Effects of sunflower oil infusions of Asparagopsis taxiformis on in vitro ruminal methane production and biohydrogenation of polyunsaturated fatty acids.

Asparagopsis taxiformis inhibits ruminal methane (CH4) production due to its bromoform (CHBr3) content. The immersion of A. taxiformis in edible vegetable oils allows the extraction and stabilization of the highly volatile CHBr3 in the oil phase. The objectives of this study were to explore the effects of adding sunflower oils with increasing concentrations of CHBr3 on in vitro ruminal methanogenesis and biohydrogenation. Five batches of 48-h in vitro incubations were performed in 14 fermentation bottles, using rumen inocula collected shortly after the slaughter of young crossbred bulls and 1 g of dry matter (DM) from a total diet of mixed feed without added oil (control) or with 60 µL of sunflower oil per gram of DM as the substrate. The treatments were the CHBr3 content in the oil added: 0 µg (B0), 25 µg (B25), 50 µg (B50), 75 µg (B75), 100 µg (B100), and 150 µg (B150) of CHBr3 per gram of substrate DM. Organic matter (OM) degradability, total gas, CH4, volatile fatty acids (VFA), long-chain fatty acids, and dimethyl acetals (DMA) were analyzed at the end of each incubation. Data were analyzed with a model considering the treatments as the fixed effect and the run as a random block and using orthogonal contrasts. Degradability of OM was higher in the control group and was unaffected by CHBr3 concentration. Total gas production per gram of degraded OM was unaffected by treatments and averaged 205 ± 29.8 mL/g. Methane (mL) production decreased linearly with increasing CHBr3 concentrations, with 33%, 47%, and 87% reductions for B75, B100, and B150, respectively. Total VFA concentration was unaffected by oil inclusion but was reduced by 20% in CHBr3-containing treatments, although without any dose-response pattern. The molar percentage of acetate decreased linearly, whereas propionate and butyrate increased linearly with the increasing CHBr3 dosage. Including oil in the diet decreased the branched-chain fatty acids and DMA content. Increasing CHBr3 concentrations did not affect branched-chain fatty acids, but linearly increased most of the identified DMA. Adding oil to the control diet increased the 18:2n-6, whereas increasing the concentration of CHBr3 had no effect on 18:2n-6 but decreased linearly the 18:0 and increased the trans-18:1 isomers. The results obtained provide evidence that oil immersions of A. taxiformis can successfully inhibit ruminal production of CH4 in vitro at doses of 100 and 150 µg/g DM, and simultaneously modulate biohydrogenation.

Rumen microbial community and milk quality in Holstein lactating cows fed olive oil pomace as part in a sustainable feeding strategy.

The use of alternative feed ingredients from the Agro-industry could be an efficient tool to improve the sustainability of dairy cow production. Since the richness in polyphenols, olive oil pomace (OOP), produced during olive oil milling, seems a promising by-product to ameliorate milk's nutritional value. The aim of this study was to test the use of OOP produced by means of a new technology (biphasic with stone deprivation) in dairy cow feeding strategy to evaluate the effect on animal performances, rumen microbiota, biohydrogenation processes and milk quality by a multidisciplinary approach. Forty multiparous Italian-Friesian dairy cows, at middle lactation, were randomly allotted into two homogenous groups and fed respectively a commercial diet (CON) and the experimental diet (OOPD) obtained by adding OOP to CON as partial replacement of maize silage. The two diets were formulated to be isoproteic and isoenergetic. The same diets were tested also in an in vitro trial aimed to evaluate their rumen degradability (% DEG). The dietary supplementation with OOP did not affect DM intake, rumen % DEG and milk production. The milk's nutritional quality was improved by increasing several important functional fatty acids (FAs; i.e., linoleic acid, conjugated linoleic acid, oleic acid, vaccenic acid). This finding was related to a decrease in rumen liquor biohydrogenation rate of unsaturated FAs. The stochiometric relation between volatile FA production in the rumen and methanogenesis suggested that OOP lowers the methane potential production (CON = 0.050 mol/L vs OOPD = 0.024 mol/L, SEM = 0.005, P = 0.0011). Rumen microbiota and fungi community did not be strongly altered by OOP dietary inclusion because few bacteria were affected at the genus level only. Particularly, Acetobacter, Prevotellaceae_UCG-004, Prevotellaceae_UCG-001, Eubacterium coprostanoligenes, Lachnospira, Acetitomaulatum, Lachnospiraceae_NK3A20 group were more abundant with OOPD condition (P < 0.05). Data reported in this study confirm that the use of OOP in dairy cow feeding can be an interesting strategy to improve milk nutritional quality increasing functional FA content without compromising the rumen degradability of the diet or causing strong perturbation of rumen ecosystem and maintaining animal performances.

Producing natural functional and low-carbon milk by regulating the diet of the cattle-The fatty acid associated rumen fermentation, biohydrogenation, and microorganism response.

