Dietary carbohydrate sources differently prime the microbial ecosystem but not the epithelial gene expression profile along the complete gut of young calves.

BACKGROUND: Recent data indicated similar growth performance of young calves fed solely high-quality hay instead of a starter diet based on starchy ingredients. Yet, providing exclusively such distinct carbohydrate sources during early life might specifically prime the microbiota and gene expression along the gut of young calves, which remains to be explored. We investigated the effects of starter diets differing in carbohydrate composition, that is medium- or high-quality hay and without or with 70% concentrate supplementation (on fresh matter basis), across the gastrointestinal tract (GIT) of weaned Holstein calves (100 ± 4 days of age) using 16 S rRNA gene sequencing and analyses of short-chain fatty acids and host epithelial gene expressions. RESULTS: The concentrate supplementation drastically decreased microbial diversity throughout the gut, which was also true to a much lesser extent for high-quality hay when compared to medium-quality hay in the foregut. Similarly, the factor concentrate strongly shaped the diet-associated common core microbiota, which was substantially more uniform along the gut with concentrate supplementation. The fermentation profile shifted towards less acetate but more propionate with concentrate supplementation in almost all gut sections, corresponding with higher abundances of starch-utilizing bacteria, while major fibrolytic clusters declined. Noteworthy, the n-butyrate proportion decreased in the rumen and increased in the colon with concentrate, showing an opposite, gut site-dependent effect. Both dietary factors modestly influenced the host epithelial gene expression. CONCLUSIONS: Concentrate supplementation clearly primed the microbial ecosystem on a starch-targeted fermentation with characteristic genera occupying this niche along the entire GIT of calves, whereas the microbial differentiation due to hay quality was less distinct. Overall, changes in the microbial ecosystem were only marginally reflected in the targeted transcriptional profile of the host epithelium.

Detection of Hypoglycin A and MCPrG Metabolites in the Milk and Urine of Pasture Dairy Cows after Intake of Sycamore Seedlings.

Hypoglycin A (HGA), methylenecyclopropylglycine (MCPrG), hypoglycin B (HGB), and γ-glutamyl-α-(methylenecyclopropyl) glycine (γ-glutamyl-MCPrG) are secondary plant metabolites occurring in sycamore maple (Acer pseudoplatanus) as well as several other Sapindaceae (e.g., Blighia sapida). By interfering with energy metabolism, they may cause severe intoxication in humans and other species. However, to date, there is not enough data available concerning the intake, metabolism, or excretion of sycamore maple toxins in dairy cows. In May 2022, five cows were observed over four days, when they had first access to a pasture with two sycamore maples. Grazing of their seedlings that grew numerously in between the pasture plants was monitored by direct observation. Milk samples were drawn both from individual cows and from the bulk tank. Spontaneous urine samples were collected from all cows on day 3 after access to the pasture. Seedlings (100 g) were sampled on the pasture and analyzed, together with milk and urine samples, for sycamore toxins and their metabolites using liquid chromatography-tandem mass spectrometry and liquid chromatography-high-resolution mass spectrometry. Cows ingested sycamore seedlings while grazing. Values of HGA in milk were below the limit of quantification. However, metabolites of HGA and MCPrG were detected in individual milk samples already at the end of the first day of grazing. Urine samples of all five cows showed higher concentrations of conjugated HGA and MCPrG metabolites than in milk. Observations suggest that dairy cows may have a low susceptibility toward sycamore maple toxins. However, whether this could be attributed to foregut fermenting species in general requires further elucidation.

Feeding concentrate with hay of different qualities modulates rumen histology, development of digestive tract as well as slaughter performance and meat quality of young dairy calves.

