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Single-pass corn stover harvest is a method whereby combine harvester tailings consisting of cob, stalk, leaf, husk, and tassel fractions are collected and baled without coming into contact with soil. The objective was to feed beef steers diets that included a roughage component consisting of harvested corn residue in chopped form from conventional corn stover bales (CST) or single-pass bales (SPB) to assess intake selectivity of corn stover fractions and estimate net energy values of these corn stovers. Whole plant corn silage served as the control roughage in the control diet (CSIL). Steers (n = 90, 5 pens per treatment) were fed during Grow (84 d) and Finish (66 d) trials. Steers sorted through corn stover during both trials and consumed 52.5% of corn stover offered and 40% of cob offered. Intake of SPB cob was 2.6-fold and 3.3-fold greater than CST cob intake (P < 0.01) in Grow and Finish trials, respectively, indicating that when more cob was available, cattle consumed more. During the Grow trial, stover intake tended (P = 0.07) to be lower for SPB and diet dry matter intake (DMI) was less (P ≤ 0.05) for SPB, which may be due to the elevated cob intake by SPB steers. Across treatments, stover DMI was similar in the Grow (1.10 kg per steer per d) and Finish (1.11 kg per steer per d) trials. The proportion (65%) of Fines (

Three alfalfa biotypes were chosen based on the presumption that they would be sources of alfalfa herbage that differed in lignin concentration and therefore cell wall digestibility. The hypothesis was that a lesser lignin concentration would result in greater alfalfa neutral detergent fiber (NDF) digestibility and greater beef steer growth performance. The three alfalfa biotypes were HarvXtra (Forage Genetics International), Hi-Gest 360 (Alforex Seeds), and a control alfalfa, LegenDairy XHD (Winfield Solutions LLC). High-moisture wrapped bales were prepared from second-harvest, d 30 crops. Digestibility of NDF was determined using in vitro incubations and a steer digestibility trial. Alfalfa baleage and trace mineral salt were fed to Angus steers (300 kg initial body weight, 4 pens/treatment) in an 83-day growing-phase trial. Alfalfa acid detergent lignin concentrations were 75.6, 71.8, and 63.0 g/kg dry matter (P = 0.34) for LegenDairy, Hi-Gest and HarvXtra, respectively. Based on in vitro total-tract NDF digestibility coefficients, HarvXtra tended (P ≥ 0.09) to have the highest NDF digestibility. Alfalfa biotype affected in vivo apparent total tract digestibility of NDF (P < 0.001) and there was a trend for an effect on acid detergent fiber digestibility (P = 0.051). Hi-Gest and HarvXtra had similar in vivo apparent NDF digestibilities, which were greater than for LegenDairy (P < 0.05). There was no alfalfa biotype effect on daily alfalfa dry matter intake (DMI; P = 0.51) or average daily gain (P = 0.25) by growing steers. The absence of an effect by the novel alfalfa biotypes on DMI by growing steers suggests that the compositional and digestibility differences of the novel alfalfa biotypes compared to LegenDairy were not sufficient to alleviate the limitation of physical fill (if evident) on DMI. If more disparity in cell wall composition and NDF digestibility were to exist between control and reduced-lignin biotypes, then perhaps an advantage in cattle growth performance for a reduced-lignin alfalfa biotype would be detectable.

Although more than 3 million head of dairy cows enter the food supply chain in the U.S. every year, research on this topic remains limited and scarce. Meat production from dairy cows is a significant component of beef production, accounting for almost 10% of U.S. commercial beef production. Thus, the purpose of this review is to demonstrate the importance of dairy cows as a beef source, and to provide an overview on topics from farm to meat product—culling, marketing, transportation, welfare, body composition and its relationship with lactation particularities, carcass characteristics, meat quality, and traceability. Current scientific evidence has shown that culling a dairy cow at an appropriate time has beneficial effects on cow welfare and, consequently, cow value. During marketing the dairy cow is visually evaluated for health and factors associated with its expected carcass value; thus, marketing a well-conditioned cow will ensure that the animal is fit for transportation and provides high carcass yield. However, limitations such as low body condition score, lameness or mobility problems, and visual defects remain persistent. Even though beef harvest plants accommodate cows in all body composition states, the current carcass grade system does not reflect the mature cow industry needs. Therefore, improvement of the grading system could maximize carcass utilization and increase cow carcass value by recognizing subprimal cuts that could be merchandised as whole muscle cuts. Lastly, implementation of a traceability system would unify information from the farm to harvest assisting the industry in making further advancements.

