Effects of California chaparral plants on in vitro ruminal fermentation of forage and concentrate diet

BACKGROUND: The combustible nature of chaparral plants has been attributed to the presence of secondary compounds such as phenolic acids, flavonoids and essential oils, among others. However, the implication of the antimicrobial properties of secondary compounds of chaparral in modulating rumen microbial metabolism has not been determined. The effects of 11 chaparral plants on rumen microbial fermentation were assessed in an in vitro batch culture fermentation fed a barley silage:barley grain‐based low concentrate (LC) and high concentrate (HC) diets. RESULTS: With LC, gas production [g dry matter (DM) incubated] and DM disappearance (DMD) were unaffected by Adenostoma fasciculatum, Ceanothus cuneatus, Baccharis pilularis or Eriodictyon californicum, but all plants except C. cuneatus decreased (P < 0.05) CH₄ production. With HC diet, all species except A. fasciculatum decreased (P < 0.01) total gas produced. Total volatile fatty acids (VFA) produced, molar proportions of acetate, propionate and butyrate, and the acetate:propionate (A:P) ratio were unaffected by chaparral addition to both HC and LC diets. Increased concentrations of A. fasciculatum and E. californicum decreased (linear; P < 0.05) fermentability and productions of CH₄ and NH₃‐N. CONCLUSION: Among all chaparral species investigated A. fasciculatum and E. californicum added as 10% of LC diet showed some potential to exert beneficial effects on rumen microbial fermentation. © Her Majesty the Queen in Right of Canada 2012     

Effect of essential oils on ruminal fermentation and lactation performance of dairy cows

Three experiments (Exp.) were conducted to study the effects of dietary addition of an essential oil product (EO) based on eugenol and cinnamaldehyde (0, control, or 525 mg/d of Xtract 6965; Pancosma SA, Geneva, Switzerland) on ruminal fermentation, total-tract digestibility, manure gas emissions, N losses, and dairy cow performance. In Exp. 1 and 3, the EO supplement was added to the vitamin-mineral premix. In Exp. 2, EO was top-dressed. Experiments 1 and 2 were crossover designs with 20 multiparous Holstein cows each (including 4 and 8 ruminally cannulated cows, respectively) and consisted of two 28-d periods. Intake of dry matter did not differ between treatments. Most ruminal fermentation parameters were unaffected by EO. Concentrations of ammonia (Exp. 1), isobutyrate (Exp. 1 and 2), and isovalerate (Exp. 1) were increased by EO compared with the control. Apparent total-tract digestibility of nutrients was similar between treatments, except total-tract digestibility of neutral-detergent fiber, which was increased or tended to be increased by EO in Exp. 1 and 2. Manure emissions of ammonia and methane were unaffected by EO. Blood plasma and milk urea-N concentrations and urinary N losses were increased by EO compared with the control in Exp. 1, but not in Exp. 2. Average milk yield, 3.5% fat-corrected milk yield, and milk fat, protein, and lactose concentrations were unaffected by treatment. Urinary excretion of purine derivatives, a marker for microbial protein production in the rumen, was greater in cows receiving the EO diet in Exp. 1, but not in Exp. 2. In Exp. 3, 120 Holstein cows were grouped in pens of 20 cows/pen in a 12-wk experiment to study production effects of EO. Dry matter intake, milk yield (a trend for a slight decrease with EO), milk components, milk urea N, and feed efficiency were similar between treatments. Results from these studies indicate that supplementing dairy cows with 525 mg/d of Xtract 6965 had moderate effects on ruminal fermentation, but consistently increased ruminal isobutyrate concentration and tended to increase total-tract digestibility of neutral-detergent fiber. Under the conditions of these experiments, Xtract 6965 fed at 525 mg/d did not affect milk production or composition.     

Effects of essential oils on rumen fermentation, milk production, and feeding behavior in lactating dairy cows