Concentrate-rich starter diets are commonly fed to dairy calves to stimulate growth performance. However, feeding high amounts of starter feed with low inclusion of forage fibre may jeopardise the development of the gastrointestinal tract (GIT). Moreover, studies investigating the effects of feeding on carcass and meat quality of young calves at rearing are rare. The objective of this research was to investigate the effect of hay quality and concentrate inclusion on the traits of GIT development, slaughter performance and veal quality of young dairy calves. The feeding trial covered the first 14 weeks of life. Seventeen male and three female Holstein calves (n = 20) were randomly allocated to four experimental groups, which received besides acidified whole milk different solid feeds: (1) 100% medium-quality hay (MQH), (2) 100% high-quality hay (HQH), (3) 30% medium-quality hay and 70% concentrate (MQH+C) and (4) 30% high-quality hay and 70% concentrate (HQH+C). The acidified whole milk was fed in the first 12 weeks of life, and calves had ad libitum access to solid feed and water from birth till slaughter. Calves were kept in individual boxes equipped with straw and slaughtered at the end of week 14. After slaughter, gut development traits, rumen histology, slaughter performance and meat quality were assessed. Overall, both concentrate inclusion and hay quality showed major effects on rumen histology and development of the GIT in dairy calves with minimal effects on most carcass cuts and meat quality traits. Concentrate-fed calves had significantly higher average daily gains, final body weights, blood amounts and proportions of organs from the circulatory and respiratory systems. Proportions of liver and kidneys were lowest in MQH-fed calves. The proportion of GIT was significantly lower in groups fed concentrates, but the weight of the reticulorumen was unaffected by solid feed. Concentrate feeding led to thicker keratin layer and epithelium as well as wider papillae in the rumen. Hay quality particularly affected the width of the papilla and epithelium thickness, while feeding hay without concentrate enhanced the thickness of submucosa and muscularis, as well as the size of parotid glands. In conclusion, the type of solid feed affects the development of the GIT with concentrate feeding holding the risk to induce keratinisation of rumen epithelium while enhancing performance and carcass traits.

Investigations on the Transfer of Quinolizidine Alkaloids from Lupinus angustifolius into the Milk of Dairy Cows.

Lupin varieties with a low content of quinolizidine alkaloids (QAs) like blue sweet lupin (BSL) have long been used as a protein source for dairy cows. A health concern for humans may arise from the transfer of acute toxic QAs from feed into cow's milk. This study is the first to quantify the transfer of QAs from BSL into cow's milk with experimental and modeling methods. Four lactating dairy cows were subjected to two 7 day feeding periods with 1 and 2 kg/d BSL, respectively, each followed by a depuration period. BSL contained 1774 mg/kg dry matter total QAs. Individual milk samples were taken twice daily and QA contents in feed and milk determined with liquid chromatography-tandem mass spectrometry. Transfer of QAs into the milk was already seen with the administration of 1 kg/d BSL, with differences in transfer rates (TRs) between individual QAs. A toxicokinetic model was derived to quantify and predict QA feed-to-food transfer. For the four most prominent QAs, our model shows an α-half-life of around 0.27 d. TRs were obtained for six QAs and were between 0.13 (sparteine) and 3.74% (multiflorine). A toxicological assessment of milk containing QAs as measured in this study indicated a potential health concern.

Rumen Metabolism of Senecio Pyrrolizidine Alkaloids May Explain Why Cattle Tolerate Higher Doses Than Monogastric Species.

Rumen metabolism of Senecio pyrrolizidine alkaloids (PAs) and their N-oxide forms was studied by mass spectrometry in in vitro batch culture incubates and confirmed in in vivo samples. Most N-oxides were found to undergo rapid conversion to their corresponding free bases, followed by biotransformation to metabolites hydrogenated at both the necine base and the necic acid moiety. Therefore, rumen metabolism can be considered a detoxification step, as saturated necine base structures are known as the platyphylline type, which is regarded as less or nontoxic. Individual Senecio PAs, such as jacoline, are metabolized slowly during rumen fermentation. PAs that showed limited biotransformation in the rumen in this study also showed limited transformation and CYP-mediated bioactivation in the liver in other studies. This could not only explain why PAs that are comparatively metabolically stable can pass into milk but also suggest that such PAs might be considered compounds of lesser concern.

Shift of dietary carbohydrate source from milk to various solid feeds reshapes the rumen and fecal microbiome in calves.