Conjugated linoleic acid (CLA) has drawn significant attention in the last two decades for its various potent beneficial effects on human health, such as anticarcinogenic and antidiabetic properties. CLA could be generally found in ruminant products, such as milk. The amount of CLA in ruminant products mainly depends on the diet of the animals. In general, the fat content in the ruminant diet is low, and dietary fat supplementation can be provided to improve rumen activity and the fatty acid (FA) profile of meat and milk. Especially, dietary 18-carbon polyunsaturated FA (C18 PUFA), the dominant fat source for ruminants, can modify the milk FA profile and other components by regulating the ruminal microbial ecosystem. In particular, it can improve the CLA in milk, intensify the competition for metabolic hydrogen for propionate producing pathways and decrease methane formation in the rumen. Therefore, lipid supplementation appears to be a promising strategy to naturally increase the additional nutritional value of milk and contribute to lower methane emissions. Meanwhile, it is equally important to reveal the effects of dietary fat supplementation on rumen fermentation, biohydrogenation (BH) process, feed digestion, and microorganisms. Moreover, several bacterial species and strains have been considered to be affected by C18 PUFA or being involved in the process of lipolysis, BH, CLA, or methane emissions. However, no review so far has thoroughly summarized the effects of C18 PUFA supplementation on milk CLA concentration and methane emission from dairy cows and meanwhile taken into consideration the processes such as the microorganisms, digestibility, rumen fermentation, and BH of dairy cattle. Therefore, this review aims to provide an overview of existing knowledge of how dietary fat affects rumen microbiota and several metabolic processes, such as fermentation and BH, and therefore contributes to functional and low-carbon milk production.

Fresh Phyllanthus emblica (Amla) Fruit Supplementation Enhances Milk Fatty Acid Profiles and the Antioxidant Capacities of Milk and Blood in Dairy Cows.

The objective of this study was to investigate the effect of a diet supplemented with fresh amla fruit as a natural feed additive on blood metabolic parameters, milk antioxidant capacity, and milk fatty acid (FA) proportions in lactating dairy cows. Eight ruminally cannulated mid-lactation dairy cows were used in a repeated crossover design. The first group of four cows received total mixed ration (TMR) feed without fresh amla fruit (control group). The remaining four cows sequentially supplemented fresh amla fruit (FAF) at three levels (200, 400, then 600 g/d) (treatment group) at 14-day intervals. In second period, control and treatment groups were exchanged. The first ten days were adjusted to diet adaptation for each sub-period, and the last four days for sampling milk and blood. A total of 514 metabolites were detected from FAF using UPLC-ESI-MS/MS. The five main metabolites in FAF were phenolic acids (22%), flavonoids (20%), lipids (20%), amino acids and derivatives (9%), and tannins (7%). Amla fruit supplementation reduced total saturated fatty acid and the omega-6/omega-3 ratio at 200 or 400 g/d FAF dose compared to controls. In addition, amla fruit increased unsaturated FA, such as C20:5 (Eicosapentaenoic acid, EPA) and C22:6 (docosahexaenoic acid, DHA), and branched-chain FA in a dose-dependent manner at 200 or 400 g/d compared to controls. In addition, amla fruit increased the antioxidant capacity biomarkers in the blood, such as superoxide dismutase (SOD) and albumin; this confirms that amla fruit is an excellent antioxidant, inhibiting reactive oxygen species' (ROS) metabolism, and can thereby protect cells from oxidative stress. Moreover, the most remarkable improvement of ferric reducing-antioxidant power (FRAP) and total antioxidant capacity (TAC) in milk was recorded at 400 g/d FAF doses compared to controls. Therefore, fresh amla fruit doses for lactating cows at 400 g/d on an as-fed basis can be used as an alternative additive feed in dairy cow diets to improve antioxidant capacity, protein efficiency, butter quality, and to produce more desirable milk fatty acid profiles for human consumption.

Grape seed tannin extract and polyunsaturated fatty acids affect in vitro ruminal fermentation and methane production.

Condensed tannins (CT), one of the most ubiquitous compounds in the plant kingdom, can modulate ruminal nutrient metabolism. Objectives were to study potential interactions of CT and polyunsaturated fatty acids (PUFA) on ruminal fermentation, biohydrogenation (BH), and methane production. Ruminal fluid obtained from lactating Holstein Friesian cows was used. All experiments were carried out as a completely randomized design with the same mixed diet: control (60:40 forage:concentrate) without supplement (CON), 2.5% soybean oil (SBO), and SBO + grape seed tannin extract (GSTE) at 0.2%, 0.4%, 0.6%, or 0.8% dietary DM (ST0.2, ST0.4, ST0.6, and ST0.8, respectively). Compared with CON (84.7 mM), total VFA concentration was not affected by SBO, but decreased (P < 0.001) with ST0.8 vs. ST0.6 (75.3 vs. 78.3 mM). Relative to CON, methane production was depressed (P < 0.001) by 17.7% and 28.0% in ST0.4 and ST0.8. The highest (P < 0.001) mean concentrations of c9,t11 CLA and C18:1 t11 were observed with ST0.4 compared with CON, but there was no difference between SBO and CT-containing diets. Disappearance of C18:2 c9,c12 was 49.1% vs. 50.3% in CON vs. SBO, whereas it ranged from 39.9% to 46.3% in CT-containing diets after 2 h incubation (P < 0.001). Concentrations of c9,t11 CLA with supplemental SBO and ST0.8 nearly peaked (P < 0.001) at 2 h incubation, but this fatty acid peaked (P > 0.05) at 6 h incubation and remained higher (P < 0.001; 15.9-17.0 µg/mL) at 24 h incubation with ST0.2, ST0.4, and ST0.6 compared with other diets (13.5-14.5 µg/mL). Compared with CON (50.6 µg/mL), concentration of C18:1 t11 with SBO and CT-containing diets reached a peak (P < 0.001; 241-265 µg/mL) at 12 h incubation. Concentration of C18:0 was 171%-231% higher (P < 0.001) with SBO and CT relative to CON at 24 h incubation. Overall, these results demonstrated that PUFA in SBO are more effective in modulating ruminal BH and CH4 production when combined with CT. However, high doses of added CT can reduce ruminal VFA concentration. Thus, a level of 0.4% GSTE added to diets containing 2.5% PUFA from plant origin might be suitable for optimizing ruminal fermentation and BH of C18:2 c9,c12 to fatty acid intermediates that could have beneficial effects to human health.