Seven ruminally cannulated lactating Holstein dairy cows were used in an incomplete Latin rectangle design to assess the effects of 2 commercial essential oil (EO) products on rumen fermentation, milk production, and feeding behavior. Cows were fed a total mixed ration with a 42:58 forage:concentrate ratio (DM basis). Treatments included addition of 0.5 g/d of CE Lo (85 mg of cinnamaldehyde and 140 mg of eugenol), 10 g/d of CE Hi (1,700 mg of cinnamaldehyde and 2,800 mg of eugenol), 0.25 g/d of CAP (50 mg of capsicum), or no oil (CON). Cows were fed ad libitum twice daily for 21 d per period. Dry matter intake, number of meals/d, h eating/d, mean meal length, rumination events/d, h ruminating/d, and mean rumination length were not affected by EO. However, length of the first meal after feeding decreased with addition of CE Hi (47.2 min) and CAP (49.4 min) compared with CON (65.4 min). Total volatile fatty acids, individual volatile fatty acids, acetate:propionate ratio, and ammonia concentration were not affected by EO. Mean rumen pH as well as bouts, total h, mean bout length, total area, and mean bout area under pH 5.6 did not differ among treatments. Total tract digestibility of organic matter, dry matter, neutral detergent fiber, acid detergent fiber, crude protein, and starch were not affected by EO. Milk yield and composition did not change with EO. In situ dry matter disappearance of ground soybean hulls was not affected by EO. However, organic matter disappearance of soybean hulls with CE Hi tended to decrease compared with CON. Compared with CON, neutral detergent fiber disappearance (41.5 vs. 37.6%) and acid detergent fiber disappearance (44.5 vs. 38.8%) decreased with addition of CE Hi. The CE Lo had no effect on rumen fermentation, milk production, or feeding behavior but CAP shortened the length of the first meal without changing rumen fermentation or production, making it a possible additive for altering feeding behavior. The CE Hi negatively affected rumen fermentation and shortened the length of the first meal, suggesting that a dose of 10 g/d is not beneficial to lactating dairy cows.     

Effect of essential oil active compounds on rumen microbial fermentation and nutrient flow in in vitro systems

Two experiments were conducted to determine the effects of several essential oil active compounds on rumen microbial fermentation. In the first experiment, 4 doses (5, 50, 500, and 5,000 mg/L) of 5 essential oil compounds were evaluated using in vitro 24-h batch culture of rumen fluid with a 60:40 forage:concentrate diet (18% crude protein; 30% neutral detergent fiber). Treatments were control (CON), eugenol (EUG), guaiacol, limonene, thymol (THY), and vanillin. After 24 h, the pH was determined, and samples were collected to analyze ammonia N and volatile fatty acids (VFA). The highest dose of all compounds decreased total VFA concentration and increased the final pH. Eugenol at 5 mg/L tended to reduce the proportion of acetate and the acetate to propionate ratio, at 50 and 500 mg/L tended to reduce ammonia N concentration, and at 500 mg/L reduced the proportion of propionate and branched-chain VFA concentration, without affecting total VFA concentration. All other treatments had minor effects or changes occurred only after total VFA concentration decreased. In the second experiment, 8 dual-flow continuous culture fermenters (1,320 mL) were used in 3 replicated periods (6 d of adaptation and 3 d of sampling) to study the effects of THY and EUG on rumen microbial fermentation. Fermenters were fed 95 g/d of DM of a 60:40 forage:concentrate diet (18% crude protein; 30% neutral detergent fiber). Treatments were CON, 10 mg/L of monensin (positive control), and 5, 50, or 500 mg/L of THY and EUG, and were randomly assigned to fermenters within periods. During the last 3 d of each period, samples were taken at 0, 2, 4, and 6 h after the morning feeding and analyzed for peptides, amino acids, and ammonia N concentrations, and total and individual VFA concentrations. Monensin changed the VFA profile as expected, but inhibited nutrient digestion. Eugenol and THY decreased total VFA concentration and changed the VFA profile, and only 5 mg/L of THY tended to reduce the proportion of acetate, increased the proportion of butyrate, and increased the large peptides N concentration without decreasing total VFA concentration. Most of these essential oil compounds demonstrated their antimicrobial activity by decreasing total VFA concentration at high doses. However, EUG in batch fermentation and 5 mg/L of THY in continuous culture modified the VFA profile without decreasing total VFA concentration, and EUG in batch fermentation decreased ammonia N concentration.     

