Silage review: Interpretation of chemical, microbial, and organoleptic components of silages.

The goal of making silage is to produce a stable feed with a high recovery of dry matter, energy, and highly digestible nutrients compared with the fresh crop. Microbial fermentation in the silo produces an array of end products and can change many nutritive aspects of a forage. High-quality silage should be void of undesirable compounds that could negatively affect animal performance, the environment, or net farm income. This review discusses the interpretation of the common fermentation end products, microbial populations, organoleptic properties, and changes in nutritive aspects of silages during storage of silages with emphasis on a North American perspective.

Silage review: Factors affecting dry matter and quality losses in silages.

An overview was made of dry matter (DM) and quality losses that occur during the ensiling process from the field through the feeding phase. The aim was to review the relevant published literature of the last 15 yr focusing on developments achieved after the publication of the book Silage Science and Technology. This review discusses the factors affecting DM and quality losses in terms of field and pre-ensiling conditions, respiration and temperature at ensiling, fermentation patterns, methods of covering and weighting the silage cover, and management of aerobic deterioration. The possibility of reducing DM and quality losses during the ensiling process requires knowledge of how to measure losses on farm and establish the status of the silage during the feed-out phase, implementing the most effective management practices to avoid air exposure during conservation and reduce silage aerobic deterioration during feeding. The paper concludes with future perspectives and recommended management practices to reduce losses and increase efficiency over the whole ensiling process in view of increasing sustainability of the livestock production chain.

Effect of sex on torque, recovery, EMG, and MMG responses to fatigue.

OBJECTIVE: The purpose of the present investigation was to examine the effect of sex on maximal voluntary isometric contraction (MVIC) torque and the EMG and MMG responses as a result of fatiguing, intermittent, submaximal (65% of MVIC), isometric elbow flexion muscle contractions. METHODS: Eighteen men and women performed MVIC trials before (pretest), after (posttest), and 5-min after (5-min recovery) performing 50 intermittent, submaximal isometric muscle contractions. Surface electromyographic (EMG) and mechanomyographic (MMG) signals were simultaneously recorded from the biceps brachii muscle. RESULTS: As a result of the fatiguing workbout torque decreased similarly from pretest to posttest for both the men (24.0%) and women (23.3%). After 5-min of recovery, torque had partially recovered for the men, while torque had returned to pretest levels for the women. For both sexes, from pretest to posttest EMG mean power frequency and MMG amplitude decreased, but returned to pretest levels after 5-min of recovery. CONCLUSIONS: In the present study, there were sex-related differences in muscle fatigue that were not associated with the EMG or MMG responses.

Effects of fatiguing constant versus alternating intensity intermittent isometric muscle actions on maximal torque and neuromuscular responses.

OBJECTIVE: To determine the effects of constant versus alternating applications of torque during fatiguing, intermittent isometric muscle actions of the leg extensors on maximal voluntary isometric contraction (MVIC) torque and neuromuscular responses. METHODS: Sixteen subjects performed two protocols, each consisting of 50 intermittent isometric muscle actions of the leg extensors with equal average load at a constant 60% MVIC or alternating 40 then 80% (40/80%) MVIC with a work-to-rest ratio of 6-s on and 2-s off. MVIC torque as well as electromyographic signals from the vastus lateralis (VL), vastus medialis (VM), and rectus femoris (RF) and mechanomyographic signals from the VL were recorded pretest, immediately posttest, and 5-min posttest. RESULTS: The results indicated that there were no time-related differences between the 60% MVIC and 40/80% MVIC protocols. The MVIC torque decreased posttest (22 to 26%) and remained depressed 5-min posttest (9%). There were decreases in electromyographic frequency (14 to 19%) and mechanomyographic frequency (23 to 24%) posttest that returned to pretest levels 5-min posttest. There were no changes in electromyographic amplitude and mechanomyogrpahic amplitude. CONCLUSIONS: These findings suggested that these neuromuscular parameters did not track the fatigue-induced changes in MVIC torque after 5-min of recovery.

