Comparative in vitro evaluation of forage legumes (prosopis, acacia, atriplex, and leucaena) on ruminal fermentation and methanogenesis

Two experiments in vitro were conducted to evaluate four Egyptian forage legume browses, i.e., leaves of prosopis (Prosopis juliflora), acacia (Acacia saligna), atriplex (A triplex halimus), and leucaena (Leucaena leucocephala), in comparison with Tifton (Cynodon sp.) grass hay for their gas production, methanogenic potential, and ruminal fermentation using a semi-automatic system for gas production (first experiment) and for ruminal and post ruminal protein degradability (second experiment). Acacia and leucaena showed pronounced methane inhibition compared with Tifton, while prosopis and leucaena decreased the acetate:propionate ratio (P<0.01). Acacia and leucaena presented a lower (P<0.01) ruminal NH3-N concentration associated with the decreasing (P<0.01) ruminal protein degradability. Leucaena, however, showed higher (P<0.01) intestinal protein digestibility than acacia. This study suggests that the potential methanogenic properties of leguminous browses may be related not only to tannin content, but also to other factors.

Predicting Growth of Panicum maximum: An Adaptation of the CROPGRO-Perennial Forage Model

Warm-season grasses are economically important for cattle production in tropical regions and tools to aid in management and research on these forages would be highly beneficial both in research and the industry. This research was conducted to adapt the CROPGRO-Perennial Forage model to simulate growth of the tropical species guineagrass (Panicum maximum Jacq. cv. 'Tanzania') and to describe model adaptation for this species. To develop the CROPGRO parameters for this species, we began with values and relationships reported in the literature. Some parameters and relationships were calibrated by comparison with observed growth, development, dry matter accumulation, and partitioning during a 17-mo experiment with Tanzania guineagrass in Piracicaba, SP, Brazil. Compared with starting parameters for palisadegrass [Brachiaria brizantha (A. Rich.) Stapf. cv. 'Xaraes'], dormancy effects of the perennial forage model had to be minimized, partitioning to storage tissue or root decreased, and partitioning to leaf and stem increased to provide for more leaf and stem growth and less root. Parameters affecting specific leaf area and senescence of plant tissues were improved. After these changes were made to the model, biomass accumulation was better simulated, mean predicted herbage yield was 6576 kg ha(-1), averaged across 11 regrowth cycles of 35 (summer) or 63 d (winter), with a RMSE of 494 kg ha(-1) (Willmott's index of agreement d = 0.985, simulated/observed ratio = 1.014). The model also gave good predictions against an independent data set, with similar RMSE, ratio, and d. The results of the adaptation suggest that the CROPGRO model is an efficient tool to integrate physiological aspects of guineagrass and can be used to simulate growth.

Chemical characterization and in vitro biological activity of four tropical legumes, Styzolobium aterrimum L., Styzolobium deeringianum, Leucaena leucocephala, and Mimosa caesalpiniaefolia, as compared with a tropical grass, Cynodon spp. for the use in ruminant diets

Leucaena leucocephala (LEU) and three under-utilized tanniferous legumes, Styzolobium aterrimum L. (STA), Styzolobium deeringianum (STD), and Mimosa caesalpiniaefolia Benth (MIC) were chemically characterized and the biological activity of tannins was evaluated using in vitro simulated ruminal fermentation through tannin-binding polyethylene glycol (PEG) and compared with a non-tanniferous tropical grass hay, Cynodon spp. (CYN). The Hohenheim gas test was used and gas production (GP) was recorded at 4, 8, 12, 24, 32, 48, 56, 72, 80, and 96 h incubation with and without PEG. Kinetic parameters were estimated by an exponential model. STA, STD, and LEU contained higher (P < 0.05) crude protein than MIC, which had greater neutral detergent fibre and acid detergent fibre. Total phenols, total tannins, and condensed tannins (CT) were consistently the highest in MIC. Gas production was the lowest from MIC (P < 0.05) and the highest in LEU and STA. MIC + PEG largely reduced (P < 0.05) the lag phase and the fractional rate of fermentation and increased potential GP. Kinetic parameters of STA + PEG and LEU + PEG were not affected. LEU + PEG produced greater gas increment (P < 0.05) than STD + PEG, although both legumes had the same CT. All legumes except MIC were more extensively degraded than CYN. However, fermentation of the legumes was differently affected by the presence and proportions of CT, indigestible fibre or both.