Condensed tannins can modulate methane emissions and ruminal biohydrogenation, but effects depend on type and dose. We used an in vitro fermentation system to investigate the effect of increasing doses (0%, 0.2%, 0.4%, 0.6%, and 0.8% dry matter) of grape tannin seed extract (GSTE) in a diet supplemented at 2.5% dry matter with soybean oil on methane production and biohydrogenation. Feeding soybean oil and GSTE at 0.6% and 0.8% reduced content of ruminal volatile fatty acids. Methane production (mL/g dry matter) was lower in the diet containing GSTE at 0.4%. Inclusion of GSTE at 0.2% and 0.4% increased concentration of C18:2 c9,c112, C18:3n3, c9,t11 conjugated linoleic acid and total polyunsaturated fatty acids after 24 h of incubation. The present findings contribute to a better understanding of the effect of condensed tannins from grape seed extract on ruminal fermentation and biohydrogenation.

Partial substitution of fish oil for linseed oil enhances beneficial fatty acids from rumen biohydrogenation but reduces ruminal fermentation and digestibility in growing goats.

This study was performed to investigate effects of partial replacement of fish oil (FO) for linseed oil (LO) on digestibility, ruminal fermentation and biohydrogenation in growing goats. Experiment 1 was carried out in four growing male goats aged 6 months in a 4 × 4 Latin square design. Goats were fed a basal diet supplemented with 25 g/kg dry matter either LO alone or in combination with tuna FO. Treatments were developed by replacing FO for LO at ratios of 0, 5, 10 and 15 g/kg DM corresponding to FO-0, FO-5, FO-10 and FO-15, respectively. Experiment 2 was carried out in an in vitro incubation system including 12 fermenters with the same four treatments. Each fermenter consisted of 40 mL goat ruminal fluid, 160 mL warm buffer, 2 g mixed substrates, and 50 mg FO-0, FO-5, FO-10 or FO-15. Fish oil inclusion reduced (P < 0.05) digestibility and nitrogen retention in Experiment 1. Increasing doses of FO in the diet induced a strong drop (P < 0.001) in ruminal total volatile fatty acid (VFA) concentration and protozoa population at 3 h post incubation, but did not affect individual VFA proportions. Substitution of FO for LO decreased mean concentrations of C18:0 (P = 0.057), c-9,c-12 C18:2 and C18:3n-3 (P < 0.001), but increased (P < 0.001) C20:5n-3 and C22:6n-3. Feeding FO-10 enhanced formation of ruminal c-9,t-11 conjugated linoleic acid (CLA) concentration compared with FO-0. Overall, combined data suggest that to improve ruminal concentrations of C20:5n-3, C22:6n-3, and c-9,t-11 CLA for deposition in tissues or milk with minimal risk of affecting digestibility and ruminal fermentation, a dietary supplementation of 15 g/kg LO and 10 g/kg FO would be suitable.

Inclusion of cocoa by-product in the diet of dairy sheep: Effect on the fatty acid profile of ruminal content and on the composition of milk and cheese.

In this study, we hypothesized that dietary cocoa bean shell (CBS) as a partial replacer of human edible cereal grains in the diet of lactating ewes may affect performance and milk and cheese composition. Twenty Comisana lactating ewes allotted into control (CTRL; n = 10) or cocoa (CBS; n = 10) group received alfalfa hay ad libitum and 800 g of conventional (CTRL) or experimental (CBS) concentrate containing 11.7% CBS to partially replace corn and barley of the CTRL concentrate. Milk yield and composition did not differ between groups, and only urea concentration was lower in CBS milk. Dietary CBS increased cheese fat and reduced protein percentage in CBS group. Fatty acid composition of rumen content partially reflected that of the ingested diet, with total saturated fatty acids (SFA), total monounsaturated fatty acids (MUFA), 16:0, 18:0 and 18:1c9 greater in the CBS group. Moreover, all the identified trans- and cis-18:1 isomers were greater in CBS rumen content. Milk and cheese showed a similar fatty acid composition. Total MUFAs were greater in milk and cheese of CBS, mainly due to the proportion of 18:1c9, and conversely, total polyunsaturated fatty acids (PUFA), PUFAn-6 and PUFAn-6-to-PUFAn-3 ratio was greater in CTRL group. Concluding, the inclusion of CBS in the diet of lactating ewes within the limit imposed by the current legislation did not cause detrimental effects on animal performance and milk composition. Interestingly, dietary CBS reduced milk urea concentration probably due to the phenols contained in CBS concentrate. However, our results support that biohydrogenation was weakly impaired by dietary CBS. Finally, CBS negatively affected cheese nutritional characteristics due to lower protein and greater fat content, but improved fat health indexes in milk and cheese.

Influence of yeast on rumen fermentation, growth performance and quality of products in ruminants: A review.