Four weeks of high- versus low-load resistance training to failure on the rate of torque development, electromechanical delay, and contractile twitch properties.

The purpose of this study was to investigate the effects of 4-weeks of high- versus low-load resistance training to failure on rate of torque development (RTD), electromechanical delay (EMD), and contractile twitch characteristics. Fifteen men (mean±SD; age=21.7±2.4 yrs) were randomly assigned to either a high- (80% 1RM; n=7) or low-load (30% 1RM; n=8) training group and completed elbow flexion resistance training to failure 3 times per week for 4 weeks. The participants were tested at baseline, 2-, and 4-weeks of training. Peak RTD (pRTDV) and RTD at 0-30 (RTD30V), 0-50 (RTD50V), 0-100 (RTD100V), and 0-200 (RTD200V) ms, integrated EMG amplitude (iEMG) at 0-30, 0-50, and 0-100 ms, and EMD were quantified during maximal voluntary isometric muscle actions. Peak twitch torque, peak RTD, time to peak twitch, 1/2 relaxation time and the peak relaxation rate were quantified during evoked twitches. Four weeks of high-load, but not low-load resistance training, increased RTD200V. There were also increases in iEMG during the first 30 ms of muscle activation for the high- and low-load groups, which may have indirectly indicated increases in early phase motor unit recruitment and/or firing frequency. There were no significant training-induced adaptations in EMD or contractile twitch properties.

Electromyographic Responses from the Vastus Medialis during Isometric Muscle Actions.

This study examined the electromyographic (EMG) responses from the vastus medialis (VM) for electrodes placed over and away from the innervation zone (IZ) during a maximal voluntary isometric contraction (MVIC) and sustained, submaximal isometric muscle action. A linear electrode array was placed on the VM to identify the IZ and muscle fiber pennation angle during an MVIC and sustained isometric muscle action at 50% MVIC. EMG amplitude and frequency parameters were determined from 7 bipolar channels of the electrode array, including over the IZ, as well as 10 mm, 20 mm and 30 mm proximal and distal to the IZ. There were no differences between the channels for the patterns of responses for EMG amplitude or mean power frequency during the sustained, submaximal isometric muscle action; however, there were differences between channels during the MVIC. The results of the present study supported the need to standardize the placement of electrodes on the VM for the assessment of EMG amplitude and mean power frequency. Based on the current findings, it is recommended that electrode placements be distal to the IZ and aligned with the muscle fiber pennation angle during MVICs, as well as sustained, submaximal isometric muscle actions.

Physiological Responses during Cycle Ergometry at a Constant Perception of Effort.

13 subjects performed an incremental test to exhaustion, 4, 8-min submaximal rides, and a 1-h ride at the rating of perceived exertion (RPE) that corresponded to the physical working capacity at the OMNI threshold (PWC(OMNI)) to examine: 1) the oxygen consumption (V̇O2), heart rate (HR), minute ventilation (+V̇(E)), respiratory frequency (FR), and power output responses during 1-h work bouts at a constant RPE that corresponded to the PWC(OMNI); and 2) the ability of current models to explain the responses for physiological and perceptual parameters during the 1-h work bouts. The RPE that corresponded to the PWC(OMNI) represented a sustainable exercise intensity (56±5% (V̇O(2Peak)) within the moderate-intensity domain. The mean, normalized slope coefficients for the V̇O2, +V̇(E), and power output vs. time relationships during the 1-h rides were significantly less than zero. The mean, normalized slope coefficient for the FR vs. time relationship during the 1-h rides, however, was not significantly different from zero. Thus, RPE most clearly tracked FR responses during the 1-h rides. It was hypothesized that afferent feedback from respiratory muscles may have mediated the perception of effort during cycle ergometry at a constant RPE in the moderate-intensity domain.

The rate of torque development: a unique, non-invasive indicator of eccentric-induced muscle damage?

