U.S. Dairy Forage Research Center

Fundamental features of microbial cellulose utilization are examined at successively higher levels of aggregation encompassing the structure and composition of cellulosic biomass, taxonomic diversity, cellulase enzyme systems, molecular biology of cellulase enzymes, physiology of cellulolytic microorganisms, ecological aspects of cellulase-degrading communities, and rate-limiting factors in nature. The methodological basis for studying microbial cellulose utilization is considered relative to quantification of cells and enzymes in the presence of solid substrates as well as apparatus and analysis for cellulose-grown continuous cultures. Quantitative description of cellulose hydrolysis is addressed with respect to adsorption of cellulase enzymes, rates of enzymatic hydrolysis, bioenergetics of microbial cellulose utilization, kinetics of microbial cellulose utilization, and contrasting features compared to soluble substrate kinetics. A biological perspective on processing cellulosic biomass is presented, including features of pretreated substrates and alternative process configurations. Organism development is considered for "consolidated bioprocessing" (CBP), in which the production of cellulolytic enzymes, hydrolysis of biomass, and fermentation of resulting sugars to desired products occur in one step. Two organism development strategies for CBP are examined: (i) improve product yield and tolerance in microorganisms able to utilize cellulose, or (ii) express a heterologous system for cellulose hydrolysis and utilization in microorganisms that exhibit high product yield and tolerance. A concluding discussion identifies unresolved issues pertaining to microbial cellulose utilization, suggests approaches by which such issues might be resolved, and contrasts a microbially oriented cellulose hydrolysis paradigm to the more conventional enzymatically oriented paradigm in both fundamental and applied contexts.

The lignin biosynthetic pathway has been studied for more than a century but has undergone major revisions over the past decade. Significant progress has been made in cloning new genes by genetic and combined bioinformatics and biochemistry approaches. In vitro enzymatic assays and detailed analyses of mutants and transgenic plants altered in the expression of lignin biosynthesis genes have provided a solid basis for redrawing the monolignol biosynthetic pathway, and structural analyses have shown that plant cell walls can tolerate large variations in lignin content and structure. In some cases, the potential value for agriculture of transgenic plants with modified lignin structure has been demonstrated. This review presents a current picture of monolignol biosynthesis, polymerization, and lignin structure.

The Cornell Net Carbohydrate and Protein System (CNCPS) has a submodel that predicts rates of feedstuff degradation in the rumen, the passage of undegraded feed to the lower gut, and the amount of ME and protein that is available to the animal. In the CNCPS, structural carbohydrate (SC) and nonstructural carbohydrate (NSC) are estimated from sequential NDF analyses of the feed. Data from the literature are used to predict fractional rates of SC and NSC degradation. Crude protein is partitioned into five fractions. Fraction A is NPN, which is trichloroacetic (TCA) acid-soluble N. Unavailable or protein bound to cell wall (Fraction C) is derived from acid detergent insoluble nitrogen (ADIP), and slowly degraded true protein (Fraction B3) is neutral detergent insoluble nitrogen (NDIP) minus Fraction C. Rapidly degraded true protein (Fraction B1) is TCA-precipitable protein from the buffer-soluble protein minus NPN. True protein with an intermediate degradation rate (Fraction B2) is the remaining N. Protein degradation rates are estimated by an in vitro procedure that uses Streptomyces griseus protease, and a curve-peeling technique is used to identify rates for each fraction. The amount of carbohydrate or N that is digested in the rumen is determined by the relative rates of degradation and passage. Ruminal passage rates are a function of DMI, particle size, bulk density, and the type of feed that is consumed (e.g., forage vs cereal grain).