Effects of replacing soybean meal with canola meal differing in rumen-undegradable protein content on ruminal fermentation and gas production kinetics using 2 in vitro systems

Previous research indicated that there were significant differences in rumen-undegradable protein (RUP) among canola meals (CM), which could influence the nutritional value of CM. The objectives of this study were to (1) evaluate the effects of feeding CM with different RUP contents on ruminal fermentation, nutrient digestion, and microbial growth using a dual-flow continuous culture system (experiment 1) and (2) evaluate ruminal gas production kinetics, in vitro organic matter (OM) digestibility, and methane (CH₄) production of soybean meal (SBM) and CM with low or high RUP in the diet or as a sole ingredient using a gas production system (experiments 2 and 3). In experiment 1, diets were randomly assigned to 6 fermentors in a replicated 3 × 3 Latin square. The only ingredient that differed among diets was the protein supplement. The treatments were (1) solvent-extracted SBM, (2) low-RUP solvent-extracted CM (38% RUP as a percentage of crude protein), and (3) high-RUP solvent-extracted CM (50% RUP). Diets were prepared as 3 concentrate mixtures that were combined with 25% orchardgrass hay and 15% wheat straw (dry matter basis). Experiments 2 and 3 had the same design with 24 bottles incubated 3 times for 48 h each. During the 48-h incubation, the cumulative pressure was recorded to determine gas production kinetics, in vitro OM digestibility, and CH₄ production. In experiment 1, N flow (g/d), efficiency of N use, efficiency of bacterial N synthesis, total volatile fatty acids (mM), and molar proportion of acetate, propionate, and isobutyrate were not affected by treatments. There were tendencies for a decrease in ruminal NH₃-N and an increase in molar proportion of butyrate for the SBM diet compared with both CM diets. The molar proportion of valerate was greater in both CM diets, whereas the molar proportion of isovalerate and total branched-chain volatile fatty acids was lower for the CM diets compared with the SBM diet. In experiments 2 and 3, the SBM diet had a greater gas pool size than both CM diets. The SBM diet increased in vitro OM digestibility; however, it also tended to increase CH₄ production (mM and g/kg of DM) compared with both CM diets. Based on the results of this study, CM with RUP varying from 38 to 50% of crude protein does not affect ruminal fermentation, nutrient digestion, and microbial growth when CM is included at up to 34% of the diet.     

Effects of cinnamon leaf, oregano and sweet orange essential oils on fermentation and aerobic stability of barley silage

BACKGROUND: Silage additives are marketed with the primary aim of improving the fermentation and/or aerobic stability of silage. The objective of this study was to evaluate the impact of three different essential oils (EOs; cinnamon leaf (CIN), oregano (ORE) and sweet orange (SO)) on the fermentation characteristics and stability of barley silage. Chopped whole‐plant barley (Hordem vulgare L.) forage was ensiled either untreated (0 mg kg^?1 dry matter (DM)) or treated with CIN, ORE or SO (37.5, 75 and 120 mg kg^?1 DM). RESULTS: Moulds were not detected in any treatments, including the control, after 7 days of air exposure. All EOs at a concentration of 120 mg kg^?1 silage DM decreased (P = 0.001) yeast populations in comparison with the control during air exposure. Net gas, methane and ammonia concentrations in vitro did not differ among treatments. Changes in volatile fatty acid concentrations were small, and in situ data showed no changes in DM and neutral detergent fibre digestion rates for CIN, ORE or SO at concentrations up to 120 mg kg^?1 DM. CONCLUSION: The findings from this study show that a concentration of 120 mg EO kg^?1 DM decreased yeast counts during aerobic stability tests. However, all EO treatments had minimal effects on data from in vitro and in situ incubations. Copyright © 2011 Crown in the right of Canada. Published by JohnWiley & Sons, Ltd     

Effects of addition of essential oils and monensin premix on digestion, ruminal fermentation, milk production, and milk composition in dairy cows

Four ruminally cannulated, lactating Holstein cows were used in a 4 × 4 Latin square design (28-d periods) with a 2 × 2 factorial arrangement of treatments to study the effects of dietary addition of essential oils (0 vs. 2 g/d; EO) and monensin (0 vs. 350 mg/d; MO) on digestion, ruminal fermentation characteristics, milk production, and milk composition. Intake of dry matter averaged 22.7 kg/d and was not significantly affected by dietary additives. Apparent digestibilities of dry matter, organic matter, neutral detergent fiber, and starch were similar among treatments. Apparent digestibility of acid detergent fiber was increased when diets were supplemented with EO (48.9 vs. 46.0%). Apparent digestibility of crude protein was higher for cows fed MO compared with those fed no MO (65.0 vs. 63.6%). Nitrogen retention was not changed by additive treatments and averaged 27.1 g/d across treatments. Ruminal pH was increased with the addition of EO (6.50 vs. 6.39). Ruminal ammonia nitrogen (NH₃-N) concentration was lower with MO-supplemented diets compared with diets without MO (12.7 vs. 14.3 mg/100 mL). No effect of EO and MO was observed on total volatile fatty acid concentrations and molar proportions of individual volatile fatty acids. Protozoa counts were not affected by EO and MO addition. Production of milk and 4% fat-corrected milk was similar among treatments (33.6 and 33.4 kg/d, respectively). Milk fat content was lower for cows fed MO than for cows fed diets without MO (3.8 vs. 4.1%). The reduced milk fat concentration in cows fed MO was associated with a higher level of trans-10 18:1, a potent inhibitor of milk fat synthesis. Milk urea nitrogen concentration was increased by MO supplementation, but this effect was not apparent when MO was fed in combination with EO (interaction EO × MO). Results from this study suggest that feeding EO (2 g/d) and MO (350 mg/d) to lactating dairy cows had limited effects on digestion, ruminal fermentation characteristics, milk production, and milk composition.     