This study examined the time courses of recovery for isometric peak torque and rate of torque development (RTD) after eccentric-induced muscle damage. 18 men completed 6 sets of 10 maximal eccentric isokinetic muscle actions at 30° · s(-1). Peak torque, peak RTD and RTD at 10 (RTD10), 50 (RTD50), 100 (RTD100) and 200 ms (RTD200), serum creatine kinase and lactate dehydrogenase were measured before (PRE), immediately after (POST), 24, 48 and 72 h after eccentric exercise. Creatine kinase and lactate dehydrogenase increased from 139 to 6 457 and from 116 to 199 IU · L(-1) from PRE to 72 h, respectively. Peak torque and all RTDs decreased at POST. Peak torque and RTD200 remained lower than PRE through 72 h. Peak RTD remained lower than PRE through 48 h, but was not different from PRE at 72 h. RTD10 and RTD100 were lower than PRE through 24 h, but were not different from PRE at 48 and 72 h. RTD50 decreased at POST, but was not different from PRE at 24 h. Early phase RTDs recovered more quickly than PT and RTD200. Early phase RTDs may reflect neural mechanisms underlying eccentric-induced force decrements, while late RTDs may describe the same physiological mechanisms as PT.

Effect of physical damage to ears of corn before harvest and treatment with various additives on the concentration of mycotoxins, silage fermentation, and aerobic stability of corn silage.

We studied the effects of damaging ears of corn in the field prior to harvest and the use of various additives on the production of selected mycotoxins, silage fermentation, and aerobic stability of whole plant corn. In experiment 1, ears of corn were undamaged or were slashed with a knife 7 d before harvesting, exposing damaged kernels to the environment. Corn plants were harvested (about 35% DM) and treated in a 2 × 2 factorial arrangement of treatments. Treatments were undamaged or damaged plants, untreated or treated with Lactobacillus buchneri 40788 (400,000 cfu/g of fresh forage) and Pediococcus pentosaceus (100,000 cfu/g). Damaging ears prior to harvest increased the amount of fumonisin but decreased the amount of starch in harvested corn plants. After ensiling, corn silage made from plants damaged before harvest had lower starch but greater concentrations of deoxynivalenol and fumonisin than silage made from plants that were undamaged. Microbial inoculation resulted in fewer yeasts and lower concentrations of zearalenone in silage when compared to uninoculated silage. Inoculated silage also had more acetic acid and 1,2-propanediol than did uninoculated silage. In experiment 2, ears of corn were undamaged or were slashed with a knife 27 d or 9 d before harvesting for corn silage. Whole plants were harvested at about 36% DM in a 2 × 3 factorial arrangement of treatments. Factors were time of damaging the ears (27 d, 9 d, or no damage) relative to harvest and no additive or 0.1% (fresh weight) potassium sorbate. Damaging plants 9 d prior to harvest did not affect the concentrations of deoxynivalenol, fumonisin, and zearalenone in plants at harvest. However, concentrations of deoxynivalenol and fumonisin were increased in fresh forage that had ears damaged at 27 d when compared to corn plants that were undamaged. Corn plants damaged for 27 d prior to harvest also had a lower concentration of starch than corn damaged for 9 d but was higher in acid detergent fiber than other treatments. The addition of potassium sorbate at harvest had no effect on the concentrations of mycotoxins in the resulting silage, but concentrations of mycotoxins were still greatest in silage made from plants that were damaged the longest prior to harvest (27 d). Silages treated with potassium sorbate had fewer yeasts and molds than silages without the additive. Damaging ears of corn before harvest had no effects on the aerobic stability of silages in both experiments. In contrast, the addition of the inoculant and potassium sorbate improved aerobic stabilities of silages when compared to untreated silages. These studies showed that physical damage to ears of corn prior to harvest can result in the production of mycotoxins in the field. This finding suggests that producers should test corn silage for mycotoxins prior to feeding especially if the forage has been subjected to physical damage prior to ensiling.

Influence of ensiling time and inoculation on alteration of the starch-protein matrix in high-moisture corn.