The transition from milk to solid diets drastically impacts the gut microbiome of calves. We explored the microbial communities of ruminal fluid and feces of Holstein calves when fed milk on d 7 of life, and when fed solid feeds based on either medium- or high-quality hay with or without concentrate inclusion (70% in fresh matter) on d 91. Ruminal fluid and feces had distinct microbial compositions already on d 7, showing that niche specialization in early-life gut is rather diet-independent. Changes between d 7 and d 91 were accompanied by a general increase in microbial diversity. Solid diets differed largely in their carbohydrate composition, being reflected in major changes on d 91, whereby concentrate inclusion was the main driver for differences among groups and strongly decreased microbial diversity in both matrices. Fecal enterotyping revealed two clusters: concentrate-supplemented animals had an enterotype prevalent in Prevotella, Succinivibrio and Anaerovibrio, whereas the enterotype of animals without concentrate was dominated by fibrolytic Ruminococcaceae. Hay quality also affected microbial composition and, compared to medium-quality, high-quality hay reduced alpha-diversity metrics. Concluding, our study revealed that concentrate inclusion, more than hay quality, dictates the establishment of niche-specific, microbial communities in the rumen and feces of calves.

Effects of ensiling conditions on pyrrolizidine alkaloid degradation in silages mixed with two different Senecio spp.

Pyrrolizidine alkaloid (PA) producing plants like Senecio jacobaea or Senecio vernalis are undesirable in fields for forage production, since PA are toxic to animals and humans. Previous studies have shown that ensiling can decrease the PA content in forages; however, no direct comparison of diverse PA from different Senecio spp. under various ensiling conditions has been made. Therefore, it was hypothesised that individual PA might react differently to ensiling, and silage inoculation with Lactobacillus will affect PA degradation because of a quick drop in pH, contrastingly to poor silage qualities resulting from contamination with soil. Laboratory scale grass silages were prepared in a multifactorial design with two levels of dry matter contents, four ensiling treatments and two storage durations (10 and 90 d). For each combination, four replicates were prepared individually. Ensiling treatments were (1) 10 ml water per kg fresh matter as control (CON), (2) 10 ml heterofermentative Lactobacillus buchneri strain LN4637 at 3 · 105 cfu/kg fresh matter plus 25 g molasses/kg fresh matter (LBHE), (3) 10 ml homofermentative lactobacilli at 3 · 105 cfu/kg fresh matter plus 25 g molasses/kg fresh matter (LBHO) and (4) 10 g soil/kg fresh matter (SOIL). Treatments affected formation of fermentation acids. Acetic acid was highest with treatment LBHE, and butyric acid was highest with treatment SOIL. All ensiling treatments effectively reduced total PA content by degrading the PA N-oxide (PANO) fraction. In parallel, though, the fraction of the tertiary base forms increased by around one-tenth of the original PANO content. Contents of jaconine and senkirkine were higher after ensiling than before, with regards to the sum of PA and PANO for jaconine, indicating higher stability or new formation through degradation of other PA. Overall, ensiling offers opportunities to decrease the PA-PANO content in feed and therefore lowers the risk of intoxication by Senecio in livestock.

A review on the potentials of using feeds rich in water-soluble carbohydrates to enhance rumen health and sustainability of dairy cattle production.

Cows are adapted to degrade structural plant carbohydrates (SC), such as cellulose and hemicelluloses, prevailing in grasses. Yet, the need for energy-dense diets in many intensive dairy production systems has shifted the dairy cattle's diet from SC-rich to high levels of starch. Feeding of starch-rich diets increases the risk of ruminal acidosis in cows, and feeding starch in the form of grains intensifies the competition over cereal grains and arable land among different livestock species, as well as between livestock and humans. Besides cellulose and hemicelluloses, grasses are also often rich in water-soluble carbohydrates (WSC), which comprise mono-, di-, oligo- and polysaccharides (fructans). Although the ruminal fermentation profile of mono- and disaccharides resembles that of starch, the degradation of oligo- and polysaccharides is slower, and their fermentation elicits a rather protecting effect on ruminal pH. When harvested in an early phase (i.e. ear emergence), grass hay and silages can reach WSC levels up to 150-200 g kg-1 dry matter and energy levels close to starch-rich diets, allowing a significantly reduced inclusion of concentrate supplements. By doing so, this will enhance both rumen health and the sustainability of milk production. However, because the WSC are chemically very heterogeneous, the patterns and extent of their ruminal fermentation are difficult to predict without a clear analytical characterization. This review article aims to summarize both the benefits and potentials, as well as the challenges, with respect to using WSC-rich feedstuffs in the nutrition of dairy cattle and their effects on ruminal fermentation characteristics and milk production. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Predicting the transfer of contaminants in ruminants by models - potentials and challenges.

