J.C. Plaizier

Subacute ruminal acidosis in dairy cows: the physiological causes, incidence and consequences.

During subacute ruminal acidosis (SARA) rumen pH is depressed for several hours per day due to accumulation of volatile fatty acids and insufficient rumen buffering. Surveys suggested an incidence of SARA of between 19% and 26% in early and mid-lactation dairy cows. Causes of SARA include feeding excessive amounts of non-structural carbohydrates and highly fermentable forages, and insufficient dietary coarse fiber. Consequences of SARA include feed intake depression, reduced fiber digestion, milk fat depression, diarrhea, laminitis, liver abscesses, increased production of bacterial endotoxin and inflammation characterized by increases in acute phase proteins. The increase in endotoxin is similar among methods for SARA induction, but depends on the diet fed before induction. Increases in acute phase proteins vary among methods of SARA induction, even when the methods result in similar rumen pH depressions. This suggests that the inflammatory response might not be solely due to bacterial endotoxin in the rumen.

Rumen microbiome composition determined using two nutritional models of subacute ruminal acidosis.

ABSTRACT Subacute ruminal acidosis (SARA) is a metabolic disease in dairy cattle that occurs during early and mid-lactation and has traditionally been characterized by low rumen pH, but lactic acid does not accumulate as in acute lactic acid acidosis. It is hypothesized that factors such as increased gut permeability, bacterial lipopolysaccharides, and inflammatory responses may have a role in the etiology of SARA. However, little is known about the nature of the rumen microbiome during SARA. In this study, we analyzed the microbiome of 64 rumen samples taken from eight lactating Holstein dairy cattle using terminal restriction fragment length polymorphisms (TRFLP) of 16S rRNA genes and real-time PCR. We used rumen samples from two published experiments in which SARA had been induced with either grain or alfalfa pellets. The results of TRFLP analysis indicated that the most predominant shift during SARA was a decline in gram-negative Bacteroidetes organisms. However, the proportion of Bacteroidetes organisms was greater in alfalfa pellet-induced SARA than in mild or severe grain-induced SARA (35.4% versus 26.0% and 16.6%, respectively). This shift was also evident from the real-time PCR data for Prevotella albensis, Prevotella brevis, and Prevotella ruminicola, which are members of the Bacteroidetes. The real-time PCR data also indicated that severe grain-induced SARA was dominated by Streptococcus bovis and Escherichia coli, whereas mild grain-induced SARA was dominated by Megasphaera elsdenii and alfalfa pellet-induced SARA was dominated by P. albensis. Using discriminant analysis, the severity of SARA and degree of inflammation were highly correlated with the abundance of E. coli and not with lipopolysaccharide in the rumen. We thus suspect that E. coli may be a contributing factor in disease onset.

A grain-based subacute ruminal acidosis challenge causes translocation of lipopolysaccharide and triggers inflammation.

The effects of a grain-based subacute ruminal acidosis (SARA) challenge on translocation of lipopolysaccharide (LPS) into the peripheral circulation, acute phase proteins in blood and milk, feed intake, milk production and composition, and blood metabolites were determined in 8 lactating Holstein cows. Between wk 1 and 5 of 2 successive 6-wk periods, cows received a total mixed ration ad libitum with a forage to concentrate (F:C) ratio of 50:50. In wk 6 of both periods, the SARA challenge was conducted by replacing 21% of the dry matter of the total mixed ration with pellets containing 50% wheat and 50% barley. Rumen pH was monitored continuously using indwelling pH probes in 4 rumen cannulated cows. Rumen fluid samples were collected 15 min before feed delivery and at 2, 4, 6, 12, 14, 16, 18, and 24 h after feed delivery for 2 d during wk 5 (control) and wk 6 (SARA). Peripheral blood samples were collected using jugular catheters 15 min before feeding and at 6 and 12 h after feeding at the same days of the rumen fluid collections. The SARA challenge significantly reduced average daily pH from 6.17 to 5.97 and increased the duration of rumen pH below pH 5.6 from 118 to 279 min/d. The challenge reduced dry matter intake (16.5 vs. 19 kg/d), milk yield (28.3 vs. 31.6 kg/d), and milk fat (2.93 vs. 3.30%, 0.85 vs. 0.97 kg/d), and tended to increase milk protein percentage (3.42 vs. 3.29%), without affecting milk protein yield (1.00 vs. 0.98 kg/d). The challenge also increased the concentration of free LPS in rumen fluid from 28,184 to 107,152 endotoxin units (EU)/mL. This was accompanied by an increase in LPS in peripheral blood plasma (0.52 vs. <0.05 EU/mL) with a peak at 12 h after feeding (0.81 EU/mL). Concentrations of the acute phase proteins serum amyloid A, haptoglobin, and LPS-binding protein (LBP) in peripheral blood as well as LBP concentration in milk increased (438.5 vs. 167.4, 475.6 vs. 0, 53.1 vs. 18.2, and 6.94 vs. 3.02 microg/mL, respectively) during SARA. The increase in LBP in combination with the increase in LPS in peripheral blood provides additional evidence of translocation of LPS. Results suggest that the grain-based SARA challenge resulted in translocation of LPS into the peripheral circulation, and that this translocation triggered a systemic inflammatory response.