The fates of hydrophobic zein proteins, which encapsulate corn starch to create vitreous endosperm, have not been investigated in high-moisture corn (HMC). To assess influences of ensiling time and inoculation on zein proteins in HMC, quadruplicate samples of 2 random corn hybrids (A and B), containing 25.7 and 29.3% moisture, were ground, inoculated with (I) or without 600,000 cfu/g of Lactobacillus buchneri 40788 (Lallemand Animal Nutrition, Milwaukee, WI), and ensiled for 0, 15, 30, 60, 120, and 240 d. Nutrient composition [crude protein (CP), starch, acid detergent fiber, and neutral detergent fiber], fermentation (pH, lactate, and acetate), and protein degradation markers (buffer-soluble CP, isopropanol-soluble CP, and NH(3)-N) were evaluated. At 0 and 240 d, α, γ, δ, and ß zein subunits were profiled using HPLC. Data were evaluated as a split-split plot using the PROC MIXED procedures of SAS. Ensiling time and inoculation decreased pH, and altered lactate and acetate contents of HMC. Lactate and acetate contents of A, AI, B, and BI at 240 d were 0.40, 0.32, 1.11, 0.73, and 0, 0.35, 0.30, and 0.87% of DM, respectively. Buffer-soluble CP in HMC increased from 1.5 to 2.0% of DM at 0 d to >4.0% of DM at 240 d. Inoculation had no effect on buffer-soluble CP but increased NH(3)-N content of HMC. Corn A contained more isopropanol-soluble CP than did corn B and peak areas for 6 α, and all γ and δ zein regions were greater for corn A. Ensiling (0 vs. 240 d) decreased all zein subunits with the exception of 2 α and 1 δ subunit. Ensiling decreased (42.2-73.2%) γ zeins, which are primarily responsible for cross-linking in the starch-protein matrix. Despite altering lactate and acetate contents, inoculation had no effect on degrading hydrophobic zein proteins in HMC. Data suggest that hydrophobic zein proteins in the starch-protein matrix of HMC are degraded by proteolytic activity over an extended ensiling time.

The effects of Lactobacillus buchneri with or without a homolactic bacterium on the fermentation and aerobic stability of corn silages made at different locations.

Whole-plant corn (31 to 39% dry matter) from several locations was chopped, treated with nothing (U), Lactobacillus buchneri 40788 (4 x 10(5) cfu/g; LB), or L. buchneri (4 x 10(5) cfu/g) and Pediococcus pentosaceus (1 x 10(5) cfu/g; LBPP), and packed into quadruplicate 20-L silos to determine their effects on silage fermentation and aerobic stability after 120 d of storage. The experiment was a randomized complete block design with main effects of treatment (T), block (location; L), and T x L interaction. Dry matter recovery was different among locations but unaffected by T. The population of lactic acid bacteria was greater in LB and LBPP than in U, and the opposite was true regarding the population of yeasts. Numbers of L. buchneri (colony-forming unit equivalents), determined by a real-time quantitative polymerase chain reaction, were higher in 4 of 5 locations for LB and LBPP compared with U (T x L interaction) with an average 6.70 log cfu/g for LB and LBPP versus 4.87 log cfu/g for U. Silages inoculated with LB and LBPP had higher silage pH and higher concentrations of acetic acid and 1,2 propanediol but lower concentrations of ethanol and water-soluble carbohydrates; there was a T x L interaction for all these variables. Aerobic stability was improved by LB and LBPP (mean of 136 h) compared with U (44 h), but there was an interaction between T x L. In general, locations with the highest population of L. buchneri had the largest increases in acetic acid and, consequently, the greatest improvements in aerobic stability. The addition of L. buchneri 40788 alone or with P. pentosaceus resulted in similar effects on silage fermentation and aerobic stability, but the effects were variable among locations, suggesting that unidentified factors; for example, in the field or on the forage crop, may alter the effectiveness of microbial inoculation.

The effect of wide swathing on wilting times and nutritive value of alfalfa haylage.