Undesirable substances in feed can transfer into foods of animal origin after ingestion by livestock animals. These contaminants in food may threaten consumer health. Commonly, feeding trials are conducted with animals to assess the transfer of undesirable substances into animal tissues or milk. Such feeding trials explore the effects of the various physiological systems (e.g., ruminant and non-ruminant gastro-intestinal tracts) as well as different livestock production intensities on transfer. Using alternative methods to mimic the complex physiological processes of several organs is highly challenging. This review proposes a potential cascade of in vitro and ex vivo models to investigate the transfer of contam­inants from feed into foods of animal origin. One distinct challenge regarding the models for ruminants is the simulation of the forestomach system, with the rumen as the anaerobic fermentation chamber and its epithelial surfaces for absorption. Therefore, emphasis is placed on in vitro systems simulating the rumen with its microbial ecosystem as well as on ex vivo systems to replicate epithelial absorption. Further, the transfer from blood into milk must be evaluated by employing a suitable model. Finally, in silico approaches are introduced that can fill knowledge gaps or substitute in vitro and ex vivo models. Physiologically-based toxicokinetics combines the information gained from all alternative methods to simulate the transfer of ingested undesirable substances into foods of animal origin.

Stability of pyrrolizidine alkaloids from Senecio vernalis in grass silage under different ensilage conditions.

BACKGROUND: This study evaluated the degradation of pyrrolizidine alkaloids (PAs) from eastern groundsel (Senecio vernalis) in grass silage prepared with different inoculants. Silages were produced from ryegrass with 230 g kg-1 dry matter (DM) content and mixed with eastern groundsel (9:1; w/w fresh matter basis) containing 5.5 g kg-1 DM PA. Treatments were: CON (untreated control), LP (3.0 × 105 cfu g-1 Lactobacillus plantarum DSMZ 8862/8866) or LBLC (7.3 × 104 cfu g-1 Lactobacillus buchneri LN40177 / Lactobacillus casei LC32909), and each of the treatments in combination with 30 g kg-1 molasses. Silages were prepared in glass jars and opened after 3, 10, and 90 days. Fermentation characteristics were determined and the PAs analyzed. RESULTS: Although the levels of fermentation acids differed between treatments, results indicated good quality of all silages during 90 days. Significant time (P < 0.001) and treatment (P < 0.001) effects were observed for PAs. Concentrations of senecionine and seneciphylline decreased with molasses, declined over time, and were negatively correlated with lactic, propionic, and butyric acid, or with lactic and butyric acid in case of seneciphylline. In all silages, seneciphylline and senecionine N-oxides were undetectable after 3 days, whereas senkirkine, the most abundant PA, remained stable. CONCLUSIONS: Silage prepared from grass contaminated with eastern groundsel still contained high PA levels, and was hence a potential health hazard. Molasses supplementation reduced concentrations of senecionine and seneciphylline, while the bacterial inoculants had no effect. Other potentially toxic PA metabolites were not analyzed in the present study and further research is needed. © 2019 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Graded replacement of corn grain with molassed sugar beet pulp modulates the fecal microbial community and hindgut fermentation profile in lactating dairy cows.

