Enzyme supplementation to improve the nutritional value of fibrous feed ingredients in swine diets fed in dry or liquid form.

This study evaluated the effect of xylanase supplementation (with or without), feeding method (dry or liquid), and feedstuff (corn distiller's dried grains with solubles [DDGS] or wheat middlings) on apparent total tract digestibility (ATTD) and apparent ileal digestibility (AID) of GE and nutrients, intestinal morphology, ileal and cecal pH, and VFA concentrations. Sixty-four growing pigs (25.87 ± 0.38kg initial BW) were blocked by BW and sex and randomly assigned to 8 dietary treatments. Within each feedstuff, diets were fed either liquid or dry, without or with xylanase (24,000 birch xylan units/kg feed), for 16 d. Diets contained 3.32 and 3.19 Mcal/kg ME for DDGS- and wheat middlings-based diets, respectively. Pigs were fed restricted at 3 times maintenance ME requirements. Liquid diets were prepared by steeping DDGS or wheat middlings with water (1:3, wt/vol) with or without xylanase for 24 h followed by mixing with a basal ingredient mixture and water to achieve a final ratio of 1:2.5 (wt/vol). During steeping of wheat middlings, some fiber degradation occurred. When xylanase was added in dry wheat middlings diets, AID of GE ( < 0.10) and NDF ( < 0.05) increased compared with dry wheat middlings diets without xylanase (64.50 vs. 54.67% and 52.88 vs. 31.69%, respectively), but supplementation of xylanase did not impact AID of GE and NDF when liquid wheat middlings diets were fed. Xylanase in liquid DDGS diets increased ( < 0.05) the AID of NDF compared with liquid DDGS diets without xylanase, but xylanase did not affect AID of NDF in dry DDGS diets. Xylanase in wheat middlings diets improved ( < 0.05) ATTD of GE and N compared with wheat middlings diets without xylanase (80.37 vs. 78.07% and 80.23 vs. 77.94%, respectively). However, there was no effect of xylanase in DDGS diets. Pigs fed DDGS diets had greater concentrations of butyrate in the cecum ( = 0.001) than pigs fed wheat middlings diets (27.6 vs. 20.4 mmol/L). Pigs fed DDGS diets with xylanase had deeper crypts ( < 0.05) in the jejunum than pigs fed DDGS diets without xylanase (98.20 vs. 86.16 µm), but xylanase had no effect in pigs fed wheat middlings diets. Results suggest that liquid feeding and xylanase supplementation had limited potential to enhance nutrient digestibility in pigs fed DDGS-based diets. However, xylanase supplementation in dry wheat middlings-based diets improved the AID of NDF and ATTD of GE and N, but liquid feeding as pretreatment did not further enhance the nutritional value of wheat middlings-based diets.

Impact of dietary lipids on sow milk composition and balance of essential fatty acids during lactation in prolific sows.

Two studies were designed to determine the effects of supplementing diets with lipid sources of EFA (linoleic and α-linolenic acid) on sow milk composition to estimate the balance of EFA for sows nursing large litters. In Exp. 1, 30 sows, equally balanced by parity (1 and 3 to 5) and nursing 12 pigs, were fed diets supplemented with 6% animal-vegetable blend (A-V), 6% choice white grease (CWG), or a control diet without added lipid. Diets were corn-soybean meal based with 8% corn distiller dried grains with solubles and 6% wheat middlings and contained 3.25 g standardized ileal digestible Lys/Mcal ME. Sows fed lipid-supplemented diets secreted greater amounts of fat (P = 0.082; 499 and 559 g/d for control and lipid-added diets, respectively) than sows fed the control diet. The balance of EFA was computed as apparent ileal digestible intake of EFA minus the outflow of EFA in milk. For sows fed the control diet, the amount of linoleic acid secreted in milk was greater than the amount consumed, throughout lactation. This resulted in a pronounced negative balance of linoleic acid (-22.4, -38.0, and -14.1 g/d for d 3, 10, and 17 of lactation, respectively). In Exp. 2, 50 sows, equally balanced by parity and nursing 12 pigs, were randomly assigned to a 2 × 2 factorial arrangement of diets plus a control diet without added lipids. Factors included linoleic acid (2.1% and 3.3%) and α-linolenic acid (0.15% and 0.45%). The different concentrations of EFA were obtained by adding 4% of different mixtures of canola, corn, and flaxseed oils to diets. The n-6 to n-3 fatty acid ratios in the diets ranged from 5 to 22. Increasing supplemental EFA increased (P < 0.001) milk concentrations of linoleic (16.7% and 20.8%, for 2.1% and 3.3% linoleic acid, respectively) and α-linolenic acid (P < 0.001; 1.1 and 1.9% for 0.15 and 0.45% α-linolenic acid, respectively). Increasing supplemental EFA increased the estimated balance of α-linolenic acid (P < 0.001; -0.2 and 5.3 g/d for 0.15% and 0.45% α-linolenic acid, respectively), but not linoleic acid (P = 0.14; -3.4 and 10.0 g/d for 2.1% and 3.3% linoleic acid, respectively). In conclusion, lipid supplementation to sow lactation diets improved milk fat secretion. The fatty acid composition of milk fat reflected the dietary supplementation of EFA. The net effect of supplemental EFA was to create a positive balance during lactation, which may prove to be beneficial for the development of nursing piglets and the subsequent reproduction of sows.