As an important constituent of animal feeds, fiber represents the portion of feeds that is bulky and difficult to digest. The neutral detergent fiber (NDF) method, developed over 30 years ago, is the method of choice for measuring total fiber in forages and other feeds. Several modifications that were made to improve its general applicability to all feeds and others developed in individual laboratories often resulted in variability among laboratories in measuring NDF. The amylase-treated NDF (aNDF) method, therefore, was developed as an accurate and precise method of measuring total insoluble fiber in feeds. A collaborative study was conducted to evaluate the repeatability and reproducibility of the aNDF method over the full range of animal feed materials. Twelve laboratories representing research, feed company, regulatory, and commercial feed testing laboratories analyzed 11 materials as blind duplicates. The materials represented feed matrixes, including animal products; high-protein, high-fat, and high-pectin feeds; oil seeds; grains; heated by-product feeds; and legume and grass hays and silages. Materials selected varied in chemical composition and contained 0-90% aNDF, 1-16% ash, 1-20% crude fat, 1-40% crude protein, and 0-50% starch. Correcting results for changes in blanks and reporting results as ash-free aNDF organic matter (aNDFom) improved the repeatability and reproducibility of results when aNDF was <25%. The within-laboratory repeatability standard deviation (Sr) for percentage aNDFom in feeds varied from 0.21 to 1.82 and among-laboratory reproducibility standard deviation (S(R)) varied from 0.37 to 2.24. The HORRAT was <2 for all materials except feed materials containing >10% fat. However, standard deviations of repeatability and reproducibility for feeds with >10% fat were similar to those of other materials. It is recommended that the aNDF method be accepted for Official First Action status.

Current NRC recommendations for dairy cattle provide limited guidance to nutritionists for meeting the fiber and carbohydrate needs of lactating cows. The NRC provide only minimum recommendations for fiber and no accommodation for factors such as physical effectiveness of fiber, interactions with nonfibrous carbohydrates, or animal attributes, which can affect the optimality of dairy rations. To be an improvement, any new system for meeting the fiber requirements of dairy cows must be based on 1) feed characteristics that can be defined and preferably be determined quantitatively using routine laboratory methods and 2) animal requirements that correspond to critical feed characteristics and vary with feeding situation, ration composition, and attributes of the animal. Published data were used to develop coefficients for defining the physical effectiveness or roughage value of feeds and the fiber requirements of dairy cows. Information in this paper is intended to provide practical guidelines for improving current fiber recommendations and to serve as an idealized framework for future research on meeting the fiber requirements of dairy cows. The system is based on NDF as the measure of total chemical fiber in feeds. Adjustments for the effectiveness of NDF in maintaining milk fat production and optimizing ruminal fermentation are based on the particle size and inherent characteristics of NDF that affect chewing activity, ruminal pH, and milk fat production.

The Cornell Net Carbohydrate and Protein System (CNCPS) has a kinetic submodel that predicts ruminal fermentation. The ruminal microbial population is divided into bacteria that ferment structural carbohydrate (SO and those that ferment nonstructural carbohydrate (NSC). Protozoa are accommodated by a decrease in the theoretical maximum growth yield (.50 vs .40 g of cells per gram of carbohydrate fermented), and the yields are adjusted for maintenance requirements (.05 vs .150 g of cell dry weight per gram of carbohydrate fermented per hour for SC and NSC bacteria, respectively). Bacterial yield is decreased when forage NDF is < 20% (2.5% for every 1% decrease in NDF). The SC bacteria utilize only ammonia as a N source, but the NSC bacteria can utilize either ammonia or peptides. The yield of NSC bacteria is enhanced by as much as 18.7% when proteins or peptides are available. The NSC bacteria produce less ammonia when the carbohydrate fermentation (growth) rate is rapid, but 34% of the ammonia production is insensitive to the rate of carbohydrate fermentation. Ammonia production rates are moderated by the rate of peptide and amino acid uptake (.07 g of peptide per gram of cells per hour), and peptides and amino acids can pass out of the rumen if the rate of proteolysis is faster than the rate of peptide utilization. The protein-sparing effect of ionophores is accommodated by decreasing the rate of peptide uptake by 34%. Validation with published data of microbial flow from the rumen gave a regression with a slope of .94 and an r2 of .88.

