Effects of concentrate levels on intestinal fermentation and the microbial profile in Japanese draft horses.

In racehorses, feeding a high-concentrate diet could cause abnormal fermentation in the hindgut. This feeding management regime is not suitable for the nutritional physiology of horses. However, studies on the hindgut environment have yet to be reported in Japanese draft horses, so feeding management needs to be investigated in these horses. Therefore, the objective of this study was to investigate the effects of a high-concentrate diet on hindgut fermentation in Japanese draft horses. Feces were collected from 20 male Japanese draft horses managed by two stables with different feeding designs (65% weight ratio of concentrate feed, HC; 50% weight ratio of concentrate, MC), and fecal metabolic characteristics and the microbiome were analyzed. Higher lactate concentrations and lower fecal pH levels were observed in the HC group (P=0.0011, P=0.0192, respectively). Fecal microbiome analysis revealed a decrease in microbial diversity (P=0.0360) and an increase in the relative abundance of Streptococcus lutetiensis/equinus/infantarius (P=0.0011) in the HC group. On the other hand, fibrolytic bacteria in the MC group had similarities with Clostridium sacchalolyticum and Ruminococcus albus. This study revealed that overfeeding of concentrates induced abnormal fermentation in the hindgut of Japanese draft horses. This suggests that the establishment of a feeding design based on not only the chemical compositions of feeds but also microbial dynamics is needed.

Short communication: Serum osteoprotegerin concentrations in periparturient dairy cows.

Dairy cows experience hypocalcemia at the onset of lactation; however, the contribution of bone metabolism to circulating Ca concentrations during the periparturient period is not well understood. Osteoprotegerin (OPG) protects against loss of skeletal mass by regulating osteoclastogenesis. We previously reported that the bone resorption increased around parturition in dairy cows from the perspective of bone markers in blood. The aim of this study was to explore osteoclastic bone resorption in periparturient dairy cows by measuring changes in serum OPG concentrations around calving. The OPG concentrations were evaluated in primiparous (n = 9) and multiparous (n = 9) cows at 21 d precalving and over 5 d after calving. Based on mixed model analyses, the multiparous cows had a higher serum OPG concentration than primiparous cows at precalving and exhibited a significant decline in serum OPG and Ca concentrations after calving. In contrast to multiparous cows, primiparous cows did not exhibit these changes. Therefore, we hypothesized that multiparous cows preserve their bone mineral content during late pregnancy and activate the osteoclastic bone resorption after calving. Based on our serum OPG data, we identified the different periparturient osteoclastic bone resorption patterns in healthy primiparous and multiparous dairy cows.

Relationship of vaginal discharge characteristics evaluated by Metricheck device to metabolic status in postpartum dairy cows.

The relationship between vaginal discharge characteristics and metabolic status has been studied in postpartum dairy cows. Vaginal discharges at 2-6 weeks postpartum were scored weekly on a 1 (clear) to 5 (purulent material with stench) scale using the Metricheck device and classified into three groups as follows: Endometritis group (n = 19), greater than or equal to score 4 until 3 weeks postpartum; Delayed involution group (n = 9), score 2 or 3 after having score 1; Healthy group (n = 23), cows except those belonging to the Delayed involution and Endometritis groups. The uterus and cervix were assessed using ultrasonography. Blood samples were collected twice weekly from 1 to 6 weeks postpartum from all cows and were additionally obtained for 13,14-dihydro-15-keto-prostaglandin F2α (PGFM) measurements from multiparous cows. Cows with delayed involution had delayed onset of luteal activity (p < 0.05) and higher serum urea nitrogen (BUN) concentrations (p < 0.01) than healthy animals. Cows with endometritis had more peripartum diseases (p < 0.05), lower milk yield (p < 0.05), longer cervix diameter (p < 0.05), and lower serum albumin (p < 0.01), total cholesterol (p < 0.01), and magnesium (p < 0.01) concentrations, and higher aspartate aminotransferase levels (p < 0.05) than healthy cows. Plasma PGFM concentrations tended to be higher in the Delayed involution group (p = 0.083) and lower in the Endometritis group (p = 0.085) than in the Healthy group. In summary, delayed involution was shown to be associated with higher BUN concentration, and endometritis may have been caused by peripartum diseases and hepatic dysfunction.

