Symposium review: The importance of the ruminal epithelial barrier for a healthy and productive cow

The stratified squamous ruminal epithelium is the main site for absorption of key nutrients (e.g., short-chain fatty acids; SCFA) and electrolytes (e.g., sodium and magnesium). The absorptive function has to be highly selective to prevent simultaneous entry of microbes and toxins from the rumen into the blood. As such, epithelial absorption is primarily transcellular, whereas the paracellular pathway appears rather tightly sealed. A network of tight junction (claudin-1, claudin-4, and occludin) and tight junction-associated proteins (e.g., zonula occludens) accomplishes the latter. When microbial fermentation activity is high such as with highly fermentable diets, rumen epithelial functions are often challenged by acidity, high osmolarity, toxins (e.g., endotoxin and histamine), and immune mediators (inflammatory mediators and cytokines) released during local and systemic inflammation. Epithelial damage by low pH in combination with high luminal SCFA concentrations is not immediately reversible and may initially aggravate upon return to physiological pH. In contrast, barrier opening upon hyperosmolarity is acutely transient. The initial insults set by luminal acidity and SCFA and the increasing concentrations of microbial-associated molecular patterns such as lipopolysaccharides are key factors that trigger inflammation not only in the rumen but also in the hindgut (cecum and colon), which reach out to the liver and other organs, causing systemic inflammation. Low feed intake during parturition, transportation, heat stress, or disease is the second most relevant challenge for the ruminal epithelial barrier. The barrier opening is usually only transient and quickly restored upon refeeding. Due to a rapid, dose-dependent, and prolonged decrease in absorption capacity for SCFA, however, any feed restriction increases the odds for postrestriction subacute ruminal acidosis. Inflammation due to acidosis can be alleviated by supplemental thiamine, yeasts, and plant bioactive (phytogenic) compounds. Butyrate is used in weaning calves to support ruminal barrier development; however, excess butyrate may promote hyperkeratosis, parakeratosis, and epithelial injury in the fully developed rumen of adult cows. Further research is needed to enhance the understanding of the various factors that counteract barrier impairment and help barrier restoration during acidogenic feeding, especially when concurring with unavoidable periods of feed restriction.     

Evaluation of the response of ruminal fermentation and activities of nonstarch polysaccharide-degrading enzymes to particle length of corn silage in dairy cows

The main objective of this study was to evaluate effects of particle length (PL) of corn silage (CS) on distribution of dietary particle fractions, contents of physically effective neutral detergent fiber (peNDF), cows’ intake patterns and sorting activity, fermentation pro-file, and activities of nonstarch polysaccharide-degrading enzymes as well as degradation in the rumen and total tract in lactating dairy cows. Four ruminally cannulated Holstein cows, weighing 624 ± 50 kg and 60 ± 8 d in milk, were fed ad libitum 3 total mixed rations [about 16% crude protein, 34% neutral detergent fiber (NDF), and 7 MJ of net energy of lactation/kg of dry matter (DM)] containing on DM basis 50% concentrate, 10% grass hay, and 40% CS with 3 different theoretical PL at harvesting (14, 8.1, and 5.5 mm for long, medium, and short, respectively). Results showed that the amount of DM retained on sieves with 19- and 8-mm screens of Penn State Particle Separator decreased linearly with decreasing PL of CS. The latter was reflected in a significant decrease in the content of dietary peNDF including both the DM (peNDF_>8) and the NDF (peNDF_>8-NDF) retained on 19- and 8-mm screens. In contrast, the fraction of particles retained between the 1.18- and 8-mm screens was increased, such that no differences among the diets were observed regarding the content of peNDF that includes DM of particles >1.18 mm (peNDF_>1.18). The intake of particles retained between the 1.18- and 8-mm screens increased linearly, whereas the intake of peNDF_>1.18 increased quadratically with decreasing PL of CS. Sorting consumption was reduced by feeding the short CS, which was reflected in a reduced proportion of propionate and increased acetate-to-propionate ratio and butyrate pro-portion in the rumen. In contrast, no effects of PL of CS were observed on the concentration of total volatile fatty acids and pH in the rumen. In general, decreasing the PL of CS significantly increased the activities of nonstarch polysaccharide-degrading enzymes. However, greater ruminal and total tract degradation of fiber and nonfiber carbohydrates were observed only by medium CS. Results of the present study suggest that in addition to fractions of long particles (i.e., >8 mm), the particle fraction retained between 1.18- and 8-mm should also be considered to better predict rumen conditions and digestion. In conclusion, a moderate reduction of PL of CS has beneficial effects on nutrient digestion, and may maximize feed efficiency and energy supply in high-yielding dairy cows.     