Additives have been available for enhancing silage preservation for decades. This review covers research studies published since 2000 that have investigated the efficacy of silage additives. The review has been divided into 6 categories of additives: homofermentative lactic acid bacteria (LAB), obligate heterofermentative LAB, combination inoculants containing obligate heterofermentative LAB plus homofermentative LAB, other inoculants, chemicals, and enzymes. The homofermentative LAB rapidly decrease pH and increase lactic acid relative to other fermentation products, although a meta-analysis indicated no reduction in pH in corn, sorghum, and sugarcane silages relative to untreated silages. These additives resulted in higher milk production according to the meta-analysis by mechanisms that are still unclear. Lactobacillus buchneri is the dominant species used in obligate heterofermentative LAB silage additives. It slowly converts lactic acid to acetic acid and 1,2-propanediol during silo storage, improving aerobic stability while having no effect on animal productivity. Current research is focused on finding other species in the Lb. buchneri group capable of producing more rapid improvements in aerobic stability. Combination inoculants aim to provide the aerobic stability benefits of Lb. buchneri with the silage fermentation efficiency and animal productivity benefits of homofermentative LAB. Research indicates that these products are improving aerobic stability, but feeding studies are not yet sufficient to make conclusions about effects on animal performance. Novel non-LAB species have been studied as potential silage inoculants. Streptococcus bovis is a potential starter species within a homofermentative LAB inoculant. Propionibacterium and Bacillus species offer improved aerobic stability in some cases. Some yeast research has focused on inhibiting molds and other detrimental silage microorganisms, whereas other yeast research suggests that it may be possible to apply a direct-fed microbial strain at ensiling, have it survive ensiling, and multiply during feed out. Chemical additives traditionally have fallen in 2 groups. Formic acid causes direct acidification, suppressing clostridia and other undesired bacteria and improving protein preservation during ensiling. On the other hand, sorbic, benzoic, propionic, and acetic acids improve silage aerobic stability at feed out through direct inhibition of yeasts and molds. Current research has focused on various combinations of these chemicals to improve both aerobic stability and animal productivity. Enzyme additives have been added to forage primarily to breakdown plant cell walls at ensiling to improve silage fermentation by providing sugars for the LAB and to enhance the nutritive value of silage by increasing the digestibility of cell walls. Cellulase or hemicellulase mixtures have been more successful at the former than the latter. A new approach focused on Lb. buchneri producing ferulic acid esterase has also had mixed success in improving the efficiency of silage digestion. Another new enzyme approach is the application of proteases to corn silage to improve starch digestibility, but more research is needed to determine the feasibility. Future silage additives are expected to directly inhibit clostridia and other detrimental microorganisms, mitigate high mycotoxin levels on harvested forages during ensiling, enhance aerobic stability, improve cell wall digestibility, increase the efficiency of utilization of silage nitrogen by cattle, and increase the availability of starch to cattle.

Many opportunities exist to reduce enteric methane (CH4) and other greenhouse gas (GHG) emissions per unit of product from ruminant livestock. Research over the past century in genetics, animal health, microbiology, nutrition, and physiology has led to improvements in dairy production where intensively managed farms have GHG emissions as low as 1 kg of CO2 equivalents (CO2e)/kg of energy-corrected milk (ECM), compared with >7 kg of CO2e/kg of ECM in extensive systems. The objectives of this review are to evaluate options that have been demonstrated to mitigate enteric CH4 emissions per unit of ECM (CH4/ECM) from dairy cattle on a quantitative basis and in a sustained manner and to integrate approaches in genetics, feeding and nutrition, physiology, and health to emphasize why herd productivity, not individual animal productivity, is important to environmental sustainability. A nutrition model based on carbohydrate digestion was used to evaluate the effect of feeding and nutrition strategies on CH4/ECM, and a meta-analysis was conducted to quantify the effects of lipid supplementation on CH4/ECM. A second model combining herd structure dynamics and production level was used to estimate the effect of genetic and management strategies that increase milk yield and reduce culling on CH4/ECM. Some of these approaches discussed require further research, but many could be implemented now. Past efforts in CH4 mitigation have largely focused on identifying and evaluating CH4 mitigation approaches based on nutrition, feeding, and modifications of rumen function. Nutrition and feeding approaches may be able to reduce CH4/ECM by 2.5 to 15%, whereas rumen modifiers have had very little success in terms of sustained CH4 reductions without compromising milk production. More significant reductions of 15 to 30% CH4/ECM can be achieved by combinations of genetic and management approaches, including improvements in heat abatement, disease and fertility management, performance-enhancing technologies, and facility design to increase feed efficiency and life-time productivity of individual animals and herds. Many of the approaches discussed are only partially additive, and all approaches to reducing enteric CH4 emissions should consider the economic impacts on farm profitability and the relationships between enteric CH4 and other GHG.