Eicosapentaenoic and docosahexaenoic acids increase insulin sensitivity in growing steers.

An experiment was conducted to determine the effect of dietary n-3 long chain PUFA on insulin sensitivity in growing steers. Steers (n = 12, initial BW = 336.3 kg, SEM = 7.7) were adapted to a basal diet that was 70% concentrate mix and 30% orchardgrass hay. Steers were fed a daily amount of 0.26 Mcal ME per kg BW (0.75). After 3 wk steers were transitioned to 1 of 2 treatment (Trt) diets (n = 6 per diet) containing added Ca salts of fatty acids at 4% of DM using a source of fat that was enriched in n-3 fatty acids, including eicosapentaenoic acid and docosahexaenoic acid (FOFA), or a source of fat without n-3 fatty acids and a greater percentage of C16:0 and C18:1 (LCFA). Three intravenous (i.v.) glucose tolerance tests (IVGTT) were conducted, 1 during the basal diet, and 2 after transition to treatment diets at time 1 (T1; d 4 Trt) and time 2 (T2; d 39 Trt). Three i.v. insulin challenge tests (IC) were conducted the day after each IVGTT. Measurements on the basal diet were used as covariates. For IVGTT, there was a diet by time interaction (P < 0.05) for glucose area under the response curve (AUC). The AUC50 (mM glucose × 50 min) at T1 was less (P = 0.02) for LCFA (126.2) than FOFA (151.8), AUC50 at T2 tended to be greater (P = 0.07) for LCFA (165.9) than FOFA (146.0). Preinfusion insulin concentration was greater (P < 0.001) before the IVGTT and IC for steers fed LCFA (40.4 and 40.2 µIU/mL) than for steers fed FOFA (23.7 and 27.1 µIU/mL), respectively. Glucose clearance did not differ between treatments. For IC, minimum glucose concentration was greater (P = 0.02) and glucose AUC150 was less (P < 0.01) for steers fed LCFA than for steers fed FOFA. Values for glucose concentration were 1.8 mM and 1.5 mM and for AUC150 (mM glucose × 150 min) were 203.1 and 263.6 for steers fed LCFA and FOFA, respectively. Insulin clearance (fraction/min) was greater (P < 0.01) for steers fed LCFA (0.121) than FOFA (0.101). The insulin AUC60 (µIU/mL × 60 min) postinfusion was less for LCFA (17,674) than FOFA (19,997), and tended to be greater (P = 0.06) at T1 (19,552) than T2 (18,119). Plasma NEFA concentrations at T2 during IVGTT were greater (P < 0.05) in steers fed FOFA than in steers fed LCFA. The results indicated that supplementing with long-chain n-3 PUFA (FOFA) increased insulin sensitivity in growing steers compared with feeding a supplement with greater concentration of SFA (LCFA).

Ruminal fermentation, milk fatty acid profiles, and productive performance of Holstein dairy cows fed 2 different safflower seeds.