Preference of dairy cows for ryegrass, white clover and red clover, and its effects on nutrient supply and milk quality

Two experiments were conducted with 30 dairy cows each, to study the preference for fresh (Experiment 1) and ensiled (Experiment 2) ryegrass, white and red clover. Both experiments consisted of three choice diets with white or red clover or both, offered with ryegrass, and two diets with ryegrass mixed with white or red clover (40% clover). Cows consumed diets with 37.7% fresh white and 45.9% red clover, and no preference was observed when the cows were offered all three forages. By contrast, cows preferred white and red clover silage (73.0 and 69.2%, respectively) over ryegrass silage (of lower nutritive quality). When offered three forages, cows preferred white (59.8%) over red clover (17.5%) and ryegrass (22.7%). Choice diets resulted in diets similar (fresh forages) or higher in nutrient content and digestibility (silages). Treatments did not affect feed intake and performance. Choices compared to mixed diets with red clover silage were preferable regarding the fatty acid composition of the milk fat. Obviously, only large differences in nutrient and energy concentration facilitate preferences for clovers over ryegrass, which could, depending on clover type, be beneficial in terms of the milk's fatty acid composition.

Intensive Rotational Grazing of Steers on Highly Erodible Land at the Adams County CRP Project

Grazing yearling steers is one way to utilize the forages required for participation in the Conservation Reserve Program (CRP) after CRP contracts expire. In 1995, a stocker-steer intensive-rotational grazing study was conducted at the CRP Research and Demonstration Project near Corning, Iowa. A similar study was carried out in 1994. Seventy-five yearling crossbred steers grazed a 65- acre pasture that had been divided into 27 paddocks using electric fencing from May 4, 1995 to September 14, 1995. During this period, the 65-acre pasture system produced 9,975 animal-days of grazing and 11,403 pounds of gain. On a per-acre basis, this translates to 153.5 animal-days of grazing and 175.4 pounds of gain. The stocking rate was constant for the entire 133- day grazing season at 1.15 steers per acre. On May 4, 1995, the beginning of the grazing season, the average weight of the steers was 495.7 pounds. By the end of the grazing trial on September 14, 1995, the average weight of the steers had increased to 647.7 pounds. The average gain per steer during the 133-day grazing period was 152 pounds, and the average daily gain per steer was 1.14 pounds. The average bodyweight of the steers during the entire grazing season was 571.7 pounds.

Integrating the Use of Spring- and Fall-Calving Beef Cows in a Year-round Grazing System (A Progress Report)

Animal production, hay production and feeding, and the yields and composition of forage from summer and winter grass-legume pastures and winter corn crop residue fields from a year-round grazing system were compared with those of a conventional system. The year-round grazing system utilized 1.67 acres of smooth bromegrass-orchardgrass-birdsfoot trefoil pasture per cow in the summer, and 1.25 acres of stockpiled tall fescue-red clover pasture per cow, 1.25 acres of stockpiled smooth bromegrass-red clover pasture per cow, and 1.25 acres of corn crop residues per cow during winter for spring- and fall-calving cows and stockers. First-cutting hay was harvested from the tall fescue-red clover and smooth bromegrass-red clover pastures to meet supplemental needs of cows and calves during winter. In the conventional system (called the minimal land system), spring-calving cows grazed smooth bromegrass-orchardgrass-birdsfoot trefoil pastures at 3.33 acres/cow during summer with first cutting hay removed from one-half of these acres. This hay was fed to these cows in a drylot during winter. All summer grazing was done by rotational stocking for both systems, and winter grazing of the corn crop residues and stockpiled forages for pregnant spring-calving cows and lactating fall-calving cows in the year-round system was managed by strip-stocking. Hay was fed to springcalving cows in both systems to maintain a mean body condition score of 5 on a 9-point scale, but was fed to fall-calving cows to maintain a mean body condition score of greater than 3. Over winter, fall-calving cows lost more body weight and condition than spring calving cows, but there were no differences in body weight or condition score change between spring-calving cows in either system. Fall- and spring-calving cows in the yearround grazing system required 934 and 1,395 lb. hay dry matter/cow for maintenance during the winter whereas spring-calving cows in drylot required 4,776 lb. hay dry matter/cow. Rebreeding rates were not affected by management system. Average daily gains of spring-born calves did not differ between systems, but were greater than fall calves. Because of differences in land areas for the two systems, weight production of calves per acre of cows in the minimal land system was greater than those of the year-round grazing system, but when the additional weight gains of the stocker cattle were considered, production of total growing animals did not differ between the two systems.