On 3 consecutive cuttings, alfalfa from a single field was mowed with a John Deere 946 mower-conditioner (4-m cut width; Moline, IL) to leave narrow swaths (NS) ranging from 1.2 to 1.52 m wide (30-37% of cutter bar width) and wide swaths (WS) ranging from 2.44 to 2.74 m wide (62-67% of cutter bar width). Samples were collected from windrows and dry matter (DM) was monitored during wilting until a target of 43 to 45% DM was obtained. Forage from random windrows (n=4-6) was harvested by hand, chopped through a forage harvester before being packed in replicated vacuum-sealed bags, and allowed to ensile for 65 d. There was no swath width x cutting interaction for any parameter tested. Over all cuttings, the resulting silage DM was not different between the NS silage (43.8%) and the WS silage (44.9%). However, wide swathing greatly reduced the time of wilting before making silage. The hours of wilting time needed to reach the targeted DM for the NS silage compared with the WS silage at cuttings 1, 2, and 3 were 50 versus 29, 54 versus 28, and 25 versus 6, respectively. At the time of ensiling, the WS silage had more water-soluble carbohydrates (5.1%) than did the NS silage (3.7%). The WS silage had a lower pH (4.58) than did the NS silage (4.66), but swath width did not affect fermentation end products (lactic acid, acetic acid, and ethanol). The NS silage had more NH(3)-N (0.26%) than did the WS silage (0.21%). Wide swathing did not affect the concentration of ash or the digestibility of NDF, but it lowered the N content (NS=3.45%; WS=3.23%) and increased the ADF content (NS=39.7%; WS=40.9%) of the resulting silage. Wide swathing can markedly reduce the time that alfalfa must wilt before it can be chopped for silage, but under good conditions, as in this study, the resulting silage quality was generally not improved.

A mechanomyographic fatigue threshold test for cycling.

The purposes of this study were twofold: 1) to derive the mechanomyographic mean power frequency fatigue threshold (MMG MPFFT) for submaximal cycle ergometry; and 2) to compare the power outputs associated to the MMG MPFFT to other neuromuscular and gas exchange fatigue thresholds. 9 adults (5 men and 4 women; mean+/-SD age=23.7+/-3.7 years; body weight=66.3+/-8.2 kg) performed an incremental cycle ergometry test to exhaustion while expired gas samples, electromyographic (EMG), and MMG signals were measured from the vastus lateralis muscle. The non-significant correlations (r=0.17 to 0.66; p>0.05) among the physical working capacity at the fatigue threshold (PWCFT), MMG MPFFT, and gas exchange threshold (GET) suggested that different physiological mechanisms may underlie these 3 fatigue thresholds. A significant correlation (r=0.83) for the MPFFT vs. respiratory compensation point (RCP) suggested that these fatigue thresholds may be mediated by a common physiological mechanism. In addition, the significantly lower mean values found for the PWCFT (mean+/-SD=163+/-43 W), MMG MPFFT (132+/-33 W), and GET (144+/-28 W) than MPFFT (196+/-53 W) and RCP (202+/-41 W) suggested that these gas exchange and neuromuscular fatigue thresholds may demarcate different exercise intensity domains.

The effects of parallel versus perpendicular electrode orientations on EMG amplitude and mean power frequency from the biceps brachii.

The purposes of this study were threefold: (1) to compare the isometric torque-related patterns of absolute and normalized electromyographic (EMG) amplitude and mean power frequency (MPF) responses for electrode orientations that were parallel and perpendicular to the muscle fibers; (2) to examine the influence of electrode orientation on mean absolute EMG amplitude and MPF values; and (3) to determine the effects of normalization on mean EMG amplitude and MPF values from parallel and perpendicular electrode orientations. Ten adults (5 men and 5 women mean +/- SD age = 23.8 +/- 2.3 years) volunteered to participate in the investigation. Two sets of bipolar surface EMG electrodes (20 mm center to center) were placed parallel and perpendicular to the muscle fibers over the biceps brachii. The subjects performed a maximal voluntary isometric contraction (MVIC) test followed by randomly ordered submaximal muscle actions in 10% increments from 10 to 90% MVIC. Paired t-tests indicated that absolute EMG amplitude values for the parallel electrode orientation were greater (p < 0.05) than those for the perpendicular orientation at all isometric torque levels except 10% MVIC For normalized EMG amplitude values, however, there were no significant mean differences between electrode orientations. There were also no differences between electrode orientations for absolute or normalized EMG MPF values. In 30% of the cases, different torque-related patterns of responses were observed between the parallel and perpendicular electrode orientations for the absolute and normalized EMG amplitude and MPF values. Therefore, the results of the present study support the need for standardizing electrode orientation to compare the pattern of responses for EMG amplitude and MPF values and normalizing EMG amplitude data to compare the mean values.