Cull dairy cows contribute almost 10% of national beef production in the United States. However, different factors throughout the life of dairy cows affect their weight and overall body condition as well as carcass traits, and consequently affect their market price. Therefore, the objectives of this study were: (1) to assess relationships between price ratio and carcass merit of cull dairy cows sold through several sites of an auction market and (2) to investigate the effect of animal life history events and live weight on sale barn price (BP) and price ratio (as a measure of relative price), as an indicator of carcass merit. Data from 4 dairy operations included 3,602 cull dairy cow records during the period of 2015 to 2019. Life history events data were collected from each dairy operation through Dairy Comp software; live weight and price were obtained periodically from the auction market, and the carcass data were provided by a local packing plant. Cow price in dollars per unit of live weight ($/cwt) and price ratio were the 2 outcome variables used in the analyses. Price ratio was created aiming to remove seasonality effects from BP (BP divided by the national average price for its respective month and year of sale). The association between price ratio and carcass merit traits was investigated using canonical correlation analysis, and the effect of life history events on both BP and price ratio was inferred using a multiple linear regression technique. More than 70% of the cows were culled in the first 3 lactations, with an average live weight of 701.5 kg, carcass weight of 325 kg, and dressing percentage of 46.3%. On average, cull cows were sold at $57.0/cwt during the period considered. The canonical correlation between price ratio and carcass merit traits was 0.76, indicating that price ratio reflected carcass merit of cull cows. Later lactations led to lower BP compared with cows culled during the first 2 lactations. Injury, and leg and feet problems negatively affected BP. Productive variables demonstrated that the greater milk production might lead to lower cow prices. A large variation between farms was also noted. In conclusion, price ratio was a good indicator of carcass merit of cull cows, and life history events significantly affected sale BP and carcass merit of cull cows sold through auction markets.

Clinical signs of photosensitivity in cattle can occur sporadically and unpredictably. It is believed that cases of photosensitivity may be underreported, causing inaccurate and inflated reports of mortality. Additionally, because secondary photosensitization in grazing cattle occurs with liver damage or dysfunction, photosensitivity can have many potential or associated causes. This case links a previous occurrence of coccidiosis to an outbreak of photosensitivity in grazing Holstein steers. Grazing management staff first observed clinical signs of photosensitivity 17 days after an outbreak of coccidiosis and subsequent turnout to spring pastures. Clinical signs were observed in 25% of the population. The severity of photosensitivity was variable and ranged from blistered skin on the muzzle to sloughing of unpigmented epidermis and thinly haired regions. Severely affected cattle were removed from pasture, housed under shade, monitored for infection, and recovered without treatment. Mild cases remained on pasture and recovered without treatment. Photosensitivity did not reoccur in the cattle that remained on pasture or in mildly affected cattle returned to pasture. Photosensitivity did not appear to be associated with pasture weeds, a specific forage species, or variable or extreme weather conditions that could have resulted in mycotoxin production. The occurrence appears to have been a result of a previous and concurrent coccidiosis outbreak that caused secondary photosensitization through hepatic lipidosis caused by anorexia and dehydration associated with the severe coccidiosis. Although clinical signs appeared suddenly, cattle recovered quickly and without treatment.

Three different methods of wrapping and storing alfalfa hay in round bales were used to explore possible differences in preferential consumption. Large round bales of alfalfa were stored indoors, outdoors with conventional net wrap, or outdoors with a new wrap that incorporates a breathable film. This film is intended to better conserve hay in round bales by shedding precipitation but allowing internal moisture to exit the bale through microscopic pores. Five separate preference trials, each of 18-d duration with six 3-d periods, were conducted using beef cattle. In all 5 preference trials, hay wrapped with breathable film was preferred over net-wrapped hay stored outdoors. For instance, in trial 1 hay wrapped with breathable film was preferred over net-wrapped hay stored outdoors in all 18 pairings (P < 0.001), and consumption of the film-wrapped hay was significantly greater (78 vs. 28% of total hay offered, P = 0.05). Results suggest that when bales are stored outdoors, cattle will strongly prefer to consume hay from bales wrapped with breathable film compared with net-wrapped bales. Although in 2 trials hay wrapped with breathable film was preferred over hay stored indoors, when considered across all trials and pairings, preference of hay from breathable film bales did not differ from that stored indoors.