Feeding rolled barley grain steeped in lactic acid modulated energy status and innate immunity in dairy cows

Feeding dairy cows large proportions of cereal grain is commonly associated with rumen acidosis, activation of innate immunity, and perturbation of intermediary metabolism. We previously showed that steeping barley grain in 0.5% lactic acid (LA) decreased the rate of starch degradation, lowered the risk of subacute rumen acidosis, modulated rumen fermentation profile, and increased milk fat content in dairy cows. This study sought to investigate whether feeding of LA-treated barley grain would affect carbohydrate and lipid metabolism as well as innate immunity. Eight rumen-fistulated late-lactation (approximately 217 d in milk, DIM) Holstein cows were randomly assigned, in a 2 × 2 crossover design, to 1 of the 2 dietary treatments consisting of 27% (dry matter basis) rolled barley grain steeped for 48 h in an equal volume (wt/vol) of tap water (CTR) or 0.5% LA (TRT). Each experimental period lasted 21 d, with the first 11 d for diet adaptation. Blood and rumen samples were collected on d 12, 15, 17, and 21 of the experimental period before the morning feeding to evaluate the effects of dietary treatment on preprandial day-to-day variation of plasma and rumen variables. To establish the effect of treatment on diurnal variation of plasma variables, blood samples were collected on the last day of each period at 0, 2, 4, 6, 8, 10, and 12 h after the morning feeding (i.e., 0800 h). Results of the day-to-day study showed that cows fed the TRT diet had greater overall preprandial concentrations of glucose, cholesterol, and insulin, and a lower concentration of haptoglobin in plasma. Diurnal data indicated lower concentrations of haptoglobin and serum amyloid A and a tendency for greater plasma lactate in cows fed the TRT diet. A treatment by time interaction was observed for glucose, lactate, insulin, haptoglobin, and lipopolysaccharide-binding protein, suggesting a role for both the processing of grain and the time of sampling on those variables. No effect of diet on plasma concentrations of cortisol, β-hydroxybutyrate, and nonesterified fatty acids or rumen endotoxin was evidenced. Taken together, our results demonstrated that feeding barley grain steeped in 0.5% LA modulated both energy status and innate immunity of dairy cows fed relatively high levels (45% of dry matter) of dietary concentrate.     

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.     

Feeding barley grain steeped in lactic acid modulates rumen fermentation patterns and increases milk fat content in dairy cows

The objectives of the present in vivo and in situ trials were to evaluate whether feeding barley grain steeped in lactic acid (LA) would affect rumen fermentation patterns, in situ dry matter (DM) degradation kinetics, and milk production and composition in lactating dairy cows. The in vivo trial involved 8 rumen-fistulated Holstein cows fed once daily a total mixed ration containing rolled barley grain (27% in DM) steeped for 48 h in an equal quantity of tap water (CTR) or in 0.5% LA (TRT) in a 2 × 2 crossover design. The in situ trials consisted of incubation of untreated rolled barley grain in cows fed CTR or TRT diets and of incubation of 3 different substrates including CTR or barley grain steeped in 0.5% or 1.0% LA (TRT1 and TRT2, respectively) up to 72 h in the rumen. Results of the in vivo trial indicated that cows fed the TRT diet had greater rumen pH during most intensive fermentation phases at 10 and 12 h post-feeding. The latter effect was associated with a shorter duration in which rumen pH was below 5.8 for cows fed the TRT diet (2.4 h) compared with CTR diet (3.9 h). Furthermore, cows fed the TRT diet had lower concentrations of volatile fatty acids at 2 and 4 h post-feeding. In addition, concentrations of preprandial volatile fatty acids were lower in the rumen fluid of cows fed the TRT diet. Results also showed that molar proportion of acetate was lower, whereas propionate tended to increase by feeding cows the TRT diet. Cows fed the TRT diet demonstrated greater rumen in situ lag time of substrate DM degradation and a tendency to lower the fractional degradation rate. Other in situ results indicated a quadratic effect of LA on the effective rumen degradability of substrates whereby the latter variable was decreased from CTR to TRT1 but increased for TRT2 substrate. Although the diet did not affect actual milk yield, fat-corrected milk, percentages of milk protein, and lactose and concentration of milk urea nitrogen, cows fed the TRT diet increased milk fat content and tended to increase fat:protein ratio in the milk. In conclusion, results demonstrated that treatment of barley grain with LA lowered the risk of subacute rumen acidosis and maintained high milk fat content in late-lactating Holstein cows fed diets based on barley grain.     