This review aims to give an overview of the efficacy of yeast supplementation on growth performance, rumen pH, rumen microbiota, and their relationship to meat and milk quality in ruminants. The practice of feeding high grain diets to ruminants in an effort to increase growth rate and weight gain usually results in excess deposition of saturated fatty acids in animal products and increased incidence of rumen acidosis. The supplementation of yeast at the right dose and viability level could counteract the acidotic effects of these high grain diets in the rumen and positively modify the fatty acid composition of animal products. Yeast exerts its actions by competing with lactate-producing (Streptococcus bovis and Lactobacillus) bacteria for available sugar and encouraging the growth of lactate-utilising bacteria (Megasphaera elsdenii). M. elsdenii is known to convert lactate into butyrate and propionate leading to a decrease in the accumulation of lactate thereby resulting in higher rumen pH. Interestingly, this creates a conducive environment for the proliferation of vaccenic acid-producing bacteria (Butyrivibrio fibrisolvens) and ciliate protozoa, both of which have been reported to increase the ruminal concentration of trans-11 and cis-9, trans-11-conjugated linoleic acid (CLA) at a pH range between 5.6 and 6.3. The addition of yeast into the diet of ruminants has also been reported to positively modify rumen biohydrogenation pathway to synthesise more of the beneficial biohydrogenation intermediates (trans -11 and cis -9, trans -11). This implies that more dietary sources of linoleic acid, linolenic acid, and oleic acid along with beneficial biohydrogenation intermediates (cis-9, trans-11-CLA, and trans-11) would escape complete biohydrogenation in the rumen to be absorbed into milk and meat. However, further studies are required to substantiate our claim. Therefore, techniques like transcriptomics should be employed to identify the mRNA transcript expression levels of genes like stearoyl-CoA desaturase, fatty acid synthase, and elongase of very long chain fatty acids 6 in the muscle. Different strains of yeast need to be tested at different doses and viability levels on the fatty acid profile of animal products as well as its vaccenic acid and rumenic acid composition.

Combined effects of 3-nitrooxypropanol and canola oil supplementation on methane emissions, rumen fermentation and biohydrogenation, and total tract digestibility in beef cattle.

The individual and combined effects of 3-nitrooxypropanol (3-NOP) and canola oil (OIL) supplementation on enteric methane (CH4) and hydrogen (H2) emissions, rumen fermentation and biohydrogenation, and total tract nutrient digestibility were investigated in beef cattle. Eight beef heifers (mean body weight ± SD, 732 ± 43 kg) with ruminal fistulas were used in a replicated 4 × 4 Latin square with a 2 (with and without 3-NOP) × 2 (with and without OIL) arrangement of treatments and 28-d periods (13 d adaption and 15 d measurements). The four treatments were: control (no 3-NOP, no OIL), 3-NOP (200 mg/kg dry matter [DM]), OIL (50 g/kg DM), and 3-NOP (200 mg/kg DM) plus OIL (50 g/kg DM). Animals were fed restrictively (7.6 kg DM/d) a basal diet of 900 g/kg DM barley silage and 100 g/kg DM supplement. 3-NOP and OIL decreased (P < 0.01) CH4 yield (g/kg DM intake) by 31.6% and 27.4%, respectively, with no 3-NOP × OIL interaction (P = 0.85). Feeding 3-NOP plus OIL decreased CH4 yield by 51% compared with control. There was a 3-NOP × OIL interaction (P = 0.02) for H2 yield (g/kg DM intake); the increase in H2 yield (P < 0.01) due to 3-NOP was less when it was combined with OIL. There were 3-NOP × OIL interactions for molar percentages of acetate and propionate (P < 0.01); individually, 3-NOP and OIL decreased acetate and increased propionate percentages with no further effect when supplemented together. 3-NOP slightly increased crude protein (P = 0.02) and starch (P = 0.01) digestibilities, while OIL decreased the digestibilities of DM (P < 0.01) and neutral detergent fiber (P < 0.01) with no interactions (P = 0.15 and 0.10, respectively). 3-NOP and OIL increased (P = 0.04 and P < 0.01, respectively) saturated fatty acid concentration in rumen fluid, with no interaction effect. Interactions for ruminal trans-monounsaturated fatty acids (t-MUFA) concentration and percentage were observed (P = 0.02 and P < 0.01); 3-NOP had no effect on t-MUFA concentration and percentage, while OIL increased the concentration (P < 0.01) and percentage (P < 0.01) of t-MUFA but to a lesser extent when combined with 3-NOP. In conclusion, the CH4-mitigating effects of 3-NOP and OIL were independent and incremental. Supplementing ruminant diets with a combination of 3-NOP and OIL may help mitigate CH4 emissions, but the decrease in total tract digestibility due to OIL may decrease animal performance and needs further investigation.

Meta-analysis of the relationship between milk trans-10 C18:1, milk fatty acids <16 C, and milk fat production.