Evaluation of Year-round Forage Management Systems for Spring- and Fall-Calving Beef Cows (A Progress Report)

A year-round grazing system for spring- and fall-calving cows was developed to compare animal production and performance, hay production and feeding, winter forage composition changes, and summer pasture yield and nutrient composition to that from a conventional, or minimal land system. Systems compared forage from smooth bromegrass-orchardgrass-birdsfoot trefoil pastures for both systems in the summer and corn crop residues and stockpiled grass-legume pastures for the year-round system to drylot hay feeding during winter for the minimal land system. The year-round grazing system utilized 1.67 acres of smooth bromegrassorchardgrass- birdsfoot trefoil (SB-O-T) pasture per cow in the summer, compared with 3.33 acres of (SB-O-T) pasture per cow in the control (minimal land) system. In addition to SB-O-T pastures, the year-round grazing system utilized 2.5 acres of tall fescue-red clover (TFRC) and 2.5 acres of smooth bromegrass-red clover (SBRC) per cow for grazing in both mid-summer and winter for fall- and spring-calving cows, respectively. First-cutting hay was harvested from the TF-RC and SB-RC pastures, and regrowth was grazed for approximately 45 days in the summer. These pastures were then fertilized with 40 lbs N/acre and stockpiled for winter grazing. Also utilized during the winter for spring-calving cows in the year-round grazing system were corn crop residue (CCR) pastures at an allowance of 2.5 acres per cow. In the minimal land system, hay was harvested from three-fourths of the area in SB-O-T pastures and stored for feeding in a drylot through the winter. Summer grazing was managed with rotational stocking for both systems, and winter grazing of stockpiled forages and corn crop residues by year-round system cows was managed by strip-stocking. Hay was fed to maintain a body condition score of 5 on a 9 point scale for spring-calving cows in both systems. Hay was supplemented as needed to maintain a body condition score of 3 for fall-calving cows nursing calves through the winter. Although initial condition scores for cows in both systems were different at the initiation of grazing for both winter and summer, there were no significant differences (P > .05) in overall condition score changes throughout both grazing seasons. In year 1, fall-calving cows in the year-round grazing system lost more (P < .05) body weight during winter than spring-calving cows in either system. In year 2, there were no differences seen in weight changes over winter for any group of cows. Average daily gains of fall calves in the yearround system were 1.9 lbs/day compared with weight gains of 2.5 lbs/day for spring calves from both systems. Yearly growing animal production from pastures for both years did not differ between systems when weight gains of stockers that grazed summer pastures in the year-round grazing system were added to weight gains of suckling calves. Carcass characteristics for all calves finished in the feedlot for both systems were similar. There were no significant differences in hay production between systems for year 1; however, amounts of hay needed to maintain cows were 923, 1373, 4732 lbs dry matter/cow for year-round fall-calving, year-round spring-calving, and minimal land spring-calving cows, respectively. In year 2, hay production per acre in the minimal land system was greater (P < .05) than for the year-round system, but the amounts of hay required per cow were 0, 0, and 4720 lbs dry matter/cow for yearround fall-calving, year-round spring-calving, and minimal land spring-calving cows, respectively.