High starch lactation diets not only enhance the risk of subacute ruminal acidosis but also of hindgut acidosis, which increases the risk of dysbiosis and the depression of fiber degradation. We recently showed that replacing corn with molassed sugar beet pulp (Bp) improved fiber degradation in high-producing dairy cattle, possibly because of an improvement of rumen and hindgut conditions for microbes by Bp feeding. However, little is known about the effects of high inclusion rates of Bp on hindgut microbes and fermentation. Thus fecal grab samples were taken from 18 high-yielding Simmental cows after 28 d of feeding 3 different levels of Bp (n = 6) for bacterial 16S rRNA amplicon sequencing. In addition, the reticular pH was continuously monitored with indwelling sensors and eating and ruminating behavior was evaluated with noseband sensors. The Bp inclusion rates were 0 g/kg (i.e., no Bp inclusion as control, CON), 120 g/kg (12Bp), or 240 g/kg (24Bp) replacing corn grain and limestone on a dry matter basis. The amount of time spent eating and ruminating was unaffected by Bp level, and the daily fluctuation in the reticular pH was reduced by 25% with Bp inclusion from 0.8 in the CON diet to 0.6 in 24Bp fed animals. Also, the fecal pH tended to increase with dietary Bp inclusion. Fecal acetate production showed a quadratic tendency with the lowest concentration (58.9%) of the total short-chain fatty acid in the 12Bp treatment. Inclusion of Bp up to 24% of the diet decreased the fecal butyrate proportion by 27%. The Shannon diversity index was increased from 5.50 to 8.09 with dietary Bp inclusion indicating increased species diversity. Of the 200 most abundant operational taxonomic units, 25 were increased by dietary Bp inclusion, whereas 15 were decreased and 7 were quadratically affected. The second most abundant group was proposed taxon "CF231" of the family Paraprevotellaceae. Although it accounted for only 2.52% of the operational taxonomic units in the CON diet, it was increased by 64% with dietary Bp inclusion. The largest relative change in the abundance was found for the genus Fibrobacter that increased more than 14-fold from 0.04% (CON) to 0.66% (24Bp). In conclusion, feeding molassed sugar beet pulp as partial substitution of corn up to 240 g/kg is a viable alternative that promotes ruminal and hindgut fermentation by supporting physiological pH and bacterial diversity.

Effects of the supplementation of plant-based formulations on microbial fermentation and predicted metabolic function in vitro.

This study aimed at testing the effects of three different formulations of feed supplements based on three different combinations of plant derived alkaloids, prebiotics, tannins, vitamins and minerals on rumen fermentation and the microbiome in vitro. A Rusitec experiment was conducted in 2 identical runs using a complete randomized design with 3 replicates per treatment resulting in total of 6 treatment combinations (n = 6). Each run lasted 12 d with sampling occurring in the last 5 d. Diets were a standard dairy ration (60:40; concentrate:forage) supplemented with one of 3 different plant-based combinations (PI, PII, and PIII) at a level of 100 mg/l and a non-supplemented control (basal diet, control). Microbial DNA samples were taken on the last day of each run and the 16S rRNA target gene sequenced using Illumina MiSeq technology. The supplementations had no effect on the pH, methane and carbon dioxide production. However, both total SCFA (P = 0.08) and molar concentrations of acetate (P = 0.06) tended to be increased in the treatment groups in comparison to control, with PII having the highest overall values (102.7 mmol/L and 43.3 mmol/L, respectively). Alpha diversity indices Shannon, Simpson and Chao1 showed no effect of supplementations or combinations. The addition of PII increased the relative abundance of Bacteroidetes compared to all other treatments (P = 0.05). Supplementation with plant-based combinations reduced the relative abundance of Pyramidobacter from the family Dethiosulfovibrionaceae in comparison with the control diet (P = 0.05). Evaluation of predicted gene function through PICRUSt analysis showed variation in predicted cellular function and metabolism between bacterial communities supplemented with plant-based combinations compared to the control diet. This shows that the addition of plant-based combinations can have the potential to modulate the metabolic function of rumen microbes, and likely the production of small-sized rumen metabolites, without disrupting the rumen microbial community structure and diversity.

A nutritional and rumen ecological evaluation of the biorefinery by-product alfalfa silage cake supplemented with Scrophularia striata extract using the rumen simulation technique.