The economic value of milk fat and its responsiveness to management strategies provides strong interest in maximizing milk fat production by minimizing occurrence of biohydrogenation-induced milk fat depression (BH-MFD) and maximizing de novo synthesized fatty acids (FA). Tools that allow a timely diagnosis of BH-MFD would improve nutritional management. Specific milk FA or FA categories correlate to milk fat concentration and are of interest for diagnosing the cause of changes in milk fat concentration. The objective of the current study was to characterize the relationship between milk fat concentration and trans-10 C18:1, a proxy for BH-MFD, and FA <16 carbons that originate solely from de novo lipogenesis using a meta-analysis approach that used data from the literature and unpublished Penn State experiments. Prior to the meta-analysis, the effect of FA methylation method on milk FA profile was tested to determine potential bias between papers. There was no difference between sodium methoxide, acid, and acid-base methylation methods on trans-10 C18:1 concentration, but acid methods resulted in loss of short-chain FA. The relationship between trans-10 C18:1 and milk fat percentage was investigated using a 2-component model, where one component described the fraction unresponsive to BH-MFD and the other described a responsive fraction that is exponentially related to trans-10 C18:1. The 2 fractions where characterized utilizing a Bayesian hierarchical model accounting for between-study variability. The model was defined by the function f(x, θ1, θ2, θ3) = θ1 + θ2exp(θ3), where the unresponsive θ1 fraction was 2.15 ± 0.09%, the responsive θ2 fraction was 1.55 ± 0.08%, and the exponential term θ3 was -0.503 ± 0.07 (posterior mean ± posterior standard deviation from the Bayesian hierarchical model). A Lin's concordance correlation coefficient of 0.67 suggested good agreement between observations and predictions from the Bayesian hierarchical model, computed only with the model's mean population parameters. There was a linear relationship between milk fat concentration and FA <16 C as a percentage of total FA (intercept = 2.68 ± 0.237 and slope = 0.043 ± 0.011; coefficient of determination = 0.31). The relationship between milk FA <16 C and milk fat concentration is weaker than what has been published, likely because multiple factors can reduce de novo FA without reducing milk fat and the broad range of diets present in the literature.

Invited review: Role of rumen biohydrogenation intermediates and rumen microbes in diet-induced milk fat depression: An update.

To meet the energy requirements of high-yielding dairy cows, grains and fats have increasingly been incorporated in ruminant diets. Moreover, lipid supplements have been included in ruminant diets under experimental or practical conditions to increase the concentrations of bioactive n-3 fatty acids and conjugated linoleic acids in milk and meat. Nevertheless, those feeding practices have dramatically increased the incidence of milk fat depression in dairy cattle. Although induction of milk fat depression may be a management tool, most often, diet-induced milk fat depression is unintended and associated with a direct economic loss. In this review, we give an update on the role of fatty acids, particularly originating from rumen biohydrogenation, as well as of rumen microbes in diet-induced milk fat depression. Although this syndrome seems to be multi-etiological, the best-known causal factor remains the shift in rumen biohydrogenation pathway from the formation of mainly trans-11 intermediates toward greater accumulation of trans-10 intermediates, referred to as the trans-11 to trans-10 shift. The microbial etiology of this trans-11 to trans-10 shift is not well understood yet and it seems that unraveling the microbial mechanisms of diet-induced milk fat depression is challenging. Potential strategies to avoid diet-induced milk fat depression are supplementation with rumen stabilizers, selection toward more tolerant animals, tailored management of cows at risk, selection toward more efficient fiber-digesting cows, or feeding less concentrates and grains.

Effect of 2-hydroxy-4-(methylthio) butanoate (HMTBa) supplementation on rumen bacterial populations in dairy cows when exposed to diets with risk for milk fat depression.

Diet-induced milk fat depression (MFD) is a condition marked by a reduction in milk fat yield experimentally achieved by increasing dietary unsaturated fatty acids and fermentable carbohydrates. 2-Hydroxy-4-(methylthio) butanoate (HMTBa) is a methionine analog observed to reduce diet-induced MFD in dairy cows. We hypothesize that the reduction in diet-induced MFD by HMTBa is due to changes in the rumen microbiota. To test this, 22 high-producing cannulated Holstein dairy cows were placed into 2 groups using a randomized block design and assigned to either control or HMTBa supplementation (0.1% of diet dry matter). All cows were then exposed to 3 different diets with a low risk (32% neutral detergent fiber, no added oil; fed d 1 to 7), a moderate risk (29% neutral detergent fiber and 0.75% soybean oil; fed d 8 to 24), or a high risk (29% neutral detergent fiber and 1.5% soybean oil; fed d 25 to 28) for diet-induced MFD. Rumen samples were collected on d 0, 14, 24, and 28, extracted for DNA, PCR-amplified for the V1-V2 region of the 16S rRNA gene, sequenced on an Illumina MiSeq (Illumina, San Diego, CA), and subjected to bacterial diversity analysis using the QIIME pipeline. The α diversity estimates (species richness and Shannon diversity) were decreased in the control group compared with the HMTBa group. Bacterial community composition also differed between control and HMTBa groups based on both weighted UniFrac (relative abundance of commonly detected bacteria) and unweighted UniFrac (presence/absence) distances. Within the HMTBa group, no differences were observed in bacterial community composition between d 0 and d 14, 24, and 28; however, in the control group, d 0 samples were different from d 14, 24, and 28. Certain bacterial genera including Dialister, Megasphaera, Lachnospira, and Sharpea were increased in the control group compared with the HMTBa group. Interestingly, these genera were positively correlated with milk fat trans-10,cis-12 conjugated linoleic acid and trans-10 C18:1, fatty acid isomers associated with biohydrogenation-induced MFD. It can be concluded that diet-induced MFD is accompanied by significant alterations in the rumen bacterial community and that HMTBa supplementation reduces these microbial perturbations.

Combination of pelleting and monensin does not affect antioxidant properties and fatty acids in milk of grazing dairy cows supplemented with a concentrate containing soybean seeds.