Alfalfa pellet-induced subacute ruminal acidosis in dairy cows increases bacterial endotoxin in the rumen without causing inflammation

A study was conducted to determine if subacute ruminal acidosis (SARA) induced by feeding alfalfa pellets results in increases in free bacterial lipopolysaccharide (LPS) in rumen fluid and peripheral blood, and acute phase proteins in plasma, and to determine the effect of alfalfa pellet-induced SARA on feed intake, rumen fermentation characteristics, milk production and composition, and blood metabolites. Eight lactating Holstein cows, 4 of which were ruminally cannulated, were used in a 6-wk experiment and were fed once daily at 0900 h. During wk 1, cows received a diet containing 50% of DM as concentrate and 50% of DM chopped alfalfa hay. Between wk 2 and wk 6, alfalfa hay was gradually replaced with alfalfa pellets at the rate of 8% per week to reduce rumen pH. Rumen pH was monitored continuously in the ruminally cannulated cows using indwelling pH probes. Rumen fluid and peripheral blood were sampled 15 min before feed delivery and at 6 h after feed delivery. Based on adopted threshold of SARA of at least 180 min/d below pH 5.6, SARA was induced from wk 3 onwards. Replacing 40% of alfalfa hay with alfalfa pellets quadratically increased the DMI from 18.1 kg/d in wk 1 to 23.4 kg/d in wk 6. This replacement linearly decreased milk yield (32.7 vs. 35.9 kg/d) and milk fat percentage and yield (2.32 vs. 3.22%, and 0.77 vs. 1.14 kg/d, respectively), but increased milk protein percentage and yield (3.80 vs. 3.04%, and 1.23 vs. 1.07 kg/d, respectively). This gradual replacement also linearly increased the daily averages of total volatile fatty acids (90 to 121.9 mM), acetate (53.9 to 66.8 mM), propionate (21.5 to 39.6 mM), and osmolality (277.7 to 293.8 mmol/kg) in the rumen and decreased the acetate to propionate ratio from 2.62 to 1.73. Replacing alfalfa hay with alfalfa pellets linearly increased blood lactate from 1.00 mM in wk 1 to a peak of 3.46 mM in wk 5. Induction of SARA in this study increased free rumen LPS concentration from 42,122 endotoxin unit (EU)/mL in wk 1 to 145,593 EU/mL in wk 6. However, this increase was not accompanied by an increase in LPS (<0.05 EU/mL) and in acute phase proteins serum amyloid-A, haptoglobin, and LPS-binding protein in peripheral circulation. Results suggest that SARA induced by alfalfa pellets increased LPS in the rumen without causing translocation of LPS and an immune response.

Effects of subacute ruminal acidosis challenges on fermentation and endotoxins in the rumen and hindgut of dairy cows

The effects of a grain-based subacute ruminal acidosis (SARA) challenge (GBSC) and an alfalfa-pellet SARA challenge (APSC) on fermentation and endotoxins in the rumen and in the cecum, as well as on endotoxins in peripheral blood, were determined. Six nonlactating Holstein cows with cannulas in the rumen and cecum were used in the study. A 3×3 Latin square arrangement of treatments with 4-wk experimental periods was adopted. During the first 3 wk of each experimental period, all cows received a diet containing 70% forages [dry matter (DM) basis]. In wk 4 of each period, cows received 1 of the following 3 diets: the 70% forage diet fed during wk 1 to 3 (control), a diet in which 34% of the dietary DM was replaced with grain pellets made of 50% ground wheat and 50% ground barely (GBSC), or a diet in which 37% of dietary DM was replaced with pellets of ground alfalfa (APSC). Rumen pH was monitored continuously using indwelling pH probes, and rumen fluid, blood, cecal digesta, and fecal grab samples were collected immediately before feed delivery at 0900 h and at 6 h after feed delivery on d 3 and 5 of wk 4. The time for which rumen pH was below 5.6 was 56.4, 225.2, and 298.8 min/d for the control, APSC, and GBSC treatments, respectively. Compared with the control, SARA challenges resulted in similar reductions in cecal digesta pH, which were 7.07, 6.86, and 6.79 for the control, APSC, and GBSC treatments, respectively. Compared with the control, only GBSC increased starch content in cecal digesta, which averaged 2.8, 2.6, and 7.4% of DM for the control, APSC, and GBSC, respectively. Free lipopolysaccharide endotoxin (LPS) concentration in rumen fluid increased from 10,405 endotoxin units (EU)/mL in the control treatment to 30,715 and 168,391 EU/mL in APSC and GBSC, respectively. Additionally, GBSC increased the LPS concentration from 16,508 to 118,522 EU/g in wet cecal digesta, and from 12,832 to 93,154 EU/g in wet feces. The APSC treatment did not affect LPS concentrations in cecal digesta and feces. All concentrations of LPS in blood plasma were below the detection limit of >0.05 EU/mL of the technique used. Despite the absence of LPS in blood, only GBSC increased the concentration of LPS-binding protein in blood plasma, which averaged, 8.9, 9.5, and 12.1mg/L for the control, APSC, and GBSC treatments, respectively. This suggests that GBSC caused translocation of LPS from the digestive tract but that LPS was detoxified before entering the peripheral blood circulation. The higher LPS concentration in cecal digesta in the GBSC compared with the APSC suggests a higher risk of LPS translocation in the large intestine in GBSC than in APSC.