BACKGROUND: By-products of the food production chain are gaining importance as feedstuffs for ruminants. Alfalfa silage cake (AC) is an unexploited biorefinery by-product rich in fiber. The aim of this study was to test AC, using an in vitro rumen simulation technique (Rusitec), for its suitability as a fiber source for cattle. Three diets with similar crude protein (CP) content were formulated; they contained the biorefinery by-product AC, the original alfalfa silage (OA), or a fiber-rich hay. As fibrous feedstuffs are known to promote ruminal methanogenesis, we additionally tested a plant extract of Scrophularia striata (60 mg g-1 dry matter) for its methane mitigation and antimicrobial properties. RESULTS: Diets containing AC displayed lower nutrient degradability, with the largest difference in CP degradation (P < 0.001). Sequencing of microbial DNA revealed several effects of the diet and of the addition of S. striata extract, but no inhibitory effect on methanogens. Likewise, methane production, which, in general, is lower with AC and OA diets, was not inhibited by S. striata extract, while the short chain fatty acid (SCFA) profiles were unaffected. CONCLUSION: Although CP degradation of the AC diet was lower, degradation of the fiber fractions was similar among diets. According to the present results, AC can be used as fibrous feedstuff for ruminants. Supplementation with S. striata extract did not inhibit methane formation. © 2019 Society of Chemical Industry.

Feeding hay rich in water-soluble carbohydrates improves ruminal pH without affecting rumination and systemic health in early lactation dairy cows.

Forages rich in water-soluble carbohydrates (WSC) might be an ideal energy source during early lactation, as they provide both energy for milk production and structural fibre to promote chewing and rumen buffering. Thus, the aim was to investigate feeding strategies based on high-quality hay rich in WSC with graded amounts of concentrate on ruminating behaviour, ruminal pH and systemic health variables. Twenty-four Simmental cows were randomly allocated to four groups beginning 10 days before until 28 days after calving. Diets were 60LQH (60% fibre-rich hay plus 40% concentrate), 60HQH, 75HQH and 100HQH (60%, 75% and 100% high-quality hay, plus 40%, 25% and 0% concentrate, respectively). Hay qualities differed in contents of WSC (110 g vs. 198 g of dry matter [DM]), neutral detergent fibre (646 g vs. 423 g of DM) and crude protein (65 g vs. 223 g of DM). Rumination was recorded using the Rumiwatch system over 4 days during the last week. Weekly serum samples were analysed for the liver enzymes aspartate aminotransferase, glutamate dehydrogenase and γ-glutamyltransferase and the acute phase proteins serum amyloid A and haptoglobin. Four cows per group received a wireless pH sensor orally placed into the rumen one week before the expected calving date. Daily time spent chewing did not differ between groups. Likewise differences in minimum, maximum and mean pH-values were not significant, but daily time of reticular pH <6 was longer in cows fed 60LQH compared to cows fed 100HQH (p = 0.043) and in tendency to cows fed 75HQH or 60HQH (p = 0.072 or p = 0.086, respectively). Blood parameters were unaffected by diet. Accordingly the present results demonstrate that feeding hay rich in WSC helped stabilizing the reticuloruminal pH in early lactation dairy cows, even in combination with 40% concentrates in DM.

Scrophularia striata Extract Supports Rumen Fermentation and Improves Microbial Diversity in vitro Compared to Monensin.

In the search for natural alternatives to antibiotic feed additives, we compared the efficacy of two doses of Scrophularia striata extract [S. striata-Low at 40 and S. striata-High at 80 mg g-1 dry matter (DM)] with monensin (monensin) and a negative control in the modulation of rumen fermentation, methane production and microbial abundance in vitro. Microbes were investigated using qPCR and 16S rRNA targeted sequencing. Data showed that the addition of S. striata increased production of total short chain fatty acids (SCFA) in comparison to both monensin and control (P = 0.04). The addition of S. striata increased acetate production, and increased propionate at the higher dosage (P < 0.001). Supplementation of S. striata lowered methane production (P < 0.001) compared to control but with no effect compared to monensin. Ammonia concentration decreased by 52% (P < 0.001) with S. striata-High supplementation (4.14 mmol L-1) compared to control, which was greater than that of monensin (36%). The diversity of rumen bacteria was reduced (P < 0.001) for monensin and S. striata for both the number of observed OTUs and the Chao1 index. Quantitative analysis of Protozoa showed a decrease in the monensin treatment (P = 0.05) compared to control. Archaea copy numbers decreased equally in both S. striata-High and monensin treatments compared to the control group. Supplementation with S. striata increased relative abundances of Fibrobacteres (P < 0.001) and Planctomycetes (P = 0.001) in comparison to both the control and monensin treatments. Significant negative correlations were observed between the abundances of Bacteroides, Fusobacterium, and Succinivibrio genera and methane (r > -0.71; P ≤ 0.001). The abundance of Fibrobacter genera and total SCFA (r = 0.86), acetate (r = 0.75), and valerate (r = -0.51; P < 0.001) correlated positively. These results suggest that S. striata supplementation at 80 mg g-1 DM inclusion, similar to monensin, supports rumen fermentation, lowers methane and ammonia production. However, S. striata supported rumen fermentation toward higher total SCFA and propionate production, while unlike monensin still supported a diverse rumen microbiome and an increase in cellulolytic bacteria such as Fibrobacter.