In vivo evaluation of garlic (Allium sativum) supplementation to rice straw-based diet on mitigation of CH4 and CO2 emissions and blood profiles using crossbreed rams.

BACKGROUND: Two experiments were implemented using three cannulated crossbreed rams (Corriedale × Suffolk) fed rice straw-based diet. In vivo evaluation of garlic powder (GP) supplementation to rice straw-based diet on CH4 and CO2 emissions and blood profiles of the rams (body weight = 52 ± 3 kg) was performed. The design was a triplicate 3 × 3 Latin square. Groups were the control (no GP) and two GP groups, which were given supplementation for 0.1 (GP0.1) and 0.5% (GP0.5) of concentrate. The differences between two experiments were data missing of GP0.1 and uncontrolled environment in Experiment 1. Blood profiles were added in Experiment 2. RESULTS: In Experiment 1, supplementation of GP to the rams reduced CH4 production significantly (P < 0.05) by 38%. Dry matter intake was not different among the groups in both experiments. In Experiment 2, CH4 was lower in the GP0.5 group than the control group by 7%, whereas CO2 showed no difference among the groups. Serum glutamic oxaloacetic transaminase was significantly (P > 0.05) lower in the GP0.5 group than the control group, whereas glutamic pyruvic transaminase, total protein, urea N, total cholesterol, low-density lipoprotein, calcium, phosphorus, magnesium and non-esterified fatty acids were not significantly (P > 0.05) different. CONCLUSIONS: GP supplementation, particularly GP0.5, decreases CH4 emissions in rams fed rice straw-based diets. © 2018 Society of Chemical Industry.

Insects as Novel Ruminant Feed and a Potential Mitigation Strategy for Methane Emissions.

This study is the first to evaluate the chemical composition and impacts of four different edible insects, Acheta domesticus (A.d), Brachytrupes portentosus (B.p), Gryllus bimaculatus (G.b), and Bombyx mori (B.m), on the digestibility, rumen fermentation, and methane production when used as a substitute for 25% of the soybean meal (SBM) in a ruminant diet through in vitro incubation. The dietary treatments were 100% grass hay, 60% grass hay + 40% SBM, 60% grass hay + 30% SBM + 10% A.d, 60% grass hay + 30% SBM + 10% B.p, 60% grass hay + 30% SBM + 10% G.b, and 60% grass hay + 30% SBM + 10% B.m. The experiment was conducted as a short-term batch culture for 24 h at 39 °C, and the incubation was repeated in 3 consecutive runs. Chemical analysis of the insects showed that they were rich in fat (14-26%) with a high proportion of unsaturated fatty acids (60-70%). Additionally, the insects were rich in protein (48-61%) containing all essential amino acids and the amino acid profiles of the insects were almost the same as that of SBM. The inclusion of insects did not affect nutrient digestibility or the production of volatile fatty acids but did increase the production of ammonia-nitrogen. The addition of G.b and B.m led to decrease in methane production by up to 18% and 16%, respectively. These results reveal that substitution of 25% SBM in the diet with the tested insects had no negative impacts, and their potential to reduce methane production is an environmental benefit.

The Efficacy of Plant-Based Bioactives Supplementation to Different Proportion of Concentrate Diets on Methane Production and Rumen Fermentation Characteristics In Vitro.