A lactation trial was conducted to determine the effects of supplementing whole safflower seeds (SS) on ruminal fermentation, lactational performance, and milk fatty acid (FA) profiles. Nine multiparous Holstein cows (days in milk = 110 ± 20) were used in a replicated 3 × 3 Latin square design. Each period lasted 21 d, with 14 d of adaptation and 7 d of data collection. Within square, cows were randomly assigned to a sequence of 3 dietary treatments as follows: cottonseed total mixed ration (TMR; CST), conventional SS (variety S-208) TMR (CSST), and NutraSaff SS (Safflower Technologies International, Sidney, MT) TMR (NSST). Diets contained approximately 63% forage (36% alfalfa hay, 4% grass hay, and 23% corn silage) and 37% concentrate supplemented with 2% cottonseed to the CST and 3% conventional or NutraSaff SS to the CSST or the NSST, respectively. Intake of dry matter (DM) averaged 21.8 kg/d and did not differ across diets, but feeding the NSST decreased intake of neutral detergent fiber (NDF) due to lower dietary concentration of NDF in the NSST. Digestibilities of DM and nutrients were similar among treatments. No differences in yields of milk or milk components were observed in response to supplementing SS. Dietary treatments did not affect ruminal pH, total or molar proportions of ruminal volatile FA, and ammonia-N. However, cows fed SS had a higher molar proportion of isobutyrate than those fed the CST diet. Ruminal C16:0 FA concentration increased with the CST, whereas C18:1 cis-9 and C18:2 n-6 tended to increase with SS supplementation, indicating that conventional and NutraSaff SS were partially protected from microbial biohydrogenation. Supplementing SS decreased milk C16:0 concentration, whereas it increased C18:1 cis-9 and C18:1 trans-9. Milk FA C18:1 trans-11 and cis-9, trans-11 conjugated linoleic acid increased and tended to increase with feeding the NSST, respectively, but not the CSST diet. In conclusion, supplementing diets with whole SS at 3% of dietary DM can be an effective strategy of fat supplementation to lactating dairy cows without negative effects on lactational performance and milk FA profiles.

Effects of supplementing dairy cows with chromium propionate on milk and tissue chromium concentrations.

Eight primiparous and 8 multiparous Holstein cows were used to determine the effects of Cr supplementation, in the form of Cr propionate (Cr Prop), on milk and tissue Cr concentrations. Cows were randomly assigned by parity to one of 2 diets: 1) control diet or 2) 2 mg of supplemental Cr/kg of DM. The level of Cr Prop supplemented exceeded by 4-fold the concentration of 0.5 mg of Cr/kg permitted by the FDA. Experimental diets were fed from approximately 30 d prepartum until at least 91 d postpartum, resulting in a minimum of 121 d of exposure to supplemental Cr. The control prepartum and postpartum diets analyzed 0.48 and 0.38 mg of Cr/kg of DM, respectively. Milk samples were obtained from the a.m. milking on d 0 (colostrum), 7, 14, 21, 28, 42, 56, 77, and 90 and on the final day of the study for Cr analysis. Cows were harvested after lactating for a minimum of 91 d and samples of liver, kidney, semitendinosus muscle, and fat were obtained for Cr analysis. Chromium was measured using electrothermal atomic absorption spectrophotometry. Milk Cr concentration averaged 1.7 ng/mL and was affected by day of lactation but not by Cr or a Cr × day interaction. Supplementation of 2 mg of Cr/kg of DM increased kidney Cr by approximately 3-fold and liver Cr concentrations by approximately 2-fold. Chromium concentrations in muscle and fat were not affected by Cr supplementation. In summary, supplementation of Cr Prop at a level of 2 mg of Cr/kg of DM did not affect Cr concentration in milk, muscle, or fat, the major bovine products consumed by humans.

Effects of antibiotics and oil on microbial profiles and fermentation in mixed cultures of ruminal microorganisms.

Ionophores and supplemental fat are fed to lactating cows to improve feed efficiency. Their effect on rumen fermentation is similar, but less is known about their impact on rumen microbes. The objective of this study was to determine the effects of monensin (M), bacitracin (B), and soybean oil (O) on microbial populations. Mixed cultures of rumen microbes were incubated in 5 dual-flow continuous fermentors and fed 13.8 g of alfalfa hay pellets daily (DM basis) for 16 d. All fermentors were allowed to stabilize for 4 d. From d 5 to 10, two fermentors received O (5% of diet DM), one fermentor received M (22 mg/kg), and one received B (22 mg/kg). From d 11 to 16, the 2 fermentors receiving O also received either M (OM) or B (OB) and O was included in the fermentors receiving M (MO) and B (BO). One fermentor served as the control and received 100% alfalfa pellets throughout the experiment. Each run was replicated 3 times. Samples were taken at 2 h after the morning feeding on d 4, 10, and 16 and were analyzed for bacterial populations using terminal restriction fragment length polymorphism. Volatile fatty acid concentration, methane production, and pH in the control cultures were not affected by time and remained similar during the entire experiment. The M and O treatments reduced molar concentration of acetate, increased concentration of propionate, and decreased methane production. Bacitracin did not alter acetate or propionate concentration, but reduced methane production. All 3 treatments (M, B, and O) altered the fragment patterns of microbial profiles. In contrast, treatments MO, OM, BO, and OB had little effect on culture fermentation despite differences in the patterns of microbial fragments. The terminal restriction fragment length polymorphism data suggest that microbial adaptation to the in vitro system in the control fermentor occurred within 4 d.