Correction: Temporal dynamics of in-situ fiber-adherent bacterial community under ruminal acidotic conditions determined by 16S rRNA gene profiling.

[This corrects the article DOI: 10.1371/journal.pone.0182271.].

Lactic acid treatment of by-products and phosphorus level in the diet modulate bacterial microbiome and the predicted metagenome functions using the rumen simulation technique.

This study used a rumen simulation technique to evaluate the effects of soaking of by-product-rich concentrate (BPC) in 5% lactic acid (LAC; vol/vol) on the rumen microbiota, predicted metagenome, fermentation characteristics, and nutrient degradation without or with supplemented P. The diet was supplemented with 1.6 g of P in the form of monocalcium phosphate per kilogram of dry matter in addition to 284 mg of inorganic P/d per fermentor via artificial saliva. Fermentor fluid was collected for analyses of short-chain fatty acids, fermentation gases, redox potential, and microbiota and feed residues for calculation of nutrient degradation. The microbiota composition was assessed using paired-end Illumina (Illumina Inc., San Diego, CA) MiSeq sequencing of the V3 to V5 region of the 16S rRNA gene. Soaking in LAC reduced the contents of crude protein, neutral and acid detergent fibers, and organic matter fractions as well as ash and P content of the BPC. Both the LAC treatment of BPC and the inorganic P modified the relative bacterial abundances mainly within the predominant orders Bacteroidales and Clostridiales. Supervised DIABLO N-integration networking supported that operational taxonomic units related to BS11, Ruminococcaceae, Christensenellaceae, Eubacterium, and Selenomonas were the most discriminant for the LAC-treated BPC, whereas other operational taxonomic units related to BS11, RFN20, Ruminococcus, and Succiniclasticum were best correlated with the inorganic P supplementation. Integration networking also showed that carbohydrate and pyruvate metabolism, biosynthesis of unsaturated fatty acids, and degradation of several xenobiotics were stimulated by the LAC treatment of BPC. Those data supported the enhanced fermentation activity as indicated by increased total short-chain fatty acid concentration, especially propionate and butyrate, and methane, but decreased ruminal crude protein degradation, with the LAC-treated compared with control-treated BPC. In contrast, despite an increased abundance of imputed functions, such as inositol phosphate metabolism, phosphatidylinositol signaling, and fructose and mannose metabolism, the reduced abundance of the imputed Kyoto Encyclopedia of Genes and Genomes pathway "transcription machinery" as well as the decrease in total short-chain fatty acids and nutrient degradation indicated reduced bacterial metabolic activity with the inorganic P supplementation. In conclusion, soaking of BPC in LAC may favor the proliferation of certain fibrolytic bacterial taxa and stimulate their metabolic activity, whereas the supplemented P to a diet already meeting ruminal P needs may impair ruminal nutrient utilization.

Effects of feeding high-quality hay with graded amounts of concentrate on feed intake, performance and blood metabolites of cows in early lactation.