Effects of Feed Delivery Time on Feed Intake, Milk Production, and Blood Metabolites of Dairy Cows

The objective of the study was to determine the effects of feed delivery time and its interactions with dietary concentrate inclusion and parity on milk production and on 24-h averages and patterns of feed intake and blood metabolites. Four multiparous and 4 primiparous lactating Holstein cows were used in a 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments. Experimental periods included 14 d of adaptation and 7 d of sampling. A higher concentrate diet with a forage:concentrate ratio (dry matter basis) of 38:62 or a lower-concentrate diet with a forage:concentrate ratio of 51:49 was delivered at either 0900 or 2100 h. During sampling periods, daily feed intakes, as well as feed intakes during 3-h intervals relative to feed delivery, were determined. During 2 nonconsecutive days of the sampling period, jugular blood was sampled every 2 h. Average temperature and relative humidity in the experimental facility were 20.4 degrees C and 68.1%, and the maximum daily air temperature did not exceed 25 degrees C. This data does not suggest that cows were heat-stressed. Changing feed delivery time from 0900 to 2100 h increased the amount of feed consumed within 3 h after feeding from 27 to 37% of total daily intake but did not affect daily dry matter intake. The cows fed at 2100 h had lower blood glucose at 2 h after feeding but greater blood lactate and beta-hydroxybutyrate acid at 2 and 4 h after feeding than cows fed at 0900 h. These effects of feed delivery time on the 24-h patterns in blood metabolites may be caused by the greater feed intake during the 3 h after feed delivery of the cows fed at 2100 h. Daily averages of glucose, urea, lactate, and beta-hydroxybutyrate acid and nonesterified fatty acids in peripheral blood were not affected by time of feeding. The change in feed delivery time did not affect milk yield and milk protein but increased milk fat percentage from 2.5 to 2.9% and milk fat yield from 0.98 to 1.20 kg/d in multiparous cows, without affecting milk fat in primiparous cows. The interactions between diet and time of feeding on daily feed intake, milk production, and blood metabolites were not significant. The effects of the time of feed delivery on the 24-h patterns in blood metabolites suggest that this time may affect peripheral nutrient availability. Results of this study suggest beneficial effects of feeding at 2100 h instead of at 0900 h on milk fat production of lactating cows, but parity appears to mediate this effect.

Effects of the chop lengths of alfalfa silage and oat silage on feed intake, milk production, feeding behavior, and rumen fermentation of dairy cows.

Effects of chop length (shorter: 6 mm, or longer: 19 mm) of alfalfa silage and oat silage were determined in 16 mid-lactation Holstein cows, 4 of which were rumen cannulated, using a replicated 4 x 4 Latin square design with a 2 x 2 arrangement of treatments. Experimental periods were 21 d long and consisted of 14 d of adaptation and 7 d of sampling. Cows received a total mixed ration containing [dry matter (DM) basis] 42.0% barley grain-based energy supplement, 10% protein supplement, and 24% of DM longer chop or shorter chop alfalfa silage and 24% of DM longer chop or shorter chop oat silage. Rumen pH was measured continuously, and rumen liquid flow rates were determined in rumen-cannulated cows. Feeding behavior was determined by videotaping, and meal patterns were determined by continuously weighing the feed in the bunk of 8 cows. Reducing the chop length of alfalfa silage and oat silage reduced the average geometric particle length from 14.2 to 10.9 mm and from 13.4 to 10.4 mm, respectively. Reducing the alfalfa chop length did not affect feed intake, whereas reducing the oat silage chop length increased DM intake from to 19.4 to 21.2 kg/d. Reducing the chop lengths of alfalfa silage and oat silage chop length did not affect milk production, rumen fermentation, feeding behavior, meal patterns, and blood metabolites. Daily milk yield, milk fat percentage, and milk protein percentage averaged 36.1 kg/d, 3.00%, and 3.16%, respectively, across diets. The low milk fat percentages suggest that the diets induced subacute ruminal acidosis. This was also substantiated by the rumen pH, which was below 5.6 for more than 122 min/d for all diets. The onset of subacute ruminal acidosis despite apparently adequate dietary neutral detergent fiber content and particle size distribution as well as the long duration of chewing might be attributed to sorting against long feed particles.