This In Vitro study was conducted to investigate the impact of plant-bioactives extract (PE), a combination of garlic powder and bitter orange extract, on methane production, rumen fermentation, and digestibility in different feeding models. The dietary treatments were 1000 g grass/kg ration + 0 g concentrate/kg ration (100:0), 80:20, 60:40, 40:60, and 20:80. The PE was supplemented at 200 g/kg of the feed. Each group consisted of 6 replicates. The experiment was performed as an In Vitro batch culture for 24 h at 39 °C. This procedure was repeated in three consecutive runs. The results of this experiment showed that supplementation with PE strongly reduced methane production in all kinds of feeding models (p < 0.001). Its efficacy in reducing methane/digestible dry matter was 44% in the 100:0 diet, and this reduction power increased up to a 69.2% with the inclusion of concentrate in the 20:80 diet. The PE application significantly increased gas and carbon dioxide production and the concentration of ammonia-nitrogen, but decreased the pH (p < 0.001). In contrast, it did not interfere with organic matter and fiber digestibility. Supplementation with PE was effective in altering rumen fermentation toward less acetate and more propionate and butyrate (p < 0.001). Additionally, it improved the production of total volatile fatty acids in all feeding models (p < 0.001). In conclusion, the PE combination showed effective methane reduction by improving rumen fermentation characteristics without exhibiting adverse effects on fiber digestibility. Thus, PE could be used with all kinds of feeding models to effectively mitigate methane emissions from ruminants.

Blood markers of osteoclastic differentiation in periparturient dairy cows at different parities, with and without milk fever.

Osteoprotegerin (OPG) inhibits osteoclast (OC) differentiation. TRAP5b (tartrate-resistant acid phosphatase 5b) secreted by OCs reflects the numbers of mature OCs. This study assessed these OC-related markers around parturition in cows of different parities and in cows with milk fever (MF). The blood OPG and TRAP5b concentrations, as well as the ratio of OPG to TRAP5b (O/T), were measured beginning 3 weeks before (-21 d) and over a few days after calving in 49 Holstein Friesian cows at first (n = 8), second (n = 17), third (n = 12), and fourth or greater (n = 12) parities. The ratio of O/T at -21 d to O/T at calving (PreCOT) was also calculated. In the third and greater parities, seven cows developed MF (non-MF, n = 17). Regardless of the development of MF, the serum OPG started to decline in the last week of gestation only in the cows entering the second lactation, while the blood TRAP5b increased at calving in all cows. O/T decreased toward parturition only in multiparous cows. The decrease in O/T at caving was less pronounced in MF cows. PreCOT was negatively correlated with lactation number only in multiparous cows (n = 41, ρ = -0.50, P < .01). This study implied that OC differentiation toward calving was tapered in cows with advanced parities, and these indexes predict the risk of MF.

Effect of parity number on the dry matter intake of dairy cows during the first week after calving.

We investigated the effect of parity number on the dry matter intake (DMI) of cows during the first week after calving. Eighty-three cows were evaluated from 14 days before to 7 days after calving. DMI and milk yield were measured for 7 days after calving, and the calving score was measured. Blood samples were collected throughout the experiment. The average DMI during the first week after calving was reduced in the first-lactation heifers and high-parity number cows. A quadratic relationship between the parity number and the DMI was observed. The first-lactation heifers had lower prepartum serum total protein (TP) concentration and milk yield, higher prepartum serum nonesterified fatty acid (NEFA) concentration and calving score than the multiparous cows. The recovery rate of serum calcium (Ca) after calving was slow in the cows in the parity 6. The DMI was positively affected by the serum Ca concentration after calving, milk yield, and prepartum serum TP concentration and was negatively affected by the calving score and prepartum serum NEFA concentration. We conclude that the DMI immediately after calving tends to be lower in first-lactation heifers and high-parity number cows, but factors that reduce the DMI differ according to parity number.

Impacts of Mootral on Methane Production, Rumen Fermentation, and Microbial Community in an in vitro Study.