Effect of humic acid on fermentation and ciliate protozoan population in rumen fluid of sheep in vitro

BACKGROUND: Humic acid (HA) as a product of decomposition of animal and plant tissue is used in animal production as a feed supplement, antimicrobial product and growth stimulator. The objective of the present in vitro study was to investigate the effects of dietary humic acid as a commercial Humacid 60 Basic (H60B) feed additive preparation on rumen fermentation and the ciliate protozoan population in the rumen fluid of sheep using a high fibre (HF) and high concentrate (HC) diet in batch cultures and artificial rumen (RUSITEC). RESULTS: Production of total volatile fatty acids (VFAs) was significantly decreased (P < 0.001) for batch cultures by the HF‐H60B diet. The HF‐H60B diet decreased ammonia N in RUSITEC (P < 0.001). An increase in the population of Enoploplastron triloricatum, Isotricha spp. and Ophryoscolex c. tricoronatus with the HF‐H60B diet and Diploplastron affine with the HC‐H60B was observed. The H60B did not affect the total ciliate population and Entodinium spp. CONCLUSION: It can be concluded that dietary humic acid preparations are not effective as dietary antiprotozoal agents. Humic acid might enhance microbial growth and energy efficiency in doses up to 10 g kg^?1 DM of diet. Copyright © 2009 Society of Chemical Industry     

Effects of pH and concentrations of linoleic and linolenic acids on extent and intermediates of ruminal biohydrogenation in vitro

Three experiments were conducted by in vitro incubations in ruminal fluid to investigate the effects of pH and amounts of linoleic and linolenic acids on the extent of their biohydrogenation, the proportions of conjugated linoleic acid (CLA) and trans-C18:1 as intermediates, and the ratio trans-10:trans-11 intermediates. The effects of pH and amount of linoleic acid were investigated in kinetic studies, and effects of the amount of linolenic acid were studied with 6-h incubations. With identical initial amounts of linoleic acid, its disappearance declined when the mean pH during incubation was under 6.0 compared with a mean pH over 6.5, and when the amount of linolenic acid increased from 10 to 180 mg/160-ml flask, suggesting an inhibition of the isomerization step of the biohydrogenation. Low pH decreased the ratio of trans-10:trans-11 intermediates. With initial amounts of linoleic acid increasing from 100 to 300 mg, the percentage of linoleic acid disappearance declined, but the amount that disappeared increased, without modification of the trans-10:trans-11 ratio, suggesting a maximal capacity of isomerization rather than an inhibition. Moreover, increasing initial linoleic acid resulted in high amounts of trans-C18:1 and an increase of C18:0 that was a linear function of time, suggesting a maximal capacity for the second reduction step of biohydrogenation. High amounts of initial linolenic acid did not affect the amounts of CLA, trans-C18:1, or the ratio trans-10:trans-11. Based on these experiments, a ruminal pH near neutrality with high amount of dietary linoleic acid should modulate the reactions of biohydrogenation in a way that supports CLA and trans-11C18:1 in the rumen.     

Supplementation of pearl millet flour with soybean protein: effect of cooking on in vitro protein digestibility and essential amino acids composition

In vitro protein digestibility (IVPD) and amino acids profile of pearl millet (Dempy cultivar) supplemented with soybean protein (5–15%) were investigated. Supplementation of dempy flour with soybean protein steadily decreased IVPD with increasing the portion of soybean in the blend. The in vitro protein digestibilities of the cooked supplemented dempy flours were higher when compared with the raw ones, whereas the highest value was that of the 5% soybean protein. All essential amino acids of dempy flour were enriched on supplementation with soybean protein. The levels of amino acids increased with increasing the amount of soybean protein in the blend. Essential amino acids in dempy supplemented with 15% soybean are comparable to those in the FAO reference pattern. Supplementation increased significantly lysine to 1.5–2.4 folds. Essential amino acids content remained higher in the cooked composite flours when compared with the cooked native dempy flour.     