BACKGROUND: Major advances in selection progress for cattle have been made following the introduction of genomic tools over the past 10-12 years. These tools depend upon the Bos taurus reference genome (UMD3.1.1), which was created using now-outdated technologies and is hindered by a variety of deficiencies and inaccuracies. RESULTS: We present the new reference genome for cattle, ARS-UCD1.2, based on the same animal as the original to facilitate transfer and interpretation of results obtained from the earlier version, but applying a combination of modern technologies in a de novo assembly to increase continuity, accuracy, and completeness. The assembly includes 2.7 Gb and is >250× more continuous than the original assembly, with contig N50 >25 Mb and L50 of 32. We also greatly expanded supporting RNA-based data for annotation that identifies 30,396 total genes (21,039 protein coding). The new reference assembly is accessible in annotated form for public use. CONCLUSIONS: We demonstrate that improved continuity of assembled sequence warrants the adoption of ARS-UCD1.2 as the new cattle reference genome and that increased assembly accuracy will benefit future research on this species.

Plasma NEFA concentrations increase prior to and at parturition, resulting in increased fatty acid uptake by the liver, fatty acid esterification, and triglyceride storage. Liver triglyceride concentration increases four- to fivefold between d 17 prior to calving and d 1 following calving. Increases in liver triglyceride following calving do not appear to be dramatic. Severity of fatty liver 1 d postpartum is correlated negatively with feed intake 1 d prepartum. Export of newly synthesized triglyceride as very low density lipoprotein occurs slowly in ruminants and is a major factor in the development of fatty liver. Nutritional strategies to minimize the elevation in plasma NEFA prior to calving results in lower liver triglyceride at calving. Fatty liver probably precedes clinical spontaneous ketosis. Liver triglyceride to glycogen ratio may be used to predict susceptibility of cows to ketosis. Consequently, strategies to reduce liver triglyceride at calving may decrease incidence of ketosis. Research to determine methods to reduce fatty acid delivery to the liver or to enhance hepatic export of very low density lipoprotein near calving is warranted. Identification of the cause for the slow rate of assembly and secretion of hepatic very low density lipoprotein in ruminants will be required to assess the feasibility of increasing export of very low density lipoprotein.

In many parts of the world, conserved forage is an essential component of ruminant diets during those intervals when fresh crops are unavailable. A. J. G. Barnett subdivided the process of ensiling into four principal phases of different length and intensity, which cannot be separated precisely from each other. When silage is used as a feed source on a farm, several events take place that have relevance to the number of clostridial spores in silage and milk. During mowing and harvesting of a silage crop, contamination of the crop by soil particles, which contain clostridial spores, is unavoidable. The microbial population of standing or freshly harvested forage crops is considerably different from that found during the process of silage fermentation or in the final product. Most inoculants consist exclusively of homofermentative or facultatively heterofermentative lactic acid bacteria because these are the most efficient producers of lactic acid.

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.

Even under the intensive concentrate feeding systems of ruminant animal production in the United States, forages continue to represent the single most important feed resource. Cell-wall concentration and digestibility limit the intake potential and energy availability of forage crops in beef and dairy production. Identification of cell-wall characteristics that should be targets of genetic modification is required if plant breeders and molecular biologists are to successfully improve forages for livestock feeding. As the forage plant cell develops, phenolic acids and lignin are deposited in the maturing cell wall in specific structural conformations, and in a strict developmental sequence. Lignin is the key element that limits cell-wall digestibility, but cross-linkage of lignin and wall polysaccharides by ferulic acid bridges may be a prerequisite for lignin to exert its affect. Lignin composition and p-coumaric acid in the wall are less likely to affect digestibility. Voluntary intake of forages is a critical determinant of animal performance and cell-wall concentration is negatively related to intake of ruminants consuming high-forage diets. Cell walls affect intake by contributing to ruminal fill. A simple model of cell-wall digestion and passage in which ruminal fill is a function of rates of digestion and passage, as well as the indigestible fraction of the cell-wall indicates that cell-wall concentration and rate of passage are the most critical parameters determining ruminal fill. Plant factors that affect rate of passage include those that affect particle size reduction by chewing and those that affect particle buoyancy in the rumen. The latter is primarily affected by 1) the ability of the particulate matter to retain gases, which is probably related to plant anatomy and rate of digestion of the plant tissue, and 2) the rate at which the gas is produced, which is affected by the potentially digestible fraction of the particulate matter and the rate of digestion of this fraction. Increasing rate of digestion should increase rate of passage by diminishing the gas produced and increasing density over time. A reduction in the indigestible cell-wall fraction is beneficial because this will decrease fill and increase digestibility. Animal production and economic benefits from reduced cell-wall concentration and increased digestibility are significant. Because of the high cell-wall concentration and large digestible cell-wall fraction of grasses, reduction in cell-wall concentration would probably be of greater value than improving digestibility in these species. Legumes represent the opposite situation and may benefit more from improvements in the digestibility of their cell walls.