Methane mitigation strategies have a two-sided benefit for both environment and efficient livestock production. This preliminary short-term in vitro trial using Mootral (garlic and citrus extracts), a novel natural feed supplement, was conducted to evaluate its efficacy on rumen fermentation characteristics, methane production, and the bacterial and archaeal community. The experiment was performed as a batch culture using rumen fluid collected from sheep, and Mootral was supplemented in three concentrations: 0% (Control), 10%, and 20% of the substrate (50% Grass:50% Concentrate). The rumen fermentation data and alpha diversity of microbial community were analyzed by ordinary one-way analysis of variance. The relative abundance and statistical significance of families and operational taxonomic units (OTUs) among the groups were compared by Kruskal-Wallis H test using Calypso software. After 24-h incubation at 39°C, Mootral in a dose-dependent manner improved the production of total volatile fatty acids and propionate while it reduced the acetate proportion and acetate/propionate ratio. The total produced gas was two times higher in the Mootral-supplemented groups than control (P < 0.01), while the proportion of methane in the produced gas was reduced by 22% (P < 0.05) and 54% (P < 0.01) for 10 and 20% Mootral, respectively. Mootral did not change pH, digestibility, and ammonia-nitrogen. Microbial community analyses showed that Mootral effectively changed the ruminal microbiome. The bacterial community showed an increase of the relative abundance of the propionate-producing family such as Prevotellaceae (P = 0.014) and Veillonellaceae (P = 0.030), while there was a decrease in the relative abundance of some hydrogen-producing bacteria by Mootral supplementation. In the archaeal community, Methanobacteriaceae was decreased by Mootral supplementation compared with control (P = 0.032), while the Methanomassiliicoccaceae family increased in a dose-dependent effect (P = 0.038). The results of the study showed the efficacy of the new mixture to alter the ruminal microbial community, produce more propionate, and reduce microbial groups associated with methane production, thus suggesting that Mootral is a promising natural mixture for methane reduction from ruminants.

Prussian blue in salt blocks decreases radiocesium activity concentration in milk from dairy cattle fed a diet contaminated by the Fukushima nuclear accident.

In Japan, the radiocesium activity concentration in milk must be less than 50 Bq/kg-fresh to meet shipping standards, and the radiocesium concentration of the diet fed to dairy cattle must be less than 500 Bq/kg-dry. After the Fukushima nuclear accident in 2011, we conducted two experiments to investigate whether Prussian blue (PB) could suppress the radiocesium (134Cs + 137Cs) activity concentration in Japanese cattle' milk. In experiment 1, four cattle were fed a diet with a radiocesium activity concentration of 175 Bq/kg-dry, with or without PB supplementation. The PB intake ranged from 0 to 3.0 g/day, and the average radiocesium intake was 3.42 kBq/day in all treatments. The radiocesium activity concentration in milk decreased from 16.4 to 8.6 Bq/kg-fresh, and the transfer coefficient of radiocesium from diet to milk (Fm) decreased from 4.77 × 10-3 to 2.61 × 10-3 with increased PB intake. In experiment 2, three cattle were fed another diet including a radiocesium activity concentration of 927 Bq/kg-dry of with or without PB supplementation. The PB intake ranged from 0 to 18.9 g/day, and the average radiocesium intake was 15.2 kBq/day in all treatments. The milk's radiocesium activity concentration decreased from 24.3 to 4.2 Bq/kg-fresh, and the Fm decreased from 1.68 × 10-3 to 0.28 × 10-3 with increased PB intake. Our results suggest that both the radiocesium activity concentration in milk and Fm can be reduced by PB, and that Fm is affected by diet. We recommend cattle should be fed absorbents such as PB to minimize the risk of milk radiocesium activity concentration exceeding 50 Bq/kg-fresh even if the diet has a radiocesium activity concentration of less than 500 Bq/kg-dry.

Consecutive changes in serum alkaline phosphatase isoenzyme 3 activities in Holstein heifers during the first 18 months of life.

This study investigated consecutive fluctuations in serum activities of bone-specific alkaline phosphatase (ALP) isoenzyme 3 (ALP3) in 11 clinically healthy Holstein heifers during the first 18 months of life. ALP3 activities at the first sampling time point after weaning (3 months) were significantly lower than those at multiple time points during the pre-weaning period. Those activities increased from a minimum at 3 months to a peak at 6 months during the post-weaning period. In the anthropometric data, daily body weight and wither height gains appeared to be below the public data at 4 months and 4-5 months, respectively. The data suggested that serum ALP3 activity can be used to monitor skeletal growth of heifers at weaning.