This study was performed with the main objective of evaluating the effect of the combination of pelleting and monensin on fatty acids (FA) composition, the concentration of total polyphenols and flavonoids, and the oxidative stability of milk in cows fed a concentrate containing soybean seeds. Eight Holstein multiparous cows were distributed in a replicated Latin square design. The four supplement treatments consisted of the combination of two factors (pelleting and monensin) and one concentrate as follows: (1) unpelleted concentrate with no monensin (CO); (2) pelleted concentrate with no monensin (PE); (3) unpelleted concentrate with 96 mg of monensin/kg of dry matter, DM (MO); and (4) pelleted concentrate with 96 mg of monensin/kg of DM (PM). There was no interaction between pelleting and monensin for milk production and concentration of milk protein, lactose, total polyphenols, flavonoids, conjugated dienes (CD), and reducing power. Fat and total solids concentration in milk were decreased when cows were fed pelleted (PE and PM) concentrates. Feeding cows with PE and PM concentrates increased the CD concentration in milk. Regarding milk FA concentration, there was no difference among treatments for total saturated, monounsaturated, and polyunsaturated FA. The most prominent result was that pelleting increased the milk concentration of omega-3 FA. Altogether, the present study suggests that the pelleting process can improve the milk fat quality by increasing the omega-3 FA, while the combination of pelleting and monensin in the diet of grazing dairy cows fed soybean-based concentrate adds no further improvements to FA profiles and oxidative stability of milk.

Effect of stearic or oleic acid on milk performance and energy partitioning when fed in diets with low and high rumen-active unsaturated fatty acids in early lactation.

This experiment was conducted to determine the effects of stearic acid (SA; C18:0) or rumen-protected oleic acid (OA; C18:1 cis-9) on milk performance and energy partitioning of early lactation cows when supplemented in diets with low and high level of rumen unsaturated fatty acids (RUFA). In low RUFA experiment (LRUFA), FA supplement rich in either SA or calcium salts OA was added to a basal diet with a low concentration of RUFA (0.75% vs. 1.4%, LRUFA-SA vs. LRUFA-OA). In high RUFA experiment (HRUFA), 2% soybean oil was added to the diet fed in the LRUFA experiment. In each experiment, 30 multiparous cows were blocked by parity and predicted transmitting ability for milk yield and were randomly fed 1 of 2 treatment diets from 2 to 13 wk postpartum. In the LRUFA experiment, LRUFA-SA had 2.4 kg/d more dry matter intake (DMI) (P < 0.01), 3.8 kg/d more energy-corrected milk (P < 0.01), and 0.3% units more milk fat percentage (P < 0.01) and 0.2 kg/d more milk fat yield (P < 0.01). Dietary treatments did not affect body weight, energy balance, and energy intake partitioning into milk, maintenance, and body tissues (P > 0.1). In the HRUFA experiment, HRUFA-SA had 1.4 kg/d more DMI (P = 0.03) but similar milk and milk components yields (P > 0.1). HRUFA-SA had a tendency to gain more body weight (P = 0.07) and had more positive energy balance (P = 0.01) and decreased gross feed efficiency (milk yield/DMI) (P = 0.01). Consistently, HRUFA-SA increased intake energy partitioning into body tissues (P = 0.02) and decreased energy partitioning into milk (P = 0.01). In summary, SA supplementation had more DMI relative to OA, but the effects on milk and milk fat production were different and affected by the level of RUFA in the basal diet. In application, SA supplementation was more effective to improve milk production when included in the basal diet with the low RUFA.

Culture pH interacts with corn oil concentration to affect biohydrogenation of unsaturated fatty acids and disappearance of neutral detergent fiber in batch culture.

Effects of culture pH and corn oil (CO) concentration on biohydrogenation (BH) of unsaturated fatty acids and disappearance of neutral detergent fiber (NDF) in batch culture were evaluated in a 2 × 3 factorial design experiment. Culture vessels (100 mL; 4 replicates/treatment per time point) included ground alfalfa hay plus CO at 0, 1, or 2% dry matter inclusion rate and were incubated at pH 5.8 (low pH) or 6.2 (high pH) for 0, 6, 12, 18, or 24 h. Effects of culture pH, CO, time, and their interactions were determined. Adding CO increased total fatty acid concentration in substrates to 1.01, 2.31, and 3.58% dry matter for 0, 1, and 2% CO, respectively. Corn oil concentration interacted with culture pH and resulted in different effects on BH of cis-9,cis-12 18:2 at low or high culture pH. After 24 h of incubation, low pH, compared with high pH, reduced disappearance of NDF by 35% and BH extent of cis-9,cis-12 18:2 by 31%. Increasing CO increased disappearance of NDF across pH treatments and decreased BH extent of cis-9,cis-12 18:2 at low pH and increased it at high pH over 24 h. Compared with high pH, low pH reduced concentrations of 18:0 by 31% and increased concentrations of trans-10,cis-12 18:2 and trans-10 18:1 by 110 and 79% after 24 h, respectively. Adding CO at low pH had greater effect on BH intermediates of cis-9,cis-12 18:2 compared with adding oil at high pH. In particular, increasing CO to 1 and 2% DM at low pH, compared with at high pH, resulted in a 36 and 46% reduction in the concentration of 18:0, an 84 and 131% increase in the concentration of trans-10,cis-12 18:2, and an 81 and 129% increase in the concentration of trans-10 18:1, respectively. Despite the interactions between culture pH and CO concentration, main effects across time were also significant for the response variables of interest. In conclusion, culture pH interacted with CO concentration to affect BH of UFA and disappearance of NDF in batch culture, as the effects were greater at low culture pH than at high culture pH.

