrogen output was reduced (P = 0.001) but urinary nitrogen output was unaffected (P = 0.691), while nitrogen retention (P = 0.077) and nitrogen retention/nitrogen intake (P = 0.077) tended to increase. Supplementary AL decreased methane emissions scaled to metabolic bodyweight by 5.95 % (P = 0.007) and to digestible organic matter intake by 8.36 % (P = 0.009). Ruminal pH was unaffected (P = 0.601) while ammonia decreased (P = 0.024) and total volatile fatty acids increased (P = 0.024) in response to supplementary AL. Supplementary AL decreased the population of methanogens (P = 0.001) and tended to decrease that of protozoans (P = 0.097), but increased the populations of F. succinogenes (P < 0.001), R. flavefaciens (P = 0.001), and B. fibrisolvens (P = 0.001). CONCLUSIONS: Supplementation of AL at 2.0 g/head·day effectively enhanced OM, N, NDF, and ADF digestibility and reduced daily methane emissions (L/kg BW(0.75)) in ewes, probably by decreasing the population of ruminal protozoans and methanogens.

Effects of protein sources for milk replacers on growth performance and serum biochemical indexes of suckling calves.

This study investigated the effects of protein sources for milk replacers on growth performance and serum biochemical indexes of suckling calves. Fifty Chinese Holstein bull calves with similar BW and age were randomly allocated to 5 groups (1 control and 4 treatments) of 10 calves in each group. Five types of milk replacers were designed to have the same level of energy and protein. The protein source for milk replacers of the control group was full milk protein (MP). The protein source of milk replacers of the 4 treatment groups was composed of MP and one vegetable protein (VP) (30 and 70% of total protein). The 4 types of VP were soybean protein concentrate (SP), hydrolyzed wheat protein (WP), peanut protein concentrate (PP), and rice protein isolate (RP). Results of the experiment showed: 1) there was no significant difference on average daily gain (ADG) and feed:gain ratio (F:G) among the MP, SP and RP groups (P > 0.05), whereas the ADG and F:G of the WP and PP groups were significantly lower compared with the MP group (P < 0.05); 2) there was not a significant difference in withers height, body length and heart girth among treatment groups compared with the MP group (P > 0.05). Thereby the 4 VP milk replacers had no adverse effects on body size of calves; 3) all groups showed no significant difference in the serum contents of urea nitrogen, total protein, albumin, globulin, ß-hydroxybutyrate, growth hormone, insulin-like growth factor-1, and the ratio of albumin to globulin (A:G) (P > 0.05). In conclusion, SP or RP (accounts for 70% of the total protein) as calf milk replacers could substitute MP, whereas wheat gluten and PP had a significant adverse effect on growth performance in this experiment.

Effect of Dietary Concentrate:forage Ratios and Undegraded Dietary Protein on Nitrogen Balance and Urinary Excretion of Purine Derivatives in Dorper×thin-tailed Han Crossbred Lambs.

THIS STUDY AIMED TO INVESTIGATE DIETARY CONCENTRATE: forage ratios (C:F) and undegraded dietary protein (UDP) on nitrogen balance and urinary excretion of purine derivatives (PD) in lambs. Four Dorper×thin-tailed Han crossbred castrated lambs with 62.3±1.9 kg body weight at 10 months of age were randomly assigned to four dietary treatments in a 2×2 factorial arrangement of two levels of C:F (40:60 and 60:40) and two levels of UDP (35% and 50% of CP), according to a complete 4×4 Latin-square design. Each experimental period lasted for 19 d. After a 7-d adaptation period, lambs were moved into individual metabolism crates for 12 d including 7 d of adaption and 5 d of metabolism trial. During the metabolism trial, total urine was collected for 24 h and spot urine samples were also collected at different times. Urinary PD was measured using a colorimetric method and creatinine was measured using an automated analyzer. Intake of dry matter (DM) (p<0.01) and organic matter (OM) (p<0.01) increased as the level of UDP decreased. Fecal N was not affected by dietary treatment (p>0.05) while urinary N increased as the level of UDP decreased (p<0.05), but decreased as dietary C:F increased (p<0.05). Nitrogen retention increased as dietary C:F increased (p<0.05). As dietary C:F increased, urinary excretion of PD increased (p<0.05), but was not affected by dietary UDP (p>0.05) or interaction between dietary treatments (p>0.05). Daily excretion of creatinine was not affected by dietary treatments (p<0.05), with an average value of 0.334±0.005 mmol/kg BW(0.75). A linear correlation was found between total PD excretion and PDC index (R(2) = 0.93). Concentrations of creatinine and PDC index in spot urine were unaffected by sampling time (p>0.05) and a good correlation was found between the PDC index (average value of three times) of spot urine and daily excretion of PD (R(2) = 0.88). These results suggest that for animals fed ad libitum, the PDC index in spot urine is effective to predict daily excretion of PD. In order to improve the accuracy of the spot sampling technique, an appropriate lag phase between the time of feeding and sampling should be determined so that the sampling time can coincide with the peak concentration of PD in the urine.