Supplementing limited methionine diets with rumen-protected methionine, betaine, and choline in early lactation Holstein cows.

Eighty lactating Holstein cows from 21 to 91 d in milk were fed a corn silage-based total mixed ration (TMR) formulated with the Met content limited (42 g/ d) to investigate the impact of supplementing rumen-protected (RP) forms of Met, betaine, and choline on performance and metabolism. One of 4 supplements was blended into the TMR to produce 4 dietary treatments: 1) control, 2) 20 g/d of RP-Met, 3) 45 g/d of RP-betaine, and 4) 40 g/d of RP-choline. Calcium salts of fatty acids were used to protect both RP-betaine and RP-choline supplements. A similar amount of Ca salts of fatty acids was included in both control and RP-Met supplements to provide equal amounts of fat to all treatments. Overall, no differences in intake, milk yield, or milk composition were observed in primiparous cows. Average dry matter intake, body weight, and body condition score were not different among treatments in multiparous cows. Milk yield was higher in multiparous cows fed RP-choline compared with the other treatments. Multiparous cows fed RP-choline had higher milk protein yield than cows fed control or RP-betaine but was not different from cows fed RP-Met. Multiparous cows fed RP-choline had higher milk fat yield than cows fed RP-Met but was not different from cows fed control or RP-betaine. There were no beneficial effects of RP-betaine supplementation to a Met-limited TMR.

Influence of cobalt concentration on vitamin B12 production and fermentation of mixed ruminal microorganisms grown in continuous culture flow-through fermentors.

An experiment was conducted to determine the effects of dietary concentrations of Co on vitamin B12 production and fermentation of mixed ruminal microbes grown in continuous culture fermentors. Four fermentors were fed 14 g of DM/d. The DM consisted of a corn and cottonseed hull-based diet with Co supplemented as CoCO3. Dietary treatments were 1) control (containing 0.05 mg of Co/kg of DM), 2) 0.05 mg of supplemental Co/kg of DM, 3) 0.10 mg of supplemental Co/kg of DM, and 4) 1.0 mg of supplemental Co/kg of DM. After a 3-d adjustment period, fermentors were sampled over a 3-d sampling period. This process was repeated 2 additional times for a total of 3 runs. Ruminal fluid vitamin B12 concentrations were affected by Co supplementation (P < 0.01), and there was a treatment x day interaction (P < 0.01). By sampling d 3, cultures fed the basal diet supplemented with 0.10 mg of Co/kg had greater (P < 0.05) vitamin B12 concentrations than those supplemented with 0.05 mg of Co/kg of DM, and increasing supplemental Co from 0.10 to 1.0 mg/kg of DM increased (P < 0.01) ruminal fluid vitamin B12 concentration. Ruminal fluid succinate also was affected (P < 0.10) by a treatment x day interaction. Cobalt supplementation to the control diet greatly decreased (P < 0.05) succinate in ruminal cultures on sampling d 3 but not on d 1 or 2. Molar proportions of acetate, propionate, and isobutyrate, and acetate:propionate were not affected by the addition of supplemental Co to the basal diet. However, molar proportions of butyrate, valerate, and isovalerate increased (P < 0.05) in response to supplemental Co. The majority of long-chain fatty acids observed in this study were not affected by Co supplementation. However, percentages of C18:0 fatty acids in ruminal cultures tended (P < 0.10) to be greater for Co-supplemented diets relative to the control. Methane, ammonia, and pH were not greatly affected by Co supplementation. The results indicate that a total (diet plus supplemental) Co concentration of 0.10 to 0.15 mg/kg of dietary DM resulted in adequate vitamin B12 production to meet the requirements of ruminal microorganisms fed a high-concentrate diet in continuous-flow fermentors.

Monensin by fat interactions on trans fatty acids in cultures of mixed ruminal microorganisms grown in continuous fermentors fed corn or barley.