The effects of dietary quercetin supplementation and sex on the fatty acid profile of rabbit meat, dissectible fat and caecotrophes.

This study examined the effects of dietary quercetin on the fatty acid (FA) profile of rabbit caecotrophes, dissectible fat, Longissimus thoracis et lumborum (LTL) muscle and hindleg (HL) meat. Sixteen male and sixteen female New Zealand White rabbits were fed a control or quercetin-supplemented (2 g quercetin dihydrate/kg feed) diet from 5 to 12 weeks old, then slaughtered. Caecotrophes were collected from the gut, and the dissectible fat, LTL and deboned HL were sampled. Lipids in the samples were transmethylated, then identified and quantified using GC-FID. Quercetin-supplementation increased C18:0 in the fat, and C20:4n-6 in the LTL - suggesting an interaction with endogenous lipid metabolism - but had no effect on the HL and caecotrophes. Sex affected the caecotrophe FAs, but had little effect on the meat's nutritional value. The FA profiles of the LTL and HL differed, but both aligned to nutritional recommendations. The caecotrophe FA profile was indicative of microbial biohydrogenation, but this had minimal effect on the carcass FA.

Milk fat response and milk fat and urine biomarkers of microbial nitrogen flow during supplementation with 2-hydroxy-4-(methylthio)butanoate.

2-Hydroxy-4-(methylthio)butanoate (HMTBa) is a methionine analog that has been observed to attenuate biohydrogenation (BH)-induced milk fat depression (MFD), possibly through reducing the shift to altered BH pathways. It has also been suggested that HMTBa increases microbial protein synthesis in the rumen. Our objectives were to stimulate BH-induced MFD and (1) verify HMTBa inhibition of BH-induced MFD and changes in milk fatty acids (FA) associated with altered rumen BH (i.e., trans-10 C18:1); and (2) determine the effect of HMTBa on milk fat (i.e., odd- and branched-chain FA) and urine biomarkers related to microbial N flow. Twenty-four multiparous cows (45.6 ± 8.5 kg of milk/d; mean ± standard deviation) and 12 primiparous cows (32.8 ± 3.1 kg of milk/d) were arranged in a crossover design. Treatments were unsupplemented control and HMTBa fed at 0.1% of diet dry matter intake. The experiment was 80 d and included a 10-d pretrial covariate period. Each experimental period included 2 phases that differed in risk for BH-induced MFD, including a 28-d low-risk phase (31.6% neutral detergent fiber, 21.8% starch, and no oil) and a 7-d moderate-risk phase (28.7% neutral detergent fiber, 28.1% starch, and 1.0% soybean oil). We found no interaction of treatment and parity. Milk fat yield (1.43 ± 0.51 kg/d) and milk fat trans-10 C18:1 (0.42 ± 0.08 g/100 g of FA) did not differ between treatments during the low-risk phase. However, during the moderate-risk phase, HMTBa maintained higher milk fat concentration (3.91 vs. 3.79%), tended to maintain higher milk fat yield (1.44 vs. 1.38 kg/d), and decreased milk fat trans-10 C18:1 (0.61 vs. 0.93% FA) compared with control. Additionally, HMTBa increased milk fat concentration and secretion of odd- and branched-chain FA by 5.3 and 10.2%, respectively, but urinary biomarkers of microbial N flow (i.e., purine derivatives) did not differ between treatments. However, rumen bacterial samples were not available to provide cow- or treatment-specific microbial protein-to-marker ratios, which is a critical source of variation. Additionally, transfer of odd- and branched-chain FA to milk is dependent on several factors that may affect interpretation of these biomarkers. In conclusion, HMTBa decreased absorption of alternate BH intermediates and maintained higher milk fat when feeding a diet with moderate-risk for MFD.

Changes in fermentation and animal performance during recovery from classical diet-induced milk fat depression using corn with differing rates of starch degradability.

Diet-induced milk fat depression (MFD) is a multifactorial disorder that can be triggered by a variety of conditions. Feeding high amounts of starch and unsaturated fatty acids has been shown to reduce milk fat yield and composition, as well as alter ruminal biohydrogenation patterns. However, little is known about how starch degradability in the rumen influences recovery from diet-induced MFD and if production of milk fat-inhibiting isomers will persist following an episode of MFD. The objective of this study was to evaluate production performance and ruminal fermentation in cows recovering from MFD when corn with a low or high starch degradability is fed. Six ruminally fistulated Holstein cows were used in a crossover design with 2 periods. During each period, MFD was induced for 10 d by feeding a diet with low fiber, high starch, and high unsaturated fatty acid. The polyunsaturated fatty acid concentration of the diet during the induction phase was modified primarily through inclusion of soybean oil. Following induction, cows were switched to either a high degradable starch recovery diet (HDS) or a low degradable starch recovery diet (LDS) for 18 d. The 7-h starch degradability was 66.5% for LDS and 87.8% for HDS. Milk was collected every 3 d for component and fatty acid analysis. On d 0, 4, 7, 10, 16, 22, and 28 of each period, ruminal pH and rumen fluid were collected every 2 h. Milk fat yield and composition was reduced during MFD induction and progressively increased by day in both HDS and LDS during recovery. Dry matter intake was similar among treatments and increased steadily over time during recovery. Preformed fatty acids were greater for HDS-fed animals, and de novo fatty acid in milk fat was greater for LDS-fed animals. Milk trans-10 C18:1 tended to be greater for HDS, and trans-10,cis-12 conjugated linoleic acid was significantly greater for HDS. cis-9,trans-11 conjugated linoleic acid was not affected by starch degradability during recovery. Total volatile fatty acids, butyrate, and valerate tended to differ or differed with recovery treatment, but ruminal pH and ammonia concentration were unaffected. The HDS diet responded similarly to the LDS diet during recovery with regard to milk fat percentage, but milk and fat yield tended to consistently be lower in HDS. When considering approaches to ameliorate diet-induced MFD, the degradability of the starch within rations should be evaluated. Although animal performance was similar, some trans fatty acid isomers were persistent in the milk through the recovery phase with HDS-fed animals, suggesting that milk fat synthesis might be potentially inhibited and biohydrogenation pathways modified in the rumen following an episode of MFD.