Development of ruminal and fecal microbiomes are affected by weaning but not weaning strategy in dairy calves

The nature of weaning, considered the most stressful and significant transition experienced by dairy calves, influences the ability of a calf to adapt to the dramatic dietary shift, and thus, can influence the severity of production losses through the weaning transition. However, the effects of various feeding strategies on the development of rumen or fecal microbiota across weaning are yet to be examined. Here we characterized the pre- and post-weaning ruminal and fecal microbiomes of Holstein dairy calves exposed to two different weaning strategies, gradual (step-down) or abrupt. We describe the shifts toward a mature ruminant state, a transition which is hastened by the introduction of the solid feeds initiating ruminal fermentation. Additionally, we discuss the predicted functional roles of these communities, which also appear to represent that of the mature gastrointestinal system prior to weaning, suggesting functional maturity. This assumed state of readiness also appeared to negate the effects of weaning strategy on ruminal and fecal microbiomes and therefore, we conclude that the shift in gastrointestinal microbiota may not account for the declines in gain and intakes observed in calves during an abrupt weaning.

Population structure of rumen Escherichia coli associated with subacute ruminal acidosis (SARA) in dairy cattle

Previous studies indicated that only subacute ruminal acidosis (SARA), induced by feeding a high-grain diet, is associated with an inflammatory response and increased abundance of Escherichia coli in the rumen. We hypothesized that ruminal E. coli in grain pellet-induced SARA carried virulence factors that potentially contribute to the immune activation during SARA. One hundred twenty-nine E. coli isolates were cultured from the rumens of 8 cows (4 animals per treatment) in which SARA had been nutritionally induced by feeding a high-grain diet (GPI-SARA) or a diet containing alfalfa pellets (API-SARA). The population structure of the E. coli was evaluated with the ABD genotyping system and repetitive sequence-based (rep)-PCR fingerprinting. Twenty-five virulence factors were evaluated with PCR. Escherichia coli numbers were higher in the GPI-SARA treatment than in the API-SARA treatment. The genetic structure of the E. coli was significantly different between SARA challenge models. Isolates from GPI-control (46%), API-control (70%), and API-SARA (53%) were closely related and fell into one cluster, whereas isolates from GPI-SARA (54%) grouped separately. The ABD typing indicated a shift from an A-type E. coli population to a B1-type population only due to GPI-SARA. Of the 25 virulence factors tested, curli fiber genes were highly associated with GPI. Curli fibers were first identified in E. coli mastitis isolates and are potent virulence factors that induce a range of immune responses. Results suggest that under low rumen pH conditions induced by a grain diet, there is a burst in the number of E. coli with virulence genes that can take advantage of these rumen conditions to trigger an inflammatory response.

Morning vs. evening feed delivery for lactating dairy cows

Nikkhah, A., Furedi, C. J., Kennedy, A. D., Scott, S. L., Wittenberg, K. M. Crow, G. H. and Plaizier, J. C. 2011. Morning vs. evening feed delivery for lactating dairy cows. Can. J. Anim. Sci. 91: 113–122. A study was conducted to investigate the effects of changing feed delivery time from morning (0900) to evening (2100) on feed intake, diurnal feed intake patterns, digestion, rumen fermentation, milk production and composition, as well as on several blood metabolites. Eight (four ruminally cannulated) lactating Holstein cows, consisting of four primiparous and four multiparous cows, were used in the study. The change in the time of feed delivery increased the proportion of feed consumed within 3 h of feed delivery in all cows (54.7 vs. 46.3±4.1%, means±SE, P=0.05), but increased daily dry matter intake only in primiparous cows (18.5 vs. 20.4±0.7 kg d−1, P=0.05). Average daily concentrations of blood glucose, lactate, urea and β-hydroxybutyrate (BHBA) as well as rumen pH and concentrations of volatile fa...