Relationships between rumen lipopolysaccharide and mediators of inflammatory response with milk fat production and efficiency in dairy cows

The main objective of this study was to evaluate correlative relationships between rumen lipopolysaccharide (LPS) and mediators of acute phase response with milk fat yield and efficiency in dairy cows challenged with graded amounts of barley grain in the diet. An additional aim of the study was to quantify the intercow variation in relation to milk fat production and acute phase response in cows fed graded amounts of grain. Eight primiparous, lactating Holstein cows (60 d in milk) were assigned to 1 of the 4 total mixed rations containing barley grain at 0, 15, 30, and 45% (dry matter basis) in a replicated 4 × 4 Latin square design. Free rumen LPS, plasma acute phase proteins, and milk fat content were quantified in multiple samples collected on d 5 and 7 of the measurement periods shortly before the morning feeding. Results showed markedly greater concentrations of rumen LPS with increasing dietary grain level. The correlative analysis revealed strong negative relationships between rumen LPS and milk fat content and yield. The predictor variable of rumen LPS explained 69% of the variation during the milk fat reduction of the cows. The stronger depression in milk fat percentage was obtained when rumen LPS exceeded a threshold of 5,564 ng/mL, corresponding to a milk fat content of 3.39%. The increase in concentration of rumen LPS was also associated with declines in milk fat yield and 3.5% fat-corrected milk (R² = 0.50), as well as milk energy efficiency (R² = 0.43). The correlative analysis also indicated that the increase of plasma C-reactive protein (CRP) in response to higher grain feeding was associated with a linear decrease of milk fat content and yield (R² = 0.28 to 0.46). Furthermore, the statistical analysis revealed high percentages of intercow variation related to milk fat variables, as well as the responses of rumen LPS and plasma CRP. Taken together, the current results implicate rumen LPS and the host CRP response in the lowering of milk fat content and milk energy efficiency in dairy cows fed high-grain diets. Further research is warranted to understand the mechanism(s) by which rumen LPS and inflammatory responses to LPS lower milk fat synthesis and milk energy efficiency and to develop novel strategies for their prevention.     

Models to predict the risk of subacute ruminal acidosis in dairy cows based on dietary and cow factors: A meta-analysis

The present research aimed at developing practical and feasible models to optimize feeding adequacy to maintain desired rumen pH conditions and prevent subacute ruminal acidosis (SARA) in dairy cows. We conducted 2 meta-analyses, one using data from recent published literatures (study 1) to investigate the prediction of SARA based on nutrient components and dietary physical and chemical characteristics, and another using internal data of our 5 different published experiments (study 2) to obtain adjustments based on cow status. The results of study 1 revealed that physically effective neutral detergent fiber inclusive of particles >8 mm (peNDF >8) and dietary starch [% of dry matter (DM)] were sufficient for predicting daily mean ruminal pH {y = 5.960 – (0.00781 × starch) + (0.03743 × peNDF >8) – [0.00061 × (peNDF >8 × peNDF >8)]}. The model for time of pH suppression (<5.8 for ruminal pH or <6.0 for reticular pH, min/d) can be predicted with additionally including DMI (kg/d): 124.7 + (1.7007 × DMI) + (20.9270 × starch) + (0.2959 × peNDF >8) – [0.0437 × (DMI × starch × peNDF >8)]. As a rule of thumb, when taken separately, we propose 15 to 18% peNDF >8 as a safe range for diet formulation to prevent SARA, when starch or NFC levels are within 20 to 25% and 35 to 40% ranges, respectively. At dietary starch content below 20% of DM, grain type was insignificant in affecting ruminal pH. However, increasing dietary starch contents by using corn as the sole grain source could lead to more severe drops of pH compared with using grain mix based on barley and wheat, as underlined by an interaction between starch content and grain type. Data from study 2 emphasized an increased risk of SARA for cows in the first and second lactation with lower mean pH (0.2 units) and double amounts of time at pH <5.8 compared with the cows with ≥3 parities. Given that a lower ruminal pH is expected in these high-risk cows, it is advisable to keep the lower end of recommended starch (20%) and higher peNDF >8 (18%) contents in the diet of these cows. Overall, the present study underlines the possibility of predicting SARA based on dietary factors including peNDF >8 and starch contents, as well as DMI of the cows, which can be practically implemented for optimal diet formulation for dairy cows. With more data available, future studies should attempt to improve the predictions by including additional key dietary and cow factors in the models.