This chapter reviews the mechanisms of intake regulation, presents integrated models of intake concepts that include both animal and dietary factors, and discusses variation in intake and factors affecting the measurement of forage intake potential. It focuses on the non-forage factors that influence or regulate intake for the purpose of providing background information that will be useful in interpreting intake as a component of forage quality. The effect of nutrient deficiencies on intake has been discussed by J. M. Forbes. The chapter also presents the concepts of intake regulation that indicate the complexity of intake measurement and the role that animals play in determining intake in any given situation. Some scientists are critical of the concept that only physical fill limits the intake of low energy, high fill diets and that only chemical factors or physiological energy demand regulates the intake of high energy, low fill diets for being too simplistic.

Intake and digestibility of feeds by ruminants are influenced by characteristics of the feed, animal and feeding situation. Integration of these characteristics in mathematical models is critical to future progress in forage evaluation and optimal formulation of diets for ruminants. The physiological and physical theories of intake regulation can be described by simple mathematical equations. These equations indicate that intake is a linear function of animal characteristics, such as body weight and production level, and a reciprocal function of feed characteristics, such as fill effect and energy content. Theoretical equations were developed to predict intake when the neutral detergent fiber and energy content of the diet and the energy requirements of the animal are known. The theoretical model also can be used to predict the maximum intake that will maintain a given level of animal production by solving the physiological and physical intake equations at their intersection. Psychogenic intake regulation, which is related to the animal's behavioral response to factors not related to physiological or physical characteristics, can be described mathematically as a multiplier. Digestibility can be predicted by summing the contents of ideal nutritive entities in feeds, which have true digestibilities near 100%, subtracting their associated endogenous losses and adding the variable digestible fiber content. Steady-state models indicate fractional rates of digestion and passage can be used to define ideal nutritive entities and predict digestibility over a range of kinetic characteristics. The steady-state solutions are particularly useful in understanding and predicting the depression in digestibility associated with changes in rates of passage at high levels of feed intake.

Hypothesis testing in analysis of variance requires that the errors be distributed normally and that subclass variances be homogeneous. The purpose of this study was to find the transformation for somatic cell concentration which meets these characteristics of hypothesis testing. Data consisted of 51,800 monthly tests from 52 herds on Dairy Herd Improvement in Wisconsin. Cell concentration by the Filter-DNA method was thousands of cells/ml. Analysis of untransformed data revealed extensive departure from normality and homogeneity. The family of transformations was Y' = (Y+M) L for L~0 or Y' = Log e (Y+M) for L=0, where Y was the untransformed cell concentration. The analysis included L ranging from -.7 to +.6 and M ranging from 0 to 20. The maximum likelihood estimate of L was zero. By log transformation, student's t for skewness and kurtosis and Chi square for heterogeneity of variance were not different from zero. Adding a constant of 10 before taking the log caused a small improvement in tests of normality and heterogeneity of variance.

Physical and chemical properties of cheese, such as texture, color, melt, and stretch, are primarily determined by the interaction of casein (CN) molecules. This review will discuss CN chemistry, how it is influenced by the cheese-making process, and how it impinges on the final product, cheese. We attempt to demonstrate that the application of principles governing the molecular interactions of CN can be useful in understanding the many physical and chemical properties of cheese and, in turn, how this can be used by the cheesemaker to produce the desired cheese. The physical properties of cheese (as well as flavor) are influenced by a number of factors including: milk composition; milk quality; temperature; the rate and extent of acidification by the starter bacteria; the pH history of cheese; the concentration of Ca salts (proportions of soluble and insoluble forms); extent and type of proteolysis, and other ripening reactions. Our hypothesis is that these factors also control and modify the nature and strength of CN interactions. The approach behind the recently proposed dual-binding model for the structure and stability of CN micelles is used as a framework to understand the physical and chemical properties of cheese.