The effect of dietary nitrate supplementation on rumen bacterial community composition was examined in beef steers fed either a nitrate-N diet or urea-N diet. An automated method of ribosomal intergenic spacer analysis was applied to solid and liquid fractions of ruminal contents to allow comparison of bacterial communities. Supplemental N source affected relative population size of four amplicon lengths (ALs) in the liquid fraction and three ALs in the solid fraction. Five ALs were more prevalent after adaptation to nitrate. Correspondence analysis indicated that feeding the steers the nitrate-N diet versus urea-N diet changed the bacterial community composition in the liquid but not in the solid fraction. This led to an investigation of the relative sizes of potential nitrate-reducing populations. Mannheimia succiniciproducens, Veillonella parvula, and Campylobacter fetus were obtained from nitrate enrichment culture and quantified by real-time PCR based on 16S rRNA sequence. Nitrate supplementation increased the percentage of C. fetus in the liquid and solid phases, and in solid phase, the percentage of M. succiniciproducens increased. No change in species prevalence was observed for V. parvula. However, even after adaptation to dietary nitrate, the relative population sizes for all three putative nitrate-reducing species were very low (

The objectives were to compare the ability of various rumen microbial fractions to reduce nitrate and to assess the effect of nitrate on in vitro fermentation characteristics. Physical and chemical methods were used to differentiate the rumen microbial population into the following fractions: whole rumen fluid (WRF), protozoa (Pr), bacteria (Ba), and fungi (Fu). The three nitrogen substrate treatments were as follows: no supplemental nitrogen source, nitrate or urea, with the latter two being isonitrogenous additions. The results showed that during 24 h incubation, WRF, Pr and Ba fractions had an ability to reduce nitrate, and the rate of nitrate disappearance for the Pr fraction was similar to the WRF fraction, while the Ba fraction needed an adaptation period of 12 h before rapid nitrate disappearance. The WRF fraction had the greatest methane () production and the Pr fraction had the greatest prevailing concentration (p production during incubation (p-N) concentration (p production by all microbial fractions.

A study of Escherichia coli O157:H7 transmission and shedding was conducted with bull calves housed in individual pens within a confined environment. For comparative purposes, the numbers and duration of E. coli O157:H7 shedding in naturally infected calves were monitored after a single purchased calf (calf 156) tested positive prior to inoculation. During the next 8 days, the calves in adjacent pens and a pen directly across a walkway from calf 156 began to shed this serotype O157:H7 strain. Five of the eight calves in this room shed this O157:H7 strain at some time during the following 8 weeks. The numbers of E. coli O157:H7 isolates shed in these calves varied from 60 to 10 5 CFU/g of feces, and the duration of shedding ranged from 17 to >31 days. The genomic DNAs from isolates recovered from these calves were indistinguishable when compared by using Xba I digestion and pulsed-field gel electrophoresis. Inoculation of calves with 1 liter of water containing ca. 10 3 to 10 4 CFU of E. coli O157:H7/ml resulted in shedding in 10 of 12 calves (trial 1, 4 of 4 calves; trial 2, 6 of 8 calves). The inoculated calves shed the inoculation strain (FRIK 1275) as early as 24 h after administration. The duration of shedding varied from 18 to >43 days at levels from 10 2 to 10 6 CFU/g of feces. The numbers of doses necessary to initiate shedding varied among calves, and two calves in trial 2 never shed FRIK 1275 after four doses (ca. 10 6 CFU per dose). Results from this study confirm previous reports of animal-to-animal and waterborne dissemination of E. coli O157:H7 and highlight the need for an effective water treatment to reduce the spread of this pathogen in cattle.

Two experiments were conducted to evaluate feedlot performance of heifers and calves and the lactational characteristics and carcass composition and quality of heifers while in the single-calf heifer (SCH) system. In Exp. 1, 24 lactating Angus × Holstein heifers were slowly adapted to an 85% concentrate diet, and one-half of the heifers were implanted with Finaplix-H®, both at 11 wk postpartum. Heifer-calf pairs were then placed in feedlot pens, equipped with pinpointer feeding devices, and fed an 85% concentrate diet for 102 d. The control (C) and implanted (I) heifers had similar (P=0.27) daily gains and similar (P=0.30) DMI; however, I heifers consumed 8.9% less (P

Two experiments were conducted to evaluate feedlot performance, lactational characteristics, and carcass composition and quality of heifers and the performance of their calves in a single-calf heifer (SCH) system. In Exp. 1, 13 [10 lactating (L) and 3 nonlactating (NL)] prenatally androgenized (PA) heifers, born to cows implanted with testosterone propionate (TP) and 19 (13 L and 6 NL) control (C) heifers, born to nonimplanted cows, were used. Heifers were calved and the pairs were placed in feedlot pens to evaluate the effects of PA on feedlot performance and lactation. Heifers were fed an 85% concentrate diet and fed to a compositional endpoint of 1.1 cm of subcutaneous fat cover, at which point calves were weaned and heifers slaughtered approximately 12 h later. The NL heifers consumed 17.0% less (P