In previous studies, monensin (M) and unsaturated plant oils independently increased trans fatty acid concentrations in cultures of mixed ruminal microorganisms. This study was conducted to determine if combining M with plant oil yielded interactions on trans fatty acid concentrations in cultures of mixed ruminal microorganisms or their effects were additive. Four continuous fermentors were fed 14 g of dry feed per day (divided equally between two feedings), consisting of alfalfa hay pellets (30% of DM) and either a high corn (HC) or a high barley (BB) concentrate (70% of DM) in each of two fermentors. Within each grain type, one fermentor was supplemented with M (25 ppm), and the other fermentor was supplemented with 5% soybean oil (SBO) during d 5 to 8. Monensin and SBO were added together in all fermentors during d 9 to 12. Samples were taken at 2 h after the morning feeding on the last day of each period and analyzed for fatty acids by gas-liquid chromatography. A second run of the fermentors followed the same treatment sequence to give additional replication. Average pH across all treatments was 6.15, which was reduced by M but not affected by SBO. Monensin reduced the ratio of acetate to propionate (A:P), which averaged 2.03 across all treatments; fat decreased A:P in cultures not receiving M but increased it in the presence of M. Monensin and SBO altered the concentration of several trans fatty acids, but the only interaction was a grain x M x SBO interaction for trans-10 C18:1. The increase in trans-10 C18:1 by the M and SBO combination exceeded the sum of increases in trans-10 C18:1 for each individual feed additive, but only for KB. For the HC diet, M increased trans-10 C18:1 more than fat alone and more than the M and SBO combination. The results of this study show that M and SBO effects are additive for all trans FA except for trans-10 C18:1. In the case of trans-10 C18:1, M and SBO interacted to give higher trans-10 C18:1 concentrations in ruminal contents than would be expected simply by adding their individual effects, but only for HB. Because some trans fatty acid isomers have been associated with milk fat depression in dairy cows, these results suggest more severe depressions in milk fat content when cows are fed M along with unsaturated plant oils.

Copper status, serum cholesterol, and milk fatty acid profile in Holstein cows fed varying concentrations of copper.

An experiment was conducted to determine the effects of dietary copper (Cu) on Cu status and lipid metabolism in Holstein cows. Three primiparous and 21 multiparous Holstein cows were utilized in this experiment. Groups of three cows similar in parity, days in milk, and milk yield were assigned randomly to one of the following three treatments: 1) control (no supplemental Cu), 2) 10 mg of Cu/kg of DM from Cu sulfate (CuSO4), and 3) 40 mg of Cu/kg of DM from CuSO4. Liver Cu concentrations were higher in Cu supplemented cows at the end of the 61-d study. Cows receiving 40 mg of Cu/kg of DM had higher liver Cu concentrations than cows receiving 10 mg of Cu. Plasma Cu concentrations were similar across treatments. Total serum cholesterol concentrations were higher in cows receiving supplemental Cu. Cows receiving 40 mg of Cu/kg of DM had higher serum cholesterol concentrations than cows receiving 10 mg of Cu. Dry matter intake, average daily milk production, and milk lipid, protein, and somatic cell numbers were similar across treatments. On d 61, milk fatty acids C18:1 trans and C18-conjugated dienes were lower in cows receiving supplemental Cu relative to the nonsupplemented controls. Cows receiving 40 mg of Cu/kg of DM had higher C12:0 and lower C18:2 and total polyunsaturated fatty acids in milk than cows receiving 10 mg of Cu/kg of DM. These results indicate that Cu supplementation alters lipid metabolism in high producing dairy cows and that Cu supplementation at 40 mg/kg of DM for 61 d can elevate liver Cu concentrations to levels considered to be marginally toxic in dairy cattle.

Effects of soybean oil and dietary copper on ruminal and tissue lipid metabolism in finishing steers.