Twenty-four multiparous dairy cows (eight with ruminal cannulae) were blocked by days in milk and assigned to six balanced 4 x 4 Latin squares with 21-d periods. The four diets, formulated from alfalfa silage plus a concentrate mix based on ground high moisture ear corn, contained (dry matter basis): 1) 20% concentrate, 80% alfalfa silage (24% nonfiber carbohydrate; NFC), 2) 35% concentrate, 65% alfalfa silage (30% NFC), 3) 50% concentrate, 50% alfalfa silage (37% NFC), or 4) 65% concentrate, 35% alfalfa silage (43% NFC). Soybean meal and urea were added to make diets isonitrogenous with equal nonprotein nitrogen (NPN) (43% of total N). Total urine was collected with indwelling Folley catheters for 24 h during each period. There was no effect of diet on urinary creatinine excretion (average 29 mg/kg of BW/d). There were quadratic effects of diet on total urinary ecretion of allantoin, uric acid, and purine derivatives (allantoin plus uric acid), and on ruminal synthesis of microbial N estimated from purine derivatives; maxima occurred at about 35% dietary NFC. Urinary excretion also was estimated with spot urine samples from creatinine concentration and the mean daily creatinine excretion. Daily excretion of allantoin, uric acid, and purine derivatives estimated from spot urine sampling followed the same pattern as that observed with total collection; differences between measured and estimated urine volume were significant only for 35% dietary concentrate. Spot urine sampling appeared to yield satisfactory estimates of purine derivative excretion. Maximal urea N excretion was estimated to occur at about 31% dietary NFC. Milk allantoin secretion increased linearly with concentrate and accounted for 4 to 6% of the total purine derivative excretion. Microbial yield was maximal at 35% dietary NFC, suggesting that this was the optimal level for utilization of dietary NPN from alfalfa silage and other sources.

Starch is a nutritionally important carbohydrate in feeds that is increasingly measured and used for formulation of animal diets. Discontinued production of the enzyme Rhozyme-S required for AOAC Method 920.40 invalidated this method for starch in animal feeds. The objective of this study was to compare methods for the determination of starch as potential candidates as a replacement method and for an AOAC collaborative study. Many starch methods are available, but they vary in accuracy, replicability, and ease of use. After assays were evaluated that differed in gelatinization method, number of reagents, and sample handling, and after assays with known methodological defects were excluded, 3 enzymatic-colorimetric assays were selected for comparison. The assays all used 2-stage, heat-stable, a-amylase and amyloglucosidase hydrolyses, but they differed in the gelatinization solution (heating in water, 3-(N-morpholino) propanesulfonic acid buffer, or acetate buffer). The measured values included both starch and maltooligosaccharides. The acetate buffer-only method was performed in sealable vessels with dilution by weight; it gave greater starch values (2-6 percentage units of sample dry matter) in the analysis of feed/food substrates than did the other methods. This method is a viable candidate for a collaborative study.

column 1, line 14: "fuels and materials" should read "organic fuels and materials."Page 529, Table 4, line 10: "smithiti" should read "smithii."Page 530, column 2, line 5 from bottom: reference 651 should be deleted.Page 531, Table 5, Binding affinity: for CenA BMCC, "41" and "1.89" should read "8.6" and "0.40," respectively; for Cex BMCC, "33" and "1.71" should read "6.44" and "0.33," respectively.Page 537, line 10 from bottom: "n/(n Ϫ 1)" should read "(n Ϫ1)/n."Page 542, column 2, line 19 from bottom, "scale of availability feedstock" should read "scale of feedstock availability."Page 546, column 2, line 10: "involves naturally occurring cellulolytic microorganisms" should read "involves engineering naturally occurring cellulolytic microorganisms."Page 547, column 2, line 5 from bottom: "cellulolytic" should read "noncellulolytic."