Evaluation of a Year-Round Grazing System: Winter Progress Report

The winter component of a year-round grazing system involving grazing of corn crop residues followed by grazing stockpiled grass-legume forages was compared at the McNay Research Farm with that of the winter component of a minimal land system that maintained cows in drylot. In the summers of 1995 and 1996, two and one cuttings of hay per year were harvested from two 15-acre fields containing “Johnston” low endophtye tall fescue and red clover. Two cuttings of hay in 1995 and one cutting in 1996 were harvested from two 15-acre fields of smooth bromegrass and red clover. Hay yields were 4,236 and 4,600 pounds of dry matter per acre for the tall fescue-red clover in 1995 and 1996, and 2,239 and 2,300 pounds of dry matter per acre for the smooth bromegrass-red clover in 1995 and 1996. Following grain harvest, four 7.5-acre fields containing corn crop residues were stocked with cows at midgestation at an allowance of 1.5 acres per cow. Forage yields at the initiation of corn crop grazing in 1995 and 1996 were 3,757 and 3,551 pounds of dry matter per acre for corn crop residues. Stockpiled forage yields were 1,748 and 2,912 pounds of dry matter for tall fescue-red clover and 1,880 and 2,187 pounds for smooth bromegrass-red clover. Corn crop residues and stockpiled forages were grazed in a strip stocking system. For comparison, 20 cows in 1995 and 16 cows in 1996 were placed in two drylots simultaneously with initiation of corn crop grazing, where they remained throughout the winter and spring grazing periods. Cows maintained in drylots or grazing corn crop residue and stockpiled forages were supplemented with hay as large round bales to maintain a body condition score of five. In both years, no seasonal differences in body weight and body condition score were observed between grazing cows or cows maintained in drylots, but grazing cows required 85% and 98% less harvested hay in years 1 and 2 than cows in drylot during the winter and spring. Because less hay was needed to maintain grazing cows, excesses of 12,354 and 5,244 pounds of hay dry matter per cow in 1995 and 1996 remained in the year-round grazing system. During corn crop grazing, organic matter yield decreased at 23.5 and 28.8 pounds of organic matter per day from grazed areas of corn crop residues in 1995 and 1996. Organic matter losses due to weathering were 6.8, 10.3, and 12.7 pounds per day in corn crop residue, tall fescue-red clover and smooth bromegrass-red clover in 1995 and 12.1, 10.7, and 12.1 in 1996. Organic matter losses from grazed and ungrazed areas of tall fescue-red clover and smooth bromegrass-red clover during stockpiled grazing were 6.9, 6.9, and 2.1, 2.9 in 1995 and 13.4, 4.3, and +6.9, 4.4 pounds per day in 1996.

Evaluation of Stockpiled Berseem Clover and Brown Midrib Sorghum x Sudangrass as Supplements for Grazed Cornstalks in Beef Cow Wintering Systems

Berseem clover and oats were incorporated into a corncorn- oat/berseem clover rotation in 1994 and 1995. Two cuttings of oat-berseem clover hay were harvested during the summer before forage was allowed to stockpile for winter grazing. In 1995, a brown midrib sorghum x sudangrass hybrid was seeded into a field adjacent to a corn field. After corn grain harvest in 1994 and 1995, Charolais x Angus x Simmental cows in midgestation were allotted to replicated fields containing corn crop residues with no complementary forages at 2.5 acres/cow, or corn crop residues and stockpiled berseem clover (2:1) at 2.5 acres/cow to simultaneously graze, or to a drylot. In 1995, cows were allotted to fields containing corn crop residues and brown midrib sorghum x sudangrass (7:3) at 2.5 acres/cow. Berseem clover had greater concentrations of digestible organic matter and crude protein than corn crop residues at the initiation of grazing, but had a more rapid decrease in digestible organic matter concentration than corn crop residues. Brown midrib sorghum x sudangrass forage also had a higher initial concentration of digestible organic matter, but an equal rate of decrease in digestible organic matter concentration to corn crop residues in ungrazed areas of the field. Cows grazing berseem clover with corn crop residues had greater body condition score increases during the first half of the grazing season than cows grazing corn crop residues without complementary forages. Cows grazing corn crop residues without complementary forages required 2,786 and 1,412 less lb hay per cow than cows maintained in a drylot in 1994 and 1995. In 1994, simultaneous grazing of berseem clover with corn crop residues did not reduce hay feeding more than feeding corn crop residues alone. However, in 1995, grazing berseem clover or brown midrib sorghum x sudangrass with corn crop residues reduced the amount of hay required to maintain cows by 358 and 376 lb hay per cow compared with grazing corn crop residues without complementary forage.