Effect of sarsaponin on ruminal fermentation with particular reference to methane production in vitro

This experiment was designed to investigate the effects of different concentrations (0, 1.2, 1.8, 2.4, and 3.2 g/L) of sarsaponin on ruminal microbial methane production using the substrates soluble potato starch, cornstarch, or hay plus concentrate (1.5:1). Ruminal fluid was collected from a dairy cow, mixed with phosphate buffer (1:2) and incubated (30 ml) anaerobically at 38 degrees C for 6 and 24 h with or without sarsaponin. Excluding the lower level of sarsaponin, pH of the medium was slightly decreased. Ammonia-N concentration and numbers of protozoa were decreased in a dose-dependent manner. Total volatile fatty acids and total gas production were increased. Molar proportion of acetate was decreased and propionate was increased with a corresponding decrease in acetate:propionate ratio. Hydrogen production was decreased. As the concentration of sarsaponin increased from 1.2 to 3.2 g/L, fermentation of soluble potato starch, cornstarch, or hay plus concentrate decreased methane production from 20 to 60% (6 h) and 17 to 50% (24 h), 21 to 58% (6 h) and 18 to 52% (24 h), and 23 to 53% (6 h) and 15 to 44% (24 h), respectively. Excluding the lower dose concentration (1.2 g/L) of sarsaponin, in vitro disappearance of dry matter of hay plus concentrate was decreased after 24 h. In conclusion, these results show that sarsaponin stimulated the mixed ruminal microorganism fermentation as well as to inhibit methane production in vitro.     

Milk odd- and branched-chain fatty acids in relation to the rumen fermentation pattern

The objectives of this study were 1) to determine whether a relationship exists between molar proportions of volatile fatty acids in the rumen and milk odd-and branched-chain fatty acid concentrations (i.e., iso C13:0, anteiso C13:0, iso C14:0, C15:0, iso C15:0, anteiso C15:0, iso C16:0, C17:0, iso C17:0, anteiso C17:0, and cis-9 C17:1); and 2) to evaluate the accuracy of prediction of the latter equations using an independent data set. For development of the regression equations, individual cow data from 10 feeding experiments with rumen-fistulated dairy cows were used, resulting in a data set of 148 observations. Milk odd- and branched-chain fatty acids were closely related to the molar proportions of acetate (SE = 15.3 mmol/mol), propionate (SE = 14.7 mmol/mol), and butyrate (SE = 9.2 mmol/mol). These regression equations were further validated using data from the literature (n = 14). Evaluation of these prediction equations using the independent data set resulted in a root mean square prediction error of 3.0, 9.0, and 8.9% of the observed mean for acetate, propionate, and butyrate, respectively. In addition, less then 5% of the mean square prediction error was due to line bias. This suggests that the currently developed prediction equations based on milk odd- and branched-chain fatty acids show potential to predict molar proportions of individual volatile fatty acids in the rumen.     

Sources of variation in rates of in vitro ruminal protein degradation

Rates and extents of ruminal protein degradation for casein, solvent soybean meal (SSBM), expeller soybean meal (ESBM), and alfalfa hay were estimated from net appearance of NH3 and total amino acids in in vitro media containing 1 mM hydrazine and 30 mg/L of chloramphenicol. Protein was added at 0.13 mg of N/mL of medium, and incubations were conducted for 4 to 6 h, usually with hourly sampling. Inocula were obtained from ruminally cannulated donor cows fed diets of grass silage or alfalfa and corn silages plus concentrates. Preincubation or dialysis of inocula was used to suppress background NH3 and total amino acids; however, preincubation yielded more rapid degradation rates for casein and SSBM and was used in subsequent incubations. Preincubation with added vitamins, VFA, hemin, or N did not alter protein degradation. Protein degradation rates estimated for SSBM, ESBM, and alfalfa were not different when computed from total N release or N release in NH3 plus total amino acids, regardless of whether amino acids were quantified using ninhydrin colorimetry or o-phthalaldehyde fluorescence. Accounting for the release of peptide-N also did not affect estimated degradation. However, casein degradation rates were more rapid when using total N release or accounting for peptide-N, indicating significant accumulation of small peptides during its breakdown. Rates also were more rapid with inocula from lactating cows versus nonlactating cows with lower feed intake. Protein degradation rates were different due to time after feeding: casein rate was more rapid, but SSBM and ESBM rates were slower with inocula obtained after feeding. Several characteristics of ruminal inoculum that influenced breakdown of the rapidly degraded protein casein did not appear to have direct effects on degradation of protein in soybean meal.     