The objective of this study was to assess fermentation product, growth rate and growth yield responses of Selenomonas ruminantium HD4 to limiting and non-limiting ammonia concentrations. The ammonia half-inhibition constant for S. ruminantium in batch culture was 296 mM. Cells were grown in continuous culture with a defined ascorbate-reduced basal medium containing either 0.5, 5, 25, 50, 100 or 200 mM NH4Cl and dilution rates were 0.07, 0.14, 0.24 or 0.40 h-1. Ammonia was the growth-limiting nutrient when 0.5 mM NH4Cl was provided and the half-saturation constant was 72 microM. Specific rates of glucose utilization and fermentation acid carbon formation were highest for 0.5 mM NH4Cl. Lactate production (moles per mole of glucose disappearing) increased at the fastest dilution rate (0.40 h-1) for 5.0 mM NH4Cl while acetate and propionate decreased when compared to slower dilutions (0.07 and 0.14 h-1). Lactate production remained low while acetate and propionate remained high for all dilution rates when NH4Cl concentrations were 25 mM or greater. Yield (Y(Glc) and Y(ATP)) were nearly doubled when NH4Cl was increased from 0.5 mM (25.1 g cells/mol glucose used and 13.9 g cells/mol ATP produced respectively) to the higher concentrations. Y(Glc) was highest at 25 mM and 50 mM NH4Cl (48.2 cells/mol and 43.1 cells/mol respectively) as was Y(ATP) (23.2 cells/mol and 20.8 cells/mol respectively). Y(NH3) was highest at the lowest NH4Cl concentration. The maximal fermentation product formation rate occurred at a growth-limiting ammonia concentration, while maximal glucose and ATP bacterial yields occurred at non-growth-limiting ammonia concentrations. Given the growth response of this ruminal bacterium, it is possible that maximization of ruminal bacterial yield may necessitate sacrificing the substrate degradation rate and vice versa.

A study was conducted to evaluate the main effects of dietary nitrate adaptation by cattle and alfalfa cell wall to starch ratio in in vitro substrates on nitrate disappearance and nitrite and volatile fatty acid (VFA) concentrations, as well as hydrogen (H2) and methane (CH4) accumulations. Rumen fluid from steers fed diets containing urea or nitrate was added into in vitro incubations containing sodium nitrate as the sole nitrogen source and 20 cell wall : 80 starch or 80 cell wall : 20 starch as the carbohydrate source. The results showed that during 24 h incubation, rumen fluid inoculums from steers adapted to dietary nitrate resulted in more rapid nitrate disappearance by 6 h of incubation (P < 0.01), no significant effect on nitrite concentration and diminished CH4 accumulation (P < 0.05). Cell wall to starch ratio did not affect nitrate disappearance, CH4 accumulation and total VFA concentration. The higher cell wall ratio had the lower total gas production and H2 concentration (P < 0.05). Ammonia-N (NH3-N) concentration increased because of adaptation of donors to nitrate feeding (P < 0.05). Nitrate adaptation did not alter total VFA concentration, but increased acetate, and decreased propionate and butyrate molar proportions (P < 0.01).

The specificity of polyclonal antibodies prepared against strains of Selenomonas ruminantium, the effect of assay conditions, and quantification of individual strains in mixed-cell suspensions of selenomonad strains were examined in this study. Whole-cell suspensions were prepared with pure cultures of S. ruminantium PC18, HD 4 , GA192, and D. Each cell suspension was injected into a Leghorn laying hen, and polyclonal antibodies were harvested from eggs laid in week 3 or 7 following initial immunization. Antibodies made to the S. ruminantium strains readily discerned the homologous strain from the heterologous strains. Cross-reactivity among antibodies and the heterologous S. ruminantium strains ranged from 5 to 26%. Among non- S. ruminantium species, cross-reactivity of S. ruminantium antibodies was greatest with Selenomonas sputigena (3 to 34%) and Succinivibrio dextrinosolvens (0 to 37%). Antibodies made to strains GA192 and D were used to quantify a mixture of the two strains. Both antibodies responded to graded concentrations of the homologous antigen in the biculture mixtures in accord with the change in the direct cell counts for each strain (strain D, R 2 = 0.92; strain GA192, R 2 = 0.90). This enzyme-linked immunosorbent assay enabled concurrent and accurate quantification of two strains of S. ruminantium subsp. ruminantium in a mixed-cell suspension with a precision of much less than 1 order of magnitude.