The effect of epoch length on the electromyographic mean power frequency and amplitude versus time relationships.

UNLABELLED: The electromyographic (EMG) mean power frequency (MPF) and amplitude versus time relationships are commonly used to characterize localized muscle fatigue. PURPOSE: The purpose of this study was to compare the effect of epoch length on the individual and mean slope coefficients and y-intercepts resulting from the EMG MPF and amplitude versus time relationships of the vastus medialis (VM) muscle during fatiguing isometric muscle actions at 30 and 75% of maximum voluntary isometric contraction (MVC). METHODS: Eight adults performed two continuous, isometric muscle actions of the leg extensors at 30 and 75% MVC to exhaustion. Six, 5.0 s epochs of the surface EMG signals were recorded from the VM during each minute. Epoch lengths of 0.5, 1.0, and 2.0 s were selected from the middle of each 5.0 s epoch. Linear regression was used to estimate the slope coefficient and y-intercept values for the EMG MPF and amplitude versus time relationships for each epoch length (0.5, 1.0, 2.0, and 5.0 s) and subject. RESULTS: There were no significant differences between epoch lengths for the individual or mean slope coefficients or y-intercepts (EMG MPF and amplitude versus time relationships). CONCLUSION: This study indicated that epochs of 0.5 - 5.0 s resulted in the same characterization of EMG (MPF and amplitude) versus time relationships during isometric muscle actions.

Sites of synthesis of chloroplast ribosomal proteins in Chlamydomonas.

Cells of Chlamydomonas reinhardtii were pulse-labeled in vivo in the presence of inhibitors of cytoplasmic (anisomycin) or chloroplast (lincomycin) protein synthesis to ascertain the sites of synthesis of chloroplast ribosomal proteins. Fluorographs of the labeled proteins, resolved on two-dimensional (2-D) charge/SDS and one-dimensional (1-D) SDS-urea gradient gels, demonstrated that five to six of the large subunit proteins are products of chloroplast protein synthesis while 26 to 27 of the large subunit proteins are synthesized on cytoplasmic ribosomes. Similarly, 14 of 31 small subunit proteins are products of chloroplast protein synthesis, while the remainder are synthesized in the cytoplasm. The 20 ribosomal proteins shown to be made in the chloroplast of Chlamydomonas more than double the number of proteins known to be synthesized in the chloroplast of this alga.

Chloroplast ribosomal proteins of Chlamydomonas synthesized in the cytoplasm are made as precursors.

Polyadenylated RNA from Chlamydomonas was translated in a cell-free rabbit reticulocyte system that employed [35S]methionine. Antibodies made to four chloroplast ribosomal proteins synthesized in the cytoplasm and imported into the organelle were used for indirect immunoprecipitation of the labeled translation products, which were subsequently visualized on fluorographs of SDS gels. The cytoplasmically synthesized chloroplast ribosomal proteins were first seen as precursors with apparent molecular weights of 1,000 to 6,000 greater than their respective mature forms. Processing of the ribosomal protein precursors to mature proteins was affected by adding a postribosomal supernatant that had been extracted from cells of Chlamydomonas. In contrast to the chloroplast ribosomal proteins synthesized in the cytoplasm, two such proteins made within the chloroplast were found to be synthesized in mature form in cell-free wheat germ translation systems programmed with nonpolyadenylated RNA.

Transposon tagging and molecular analysis of the maize regulatory locus opaque-2.