Influence of calving difficulty on dry matter intake immediately after calving of dairy cows.

We investigated the influence of calving difficulty on dry matter intake (DMI) in Holstein cows immediately after calving using 15 pregnant heifers and 15 multiparous cows. DMI was measured for 6 days after calving. Calving difficulty was evaluated with a calving score, and urinary cortisol was measured. The calving score was higher in the first lactation heifers than in the multiparous cows. The average DMI 6 days after calving was lower in the first lactation heifers than in the multiparous cows. The urinary cortisol concentration at 4 days after calving was higher in the first lactation heifers than in the multiparous cows and was positively associated with the calving score. The DMI was negatively associated with the calving score and the urinary cortisol concentration at 4 days after calving and was positively associated with the average milk yield 6 days after calving and the serum calcium concentration at 3 days after calving. We conclude that DMI immediately after calving is influenced not only by nutrient requirements of cows but also stress derived from calving, and the low DMI in the first lactation heifers is due to the lower energy requirement and high susceptibility to the stress compared with multiparous cows.

The effect of colostrum intake on osteoprotegerin and bone metabolic markers in the blood of newborn calves during the first week of life.

The aim of this study was to elucidate the bone metabolic status after taking colostrum in newborn calves. Fourteen neonatal calves were randomly allocated to two groups fed either unheated or heated (60°C, 30 min) colostrum three times on the first day (2 l every 10 hr; 6 l in total). Heat treatment on colostrum was to reduce the bone metabolic markers assumed as heat-sensitive. The concentrations of four bone metabolic markers (the enzymes from bone cells or the bone collagen fragments) and a bone protective protein, osteoprotegerin (OPG), were measured in the blood of calves during a week after the birth and in the colostrum. The colostral concentrations of four bone metabolic markers were reduced by heating. Then those circulatory markers peaked after colostrum intake in the calves fed unheated colostrum; whereas those fed heated colostrum did not show such changes. However, the plasma tartrate resistant acid phosphatase 5b (TRAP5b) activity was transiently increased after taking colostrum in both groups. Meanwhile, heating did not decrease colostral OPG and there was no significant rise in the serum OPG concentrations after the first colostrum intake in both groups. The study revealed that the blood concentrations of studied bone metabolic markers depended on those colostral values except for TRAP5b. Based on the plasma TRAP5b changes, accelerated formation of premature osteoclast cells may be induced by colostrum intake. Meanwhile, colostral OPG absorption is less likely to impact on its circulating levels.

Effect of substituting soybean meal with euglena (Euglena gracilis) on methane emission and nitrogen efficiency in sheep.

This study evaluated methane (CH4 ) emission, intake, digestibility, and nitrogen efficiency in sheep fed diets containing replacement levels (0%, 33%, 50%, and 67% of soybean meal with euglena). In this experiment, four Corriedale wether sheep with an initial body weight of 53.8 ± 4.6 were arranged in a 4 × 4 Latin square design. This experiment lasted 84 days, divided into four experimental periods. Each period lasted 21 days, which consists of 14 days of adaptation to the diets, 5 days to collect samples, and 2 days to collect gas emission from sheep. Methane emission expressed as L/kg DM intake or g/kg DM intake reduced by up to 37% and the energy loss via CH4 (% of GE intake) reduced by up to 34%. No differences (p > 0.05) were observed in DM and OM intake and whole tract apparent DM digestibility due to substitution of soybean meal with euglena. The total CP loss reduced significantly (linear, p < 0.001) and CP efficiency increased linearly (p = 0.03) with increasing concentration of euglena. As a result, nitrogen balance and average daily weight gain remained unchanged despite higher nitrogen concentration in soybean supplemented group. In conclusion, substitution of soybean meal with euglena reduced methane emission without affecting the performance of animals.