The objective of this study was to evaluate the role of reductive acetogenesis as an alternative H 2 disposal mechanism in the rumen. H 2 /CO 2 -supported acetogenic ruminal bacteria were enumerated by using a selective inhibitor of methanogenesis, 2-bromoethanesulfonic acid (BES). Acetogenic bacteria ranged in density from 2.5 × 10 5 cells/ml in beef cows fed a high-forage diet to 75 cells/ml in finishing steers fed a high-grain diet. Negligible endogenous acetogenic activity was demonstrated in incubations containing ruminal contents, NaH 13 CO 3 , and 100% H 2 gas phase since [U- 13 C]acetate, as measured by mass spectroscopy, did not accumulate. Enhancement of acetogenesis was observed in these incubations when methanogenesis was inhibited by BES and/or by the addition of an axenic culture of the rumen acetogen Acetitomaculum ruminis 190A4 (10 7 CFU/ml). To assess the relative importance of population density and/or H 2 concentration for reductive acetogenesis in ruminal contents, incubations as described above were performed under a 100% N 2 gas phase. Both selective inhibition of methanogenesis and A. ruminis 190A4 fortification (>10 5 CFU/ml) were necessary for the detection of reductive acetogenesis under H 2 -limiting conditions. Under these conditions, H 2 accumulated to 4,800 ppm. In contrast, H 2 accumulated to 400 ppm in incubations with active methanogenesis (without BES). These H 2 concentrations correlated well with the pure culture H 2 threshold concentrations determined for A. ruminis 190A4 (3,830 ppm) and the ruminal methanogen 10-16B (126 ppm). The data demonstrate that ruminal methanogenic bacteria limited reductive acetogenesis by lowering the H 2 partial pressure below the level necessary for H 2 utilization by A. ruminis 190A4.

Background Chronic, low-grade inflammation provides a link between normal ageing and the pathogenesis of age-related diseases. A series of in vitro tests confirmed the strong anti-inflammatory activities of known inhibitors of NF-κB activation (δ-tocotrienol, quercetin, riboflavin, (-) Corey lactone, amiloride, and dexamethasone). δ-Tocotrienol also suppresses β-hydroxy-β-methylglutaryl coenzyme A (HMG-CoA) reductase activity (the rate-limiting step in de novo cholesterol synthesis), and concomitantly lowers serum total and LDL cholesterol levels. We evaluated these compounds in an avian model anticipating that a dietary additive combining δ-tocotrienol with quercetin, riboflavin, (-) Corey lactone, amiloride, or dexamethasone would yield greater reductions in serum levels of total cholesterol, LDL-cholesterol and inflammatory markers (tumor necrosis factor-α [TNF-α], and nitric oxide [NO]), than that attained with the individual compounds. Results The present results showed that supplementation of control diets with all compounds tested except riboflavin, (-) Corey lactone, and dexamethasone produced small but significant reductions in body weight gains as compared to control. (-) Corey lactone and riboflavin did not significantly impact body weight gains. Dexamethasone significantly and markedly reduced weight gain (>75%) compared to control. The serum levels of TNF-α and NO were decreased 61% - 84% (P < 0.001), and 14% - 67%, respectively, in chickens fed diets supplemented with δ-tocotrienol, quercetin, riboflavin, (-) Corey lactone, amiloride, or dexamethasone as compared to controls. Significant decreases in the levels of serum total and LDL-cholesterol were attained with δ-tocotrienol, quercetin, riboflavin and (-) Corey lactone (13% - 57%; P < 0.05), whereas, these levels were 2-fold higher in dexamethasone treated chickens as compared to controls. Parallel responses on hepatic lipid infiltration were confirmed by histological analyses. Treatments combining δ-tocotrienol with the other compounds yielded values that were lower than individual values attained with either δ-tocotrienol or the second compound. Exceptions were the significantly lower total and LDL cholesterol and triglyceride values attained with the δ-tocotrienol/(-) Corey lactone treatment and the significantly lower triglyceride value attained with the δ-tocotrienol/riboflavin treatment. δ-Tocotrienol attenuated the lipid-elevating impact of dexamethasone and potentiated the triglyceride lowering impact of riboflavin. Microarray analyses of liver samples identified 62 genes whose expressions were either up-regulated or down-regulated by all compounds suggesting common impact on serum TNF-α and NO levels. The microarray analyses further identified 41 genes whose expression was differentially impacted by the compounds shown to lower serum lipid levels and dexamethasone, associated with markedly elevated serum lipids. Conclusions This is the first report describing the anti-inflammatory effects of δ-tocotrienol, quercetin, riboflavin, (-) Corey lactone, amiloride, and dexamethasone on serum TNF-δ and NO levels. Serum TNF-δ levels were decreased by >60% by each of the experimental compounds. Additionally, all the treatments except with dexamethasone, resulted in lower serum total cholesterol, LDL-cholesterol and triglyceride levels. The impact of above mentioned compounds on the factors evaluated herein was increased when combined with δ-tocotrienol.