Linseed oil and heated linseed grain supplements have different effects on rumen bacterial community structures and fatty acid profiles in cashmere kids1.

This study investigated the effects of dietary supplementation with alternative sources of α-linolenic acid on growth, the composition of rumen microbiota, and the interactions between rumen microbiota and long-chain fatty acid (FA) concentrations, in goat kids. Sixty 4-month-old castrated male Albas white cashmere kids (average BW 18.6 ± 0.1 kg) were randomly allocated among three dietary treatments: (i) basal diet without supplementation (Control), (ii) basal diet supplemented with linseed oil (LSO), (iii) basal diet supplemented with heated linseed grain (HLS). The concentrate:forage ratio was 5:5 and the LSO and HLS treatments provided the kids with similar dietary FA profiles. The diets were fed for 104 d, consisting of 14 d for adaptation followed by 90 d of experimental observation. Treatment did not significantly influence BW, DMI, or bacterial richness or diversity. On the other hand, the relative abundance of bacteria participating in hydrogenation differed significantly among the three groups: the Veillonellaceae and Christensenellaceae were more abundant in LSO kids, Prevotellaceae were more abundant in HLS kids, and the Fibrobacteriaceae were more abundant in Control kids (P < 0.05). Spearman correlation analysis indicated that Ruminobacter, Selenomonas_1, Fretibacterium, Prevotellaceae_UCG-001, Succinimonas, and Ruminococcaceae_NK4A214_group were the genera that participated in hydrogenation of long-chain FAs. HLS-fed kids had a lower relative abundance of Ruminobacter, but a higher abundance of Prevotellaceae_UCG-001 and Fretibacterium than LSO-fed kids. These changes were associated with greater rumen concentrations of C18:3n3 and n-3 PUFA, but lower concentrations of n-6 PUFA and lower n-6/n-3 ratios, in HLS than in LSO-fed kids. In conclusion, feeding kids with HLS increased rumen concentrations of C18:3n3 and n-3 PUFA, but decreased the n-6/n-3 ratio by decreasing the abundance of bacteria that hydrogenate C18:3n3 and increasing the abundance of bacteria that hydrogenate C18:2n6.

Distinct blood and milk 18-carbon fatty acid proportions and buccal bacterial populations in dairy cows differing in reticulorumen pH response to dietary supplementation of rapidly fermentable carbohydrates.

Nine Holstein dairy cows were fed diets with increasing proportions of rapidly fermentable carbohydrates (RFCH) to investigate the effect on reticular pH, milk fat content (MFC), 18-carbon fatty acid proportions in blood plasma and milk, and bacterial community in buccal swab samples. Inter-animal variation was expected in terms of reticular pH response upon higher RFCH proportions, which would be reflected in the occurrence or not of milk fat depression (MFD). Moreover, this variation in occurrence of MFD was hypothesized to be related to differences in blood and milk fatty acid proportions and in the bacterial community in buccal samples. Cows were fed a total mixed ration throughout the experiment, which consisted of 4 periods: adaptation (d 0-4) and low (d 5-18), increasing (d 19-24), and high RFCH (d 25-28). During the increasing RFCH period, the standard concentrate (211 g of starch/kg of dry matter) was gradually and partly replaced by a concentrate high in RFCH (486 g of starch/kg of dry matter). The reticular pH was measured using a bolus and the time below pH 6.00 was calculated on a daily basis. On d 13, 14, 25, 27, and 28, plasma and milk samples were collected and analyzed for 18-carbon fatty acid proportions, and buccal swabs were collected for bacterial community analysis based on 16S rRNA gene amplicon sequencing. Inter-animal variation was observed in terms of reticular pH, which allowed us to divide the cows into 2 groups: tolerant (time below pH 6.00 ≤ 0.1 h/d) and susceptible cows (time below pH 6.00 ≥ 1.26 h/d). The lower reticular pH of susceptible cows was accompanied by lower MFC. Both groups already differed in reticular pH and MFC during the low-RFCH period. Furthermore, higher RFCH amounts did not decrease the reticular pH in either of the 2 groups. Nevertheless, MFD was observed in both groups during the high-RFCH period compared with the low-RFCH period. Lower MFC in animals with lower reticular pH or during the high-RFCH period was associated with a shift in 18-carbon fatty acids toward trans-10 at the expense of trans-11 intermediates, which was observed in plasma as well as in milk samples. Moreover, lower MFC was accompanied by shifts in the relative abundance of specific bacteria in buccal samples. Genera Dialister, Sharpea, Carnobacterium, Acidaminococcus, and uncultured genera belonging to the Betaproteobacteria were more abundant in situations with greater trans-10 proportions.