Dairy cows are commonly fed energy-dense diets with high proportions of concentrate feedstuffs to meet the increased energy needs of early lactation. However, feeding large amounts of concentrates may cause rumen acidosis and impact cow health. The hypothesis tested was that the energy supply and metabolic health of early-lactation Simmental cows can be maintained when high-quality hay rich in water-soluble carbohydrates (WSC) and crude protein (CP) is fed, despite the proportion of concentrates in the diet being reduced or even excluded. Twenty-four Simmental cows were allocated to one of four feeding groups beginning 10 d before the expected calving date, until 28 d thereafter. The feeding groups were 60CH (60% conventional fibre-rich hay plus 40% concentrate feed), 60HQH (60% high-quality hay plus 40% concentrate feed), 75HQH (75% high-quality hay plus 25% concentrate feed) and 100HQH (100% high-quality hay). The fibre-rich hay and high-quality hay differed in WSC content (110 g vs. 198 g of dry matter (DM)), neutral detergent fibre (646 g vs. 423 g of DM) and CP (65 g vs. 223 g of DM). Individual feed intake and milk production were monitored daily, and blood samples were collected weekly. Dry matter intake (DMI) and milk yield increased post partum, but 4 weeks post partum, the DMI of cows fed 100HQH only reached a daily mean DMI of 18.6 kg, whereas the DMI of the other groups averaged 21.9 kg (p < 0.046). The negative energy balance was less pronounced in cows fed 75HQH since they showed similar milk yields to the cows fed 60CH and 100HQH, but their energy intake was higher. Concentrations of milk components were similar across rations 60CH, 60HQH and 75HQH, as were most of blood parameters. Cows fed 100HQH responded to the energy deficit post partum with a higher ratio of non-esterified fatty acids to cholesterol and a higher concentration of ß-hydroxybutyrate (significant in comparison to cows fed 75HQH, p < 0.05). In conclusion, feeding high-quality hay with a WSC content of 20% in DM has the potential to decrease the proportion of concentrates in dairy cow feeding in early lactation, but cannot fully replace their supplementation due to a limited rumen capacity for forage intake.

Graded replacement of maize grain with molassed sugar beet pulp modulated ruminal microbial community and fermentation profile in vitro.

BACKGROUND: Molassed sugar beet pulp (Bp) is a viable alternative to grains in cattle nutrition for reducing human edible energy input. Yet little is known about the effects of high inclusion rates of Bp on rumen microbiota. This study used an in vitro approach and the quantitative polymerase chain reaction technique to establish the effects of a graded replacement of maize grain (MG) by Bp on the ruminal microbial community, fermentation profile and nutrient degradation. RESULTS: Six different amounts of Bp (0-400 g kg-1 ), which replaced MG in the diet, were tested using the in vitro semi-continuous rumen simulation technique. The increased inclusion of Bp resulted in greater dietary content and degradation of neutral detergent fibre (P < 0.01). Further, Bp feeding enhanced (P < 0.01) the abundance of genus Prevotella and shifted (P < 0.01) the short-chain fatty acid patterns in favour of acetate and propionate and at the expense of butyrate. A total replacement of MG with Bp resulted in an increased daily methane production (P < 0.01). CONCLUSION: Results suggest positive effects of the replacement of MG by Bp especially in terms of stimulating ruminal acetate and propionate fermentation. However, high replacement rates of Bp resulted in lowered utilization of ammonia and higher ruminal methane production. © 2017 Society of Chemical Industry.

Changes in fibre-adherent and fluid-associated microbial communities and fermentation profiles in the rumen of cattle fed diets differing in hay quality and concentrate amount.

The rumen microbiota enable important metabolic functions to the host cattle. Feeding of starch-rich concentrate feedstuffs to cattle has been demonstrated to increase the risk of metabolic disorders and to significantly alter the rumen microbiome. Thus, alternative feeding strategies like the use of high-quality hay, rich in sugars, as an alternative energy source need to be explored. The aim of this study was to investigate changes in rumen microbial abundances in the liquid and solid-associated fraction of cattle fed two hay qualities differing in sugar content with graded amounts of starchy concentrate feeds using Illumina MiSeq sequencing and quantitative polymerase chain reaction. Operational taxonomic units clustered separately between the liquid and the solid-associated fraction. Phyla in the liquid fraction were identified as mainly Firmicutes, Proteobacteria and Bacteroidetes, whereas main phyla of the fibre-associated fraction were Bacteroidetes, Fibrobacteres and Firmicutes. Significant alterations in the rumen bacterial communities at all taxonomic levels as a result of changing the hay quality and concentrate proportions were observed. Several intermicrobial correlations were found. Genera Ruminobacter and Fibrobacter were significantly suppressed by feeding sugar-rich hay, whereas others such as Selenomonas and Prevotella proliferated. This study extends the knowledge about diet-induced changes in ruminal microbiome of cattle.