The objective of this study was to determine the maximum ammonium source concentration tolerated by Selenomonas ruminantium and its metabolic response to high ammonium source concentrations. The ammonia-nitrogen half-inhibition constant (K i) in defined basal medium was 239 mabetm for NH4Cl, 173 mabetm for NH4OH, 153 mabetm for (NH4)2SO4 and 110 mabetm for NH4HCO3. Reduction in continuous culture maximal growth rate was similar to the reduction in the batch culture logarithmic growth rate for the respective NH4Cl concentrations. Cell yield when expressed as Y ATP decreased for 150 and 200 mabetm NH4C1. the NH3-N K i estimates are in line with inhibiting concentrations observed for other bacteria and suggest that energy efficiency is reduced when the NH3-N concentration is increased.

Population development of key groups of bacteria involved in municipal refuse conversion to methane was measured from the time of initial incubation through the onset of methane production. Hemicellulolytic bacteria, cellulolytic bacteria, hydrogen-producing acetogens, and acetate- and H 2 -plus-CO 2 -utilizing methanogens were enumerated by the most-probable-number technique with media containing oat spelt xylan, ball-milled cellulose, butyrate, acetate, and H 2 plus CO 2 , respectively. Refuse decomposition was monitored in multiple replicate laboratory-scale sanitary landfills. A laboratory-scale landfill was dismantled weekly for microbial and chemical analysis. Leachate was neutralized and recycled to ensure methanogenesis. The methane concentration of the sampled containers increased to 64% by day 69, at which time the maximum methane production rate, 929 liters of CH 4 per dry kg-year, was measured. Population increases of 2, 4, 5, 5, and 6 orders of magnitude were measured between fresh refuse and the methane production phase for the hemicellulolytic bacteria, cellulolytic bacteria, butyrate-catabolizing acetogens, and acetate- and H 2 -CO 2 -utilizing methanogens, respectively. The cellulolytic bacteria and acetogens increased more slowly than the methanogens and only after the onset of methane production. The initial decrease in the pH of the refuse ecosystem from 7.5 to 5.7 was attributed to the accumulation of acidic end products of sugar fermentation, to the low acid-consuming activity of the acetogenic and methanogenic bacteria, and to levels of oxygen and nitrate in the fresh refuse sufficient for oxidation of only 8% of the sugars to carbon dioxide and water. Cellulose and hemicellulose decomposition was most rapid after establishment of the methanogenic and acetogenic populations and a reduction in the initial accumulation of carboxylic acids. A total of 72% of these carbohydrates were degraded in the container sampled after 111 days. Initially acetate utilization, but ultimately polymer hydrolysis, limited the rate of refuse conversion to methane. Microbial and chemical composition data were combined to formulate an updated description of refuse decomposition in four phases: an aerobic phase, an anaerobic acid phase, an accelerated methane production phase, and a decelerated methane production phase.

Techniques were evaluated for formation of a liquid inoculum from shredded municipal refuse, including chilling the refuse at 4°C prior to blending and multiple washing and blending cycles. The average count of cellulolytic bacteria from six different detachment treatments was 5.1 × 10 4 cells per g (dry weight) of refuse with a range of 0.7 × 10 4 to 12.7 × 10 4 cells per g (dry weight). The liquid obtained from blending the refuse in phosphate buffer followed by hand squeezing was the selected detachment procedure. The inoculum formation procedure was validated by the addition of ruminal cellulolytic bacteria to refuse and recovery of the cellulolytic bacteria by most-probable-number enumerations. The ratio of measured to expected cell counts among tests in which different volumes of ruminal fluid were added to refuse ranged from 2.7 to 14.4. There was no evidence of anaerobic cellulolytic fungi in a refuse sample.