An experiment was conducted to determine the effects of Cu and soybean oil (SBO) supplementation on ruminal and tissue lipid metabolism and carcass characteristics in finishing steers. Sixty Angus steers (369.0 +/- 10.1 kg) were stratified by weight and randomly assigned to treatments in a 2 x 2 factorial arrangement, with factors being 0 or 20 mg of supplemental Cu/kg DM from Cu sulfate and 0 or 4% SBO. Steers were fed a high-concentrate basal diet that contained 5.3 mg Cu/kg DM. Average daily gain and feed intake were reduced (P < 0.01) by SBO but were not affected by Cu. Gain:feed ratio was not affected by treatment. Liver Cu concentrations were higher (P < 0.01) in steers receiving supplemental Cu and lower (P < 0.04) in SBO-supplemented steers. Copper supplementation tended to reduce (P < 0.12) and SBO supplementation tended to increase (P < 0.11) serum cholesterol concentrations. Backfat depth was reduced (P < 0.10) by Cu and SBO supplementation. Marbling scores and longissimus muscle lipid content were not affected by Cu supplementation; however, SBO supplementation reduced (P < 0.01) marbling scores. Longissimus muscle polyunsaturated fatty acids tended to be increased (P < 0.14) in Cu-supplemented steers. Longissimus muscle C18-conjugated dienes and the 18:1 trans isomer were increased (P < 0.05) in SBO-supplemented steers. Ruminal fluid 18:3 was increased (P < 0.05) and the 18:1 trans isomer was decreased (P < 0.05) in Cu-supplemented steers. These results indicate that as little as 20 mg of supplemental Cu/kg DM can reduce backfat and may alter lipid metabolism in steers fed high-concentrate diets.

Methane output and lactation response in Holstein cattle with monensin or unsaturated fat added to the diet.

We measured effects of continuous vs twice-daily feeding, the addition of unsaturated fat to the diet, and monensin on milk production, milk composition, feed intake, and CO2-methane production in four experiments in a herd of 88 to 109 milking Holsteins. Methane and CO2 production increased with twice-daily feeding, but the CO2:CH4 ratio remained unchanged. Soybean oil did not affect the milkfat percentages, but fatty acid composition was changed. All saturated fatty acids up to and including 16:0 decreased (P < .01), whereas 18:0 and trans 18:1 increased (P < .001). The 18:2 conjugated dienes also increased (P < .01) when the cows were fed soybean oil. Monensin addition to the diet at 24 ppm decreased methane production (P < .01); the CO2:CH4 ratios reached 15, milk production increased (P < .01), and milkfat percentage and total milkfat output decreased (P < .01), as did feed consumption, compared with cows fed diets without monensin (P < .05). Milk fatty acid composition showed evidence of depressed ruminal biohydrogenation: saturated fatty acids (P < .05) decreased and 18:1 increased (P < .001); most of the increase was seen in the trans 18:1 isomer. As with soybean oil feeding, addition of monensin also increased (P < .05) the concentration of conjugated dienes. The monensin feeding trial was repeated 161 d later with 88 cows, of which 67 received monensin in the diet in the first trial and 21 cows were newly freshened and had never received monensin. Methane production again decreased (P < .05), but this time the CO2:CH4 ratio did not change and all other monensin-related effects were absent. The ruminal microflora in the cows that had previously received monensin seemed to have undergone some adaptive changes and no longer responded as before.

Comparative assessment of bacterial inoculation and propionic acid treatment of aerobic stability and microbial populations of ensiled high-moisture ear corn.

High-moisture ear corn (HMEC) was untreated, treated with propionic acid (PA), or inoculated with a mixture of Lactobacillus plantarum and Enterococcus faecium and allowed to ensile in laboratory silos for 0, 7, 21, 42, 138, or 202 d. The silages were evaluated for fermentation quality, microbial populations, and aerobic stability. In all treatments, silage pH declined rapidly within 7 d, but the rate of decline seemed greatest with the inoculum. The lactic acid content of inoculated HMEC was higher (P < .05) than that of control of PA-treated HMEC. Regardless of treatment, the population of lactic acid bacteria (LAB) increased (P < .1) up to 7 to 21 d of fermentation then declined; LAB counts decreased (P < .05) up to 42 d in control and PA-treated silage but continued to decline until 138 d for inoculated silage. Yeast and mold counts tended to decrease up to 42 d of ensiling then decreased (P < .05) as fermentation progressed. Between 138 and 202 d of ensiling, the control silage showed a marked increase (P < .10) in pH and yeast and mold populations, providing evidence of secondary fermentation; PA treatment and bacterial inoculation prevented secondary fermentation. Inoculation tended to reduce estimates of sample temperature for silage stored for 138 d and exposed to air, but not for the corresponding silage stored for 202 d. Treatment with PA prevented the loss (P > .05) of acetic acid and the rise (P > .05) in pH during air exposure of the 138-d silage; both control and PA-treated silage showed an increase (P < .05) in yeast and mold populations, but the increments were 38% and 23%, respectively. Compared with PA, the relative efficacy of inoculation in improving aerobic spoilage of HMEC depended on the period of silo storage and the criterion used to assess aerobic stability.