Genetic analyses suggested that the opaque-2 (o2) locus in maize acts as a positive, transacting, transcriptional activator of the zein seed storage-protein genes. Because isolation of the gene is requisite to understanding the molecular details of this regulation, transposon mutagenesis with the transposable element suppressor-mutator (Spm) was carried out, and three mutable o2 alleles were obtained. One of these alleles contained an 8.3-kilobase autonomous Spm, another a 6.8-kilobase nonautonomous Spm, and the third an unidentified transposon that is unrelated to Spm. A DNA sequence flanking the autonomous Spm insertion was verified to be o2-specific and provided a probe to clone a wild-type allele. Northern blots indicated that the gene is expressed in wild-type endosperm but not in leaf tissues or in endosperms homozygous for a mutant allele of the O2 gene. A transcript was detected in endosperms homozygous for mutations at opaque-7 and floury-2, an indication that O2 expression is independent of these two other putative regulators of zein synthesis.

Diversification of C-function activity in maize flower development.

The Arabidopsis gene AGAMOUS is required for male and female reproductive organ development and for floral determinacy. Reverse genetics allowed the isolation of a transposon-induced mutation in ZAG1, the maize homolog of AGAMOUS. ZAG1 mutants exhibited a loss of determinacy, but the identity of reproductive organs was largely unaffected. This suggested a redundancy in maize sex organ specification that led to the identification and cloning of a second AGAMOUS homolog, ZMM2, that has a pattern of expression distinct from that of ZAG1. C-function organ identity in maize (as defined by the A, B, C model of floral organ development) may therefore be orchestrated by two closely related genes, ZAG1 and ZMM2, with overlapping but nonidentical activities.

The development of lactic acid bacteria and Lactobacillus buchneri and their effects on the fermentation of alfalfa silage.

This study was conducted to document the development of populations of lactic acid bacteria (LAB) and Lactobacillus buchneri in alfalfa silage treated with various inoculants. Wilted and chopped alfalfa (45% dry matter) was treated with 1) distilled water (untreated, U), 2) Lactobacillus buchneri 40788 (4 x 10(5) cfu/g; LB), or 3) L. buchneri 40788 (4 x 10(5) cfu/g) and Pediococcus pentosaceus (1 x 10(5) cfu/g; LBPP). Forages were packed into triplicate vacuum-sealed, nylon-polyethylene bags per treatment, and ensiled for 2, 5, 45, 90, and 180 d. Viable (cfu) LAB in forage and silage were quantified by traditional plating on selective agar, and numbers of L. buchneri (cfu-equivalent, cfu-E) were quantified by real-time quantitative PCR. Fresh, untreated forage had 5.52 log cfu of LAB/g and 3.79 log cfu-E of L. buchneri/g. After 2 d of ensiling, numbers of LAB increased to >8 log cfu/g in all silages. In contrast, numbers of L. buchneri in U remained below 4 log cfu-E/g but reached approximately 7 log cfu-E/g in LB and LBPP. From d 5 onward, numbers of L. buchneri in U remained below 6 log cfu-E/g but approached 9 log cfu-E/g in LB and LBPP. The pH was lower in LBPP compared with U and LB after 2 and 5 d of ensiling, but pH was lower for U compared with LB and LBPP thereafter. Treatments LB and LBPP had more acetic acid than U at 45 d of ensiling, which coincided with detectable amounts of 1,2 propanediol. Inoculation with LBPP resulted in silage with the highest concentration of 1,2 propanediol after 180 d of ensiling. From d 45 onward, LB and LBPP silages had lower concentrations of residual water-soluble carbohydrates but had higher concentrations of ammonia-N than U. In conclusion, epiphytic L. buchneri can be detected in alfalfa but this population is unable to lead the silage fermentation. In contrast, when L. buchneri was added to silage as an inoculant, the numbers of L. buchneri (cfu-E) increased markedly but did not dictate fermentation until 45 d of ensiling. These findings help to explain why the response (in increased acetic acid) from the addition of L. buchneri in silages is not immediate.