Effects of essential oils on milk production, milk composition, and rumen microbiota in Chios dairy ewes

The effect of the addition of an essential oil (EO) preparation (containing a mixture of natural and nature-identical EO) on the performance of dairy ewes of the Chios breed was investigated. Eighty lactating ewes were allocated into 4 equal groups in a randomized block design, each with 4 replicates of 5 ewes housed in the same pen. The 4 groups were fed the same total mixed ration allowance, the roughage being a mixture of corn silage, lucerne hay, and wheat straw, and the concentrate based on cereals and oil cakes. Control ewes were fed their daily allowance of total mixed ration without any EO. The other 3 groups were supplemented with EO at levels of 50, 100, and 150 mg/kg of the concentrated feed, respectively. Individual milk yield was recorded daily and feed refusals were recorded on a pen basis weekly during the first 5 mo of lactation. Milk samples were analyzed for chemical composition, somatic cell count, and urea content. Rumen samples were analyzed for pH, NH₃-N content, and protozoa, cellulolytic, hyper-ammonia-producing, and total viable bacteria counts. Results showed that inclusion of EO increased milk production per ewe, the effect being dose dependent [1.565, 1.681, 1.876, and 2.119 L/d (standard error of the difference ± 0.176) for the control, 50, 100, and 150 mg of EO/kg of concentrate diets, respectively], and thus improved feed utilization. Although the inclusion of EO did not affect milk composition, it lowered urea concentration and somatic cell count in milk samples at the highest supplementation level compared with the control. Total counts of viable and cellulolytic bacteria and protozoa were not influenced by EO supplementation; however, counts of hyper-ammonia-producing bacteria were decreased at the 2 highest supplementation levels compared with the control group. Rumen pH was not affected by EO supplementation, but rumen NH₃-N was reduced at the highest EO supplementation level, and acetate rumen concentrations tended to decrease and propionate to increase in a dose-dependent manner. In conclusion, EO supplementation may improve feed utilization and performance of the high-yielding dairy Chios ewes; however, the underlying mechanisms leading to this improvement merit further investigation.     

Effects of partial replacement of corn grain with lactose in calf starters on ruminal fermentation and growth performance

The objective of this study was to evaluate effects of partial replacement of dry ground corn with lactose in calf starters on dry matter intake, growth rate, ruminal pH, and volatile fatty acid profile. Sixty Holstein bull calves were raised on a high plane of nutrition program until 55 d of age. Calves were fed texturized calf starters containing 30.1% steam-flaked grains and lactose at 0 (control), 5, or 10% (n = 20 for each treatment) on a dry matter basis. All calves were fed treatment calf starters ad libitum from d 7 and kleingrass hay from d 35. Ruminal pH was measured continuously immediately after weaning (d 55–62) for 15 calves (n = 5 per treatment), and 3 wk after weaning (d 77 to 80) for the other 45 calves (n = 15 per treatment). Dry matter intake, growth performance, and ruminal pH variables were not affected by treatment. However, according to Spearman's correlation coefficient (r_s) analyses, lactose intake was positively correlated with dairy minimum ruminal pH (r_s = 0.306) for the data collected from d 77 to 80. Similarly, hay intake was not affected by treatment, but positively correlated with daily mean (r_s = 0.338) and maximum ruminal pH (r_s = 0.408) and negatively correlated with duration pH <5.8 (r_s = ?0.329) and area pH <5.8 (r_s = ?0.325), indicating that the variation in hay intake among animals might have masked treatment effects on ruminal pH. Ruminal molar ratio of acetate was higher (45.2 vs. 40.6%), and that of propionate was lower in 10% lactose than control (35.3 vs. 40.2%) for ruminal fluid collected on d 80; however, molar ratio of butyrate was not affected by treatment. These results indicate that lactose inclusion in calf starters up to 10% of dry matter might not affect dry matter intake and growth performance of calves, but that greater lactose and hay intake might be associated with higher ruminal pH.