Evaluation of a Year-Round Grazing System: Summer Cow-Calf Progress Report

A comparison was made between two different summer grazing systems at the McNay Research Farm. One system was the summer component of a year-round grazing system, involving the rotational stocking of smooth bromegrass-orchardgrass-birdsfoot trefoil pastures and winter stockpile pastures with cow-calf pairs co-grazing with stocker yearlings at .75 animal units per acre. That system was compared with a minimal land system involving the rotational stocking of smooth bromegrass-orchardgrass-birdsfoot trefoil summer pastures with cow-calf pairs grazing at .64 animal units per acre and hay removal from 25% of the pasture. Stocker yearlings or hay removal were used as management tools to remove excess forage and optimize forage quality. Hay was removed once from three fourths of the winter stockpiled pastures in 1996 (Yr. 1) and all the pasture in 1997 (Yr. 2). One hay removal occurred on one fourth of the allocated summer pastures in Year 1 and one half of the pastures in Year 2. In Year one, cow-calf pairs grazing in the year-round system utilized one fourth of the winter stockpile pastures due to a lack of forage on the summer pastures, whereas in Year 2 cowcalf pairs grazed winter stockpile pastures to remove forage as a second cutting of hay. Cow-calf pairs grazing with hay removal were supplemented with harvested hay for two weeks during the summer of Year 1 due to lack of grazable forage; in Year 2, no supplementation was needed. Grazing system did not affect cow body weight, condition score, or daily calf gain in either year. Growing animal production per acre was affected by grazing system, with the minimal land system having a higher production level in Year 1 and Year 2. The year-round system also produced more net winter forage than did the minimal land system in Year 1. Differences in forage yield and quality were only observed between winter stockpile forages of tall fescue-red clover and smooth bromegrass-red clover and summer pastures during the months of June, July, and August.

Evaluation of Stockpiled Berseem Clover and Brown Midrib Sorghum x Sudangrass as Supplements for Grazed Cornstalks in Beef Cow Wintering Systems

Berseem clover and oats were incorporated into a corn-oat/berseem clover rotation in 1994-1996. Two cuttings of oat-berseem clover hay were harvested during the summer before forage was stockpiled for winter grazing. In 1995, brown midrib sorghum x sudangrass hybrid was seeded into a field adjacent to a corn field. This was repeated in 1996 with a standard sorghum x sudangrass hybrid. After corn harvest in 1994–1996, Charolais x Angus x Simmental cows and heifers in midgestation were allotted to corn crop residue, corn crop residue-berseem clover, and corn crop residue-sorghum x sudangrass fields at 2.5 acres/cow, or to a drylot. Berseem clover had greater concentration of digestible organic matter and crude protein than corn crop residues. Corn crop residue digestible organic matter concentration was lower than berseem clover and the brown midrib sorghum x sudangrass, but was higher than that of the standard sorghum x sudangrass hybrid in 1996. Cows grazing corn crop residues without complementary forages required an average of 2,374 less lb. hay per cow than cows maintained in a drylot in 1994-1996. In 1994 and 1996, simultaneous grazing of berseem clover with corn crop residues did not reduce hay feeding more than feeding corn crop residues alone, yet did significantly reduce the amount of hay needed in 1995 to maintain cows by 358 and 376 lb. hay per cow compared with grazing corn crop residues without complementary forage.

Evaluation of a Year-Round Grazing System: Winter Progress Report

The winter component of a year-round grazing system involving grazing of corn crop residues followed by grazing stockpiled grass legume forages was compared at the McNay Research Farm with that of the winter component of a minimal land system that maintained cows in drylot,. In the summer of 1995, two cuttings of hay were harvested from two 15-acre fields containing “Johnston” endophyte-free tall fescue and red clover, and two cuttings of hay were taken from two 15-acre fields of smooth bromegrass and red clover. Hay yields were 4,236 and 4,600 pounds of dry matter per acre for the tall fescue--red clover and smooth bromegrass--red clover. Following grain harvest four 7.5-acre fields containing corn crop residue were stocked with cows at midgestation at an allowance of 1.5 acres per cow. Forage yields at the initiation of corn crop grazing were 3,766pounds of dry matter per acre for corn crop residue, 1,748 pounds for tall fescue--red clover, and 1,.880 pounds for smooth bromegrass--red clover. Corn crop residues and stockpiled forages were grazed in a strip stocking system. For comparison, 20 cows were placed in two drylots simultaneously to the initiation of corn crop grazing where they remained throughout the winter and spring grazing seasons. Cows maintained in drylot or grazing corn crop residue and stockpiled forages were supplemented with hay as large round bales to maintain a body condition score of five. No seasonal differences in body weight and body condition were observed between grazing cows or cows maintained in drylot, but grazing cows required 87% and 84% less harvested hay than cows in drylot during the winter and spring respectively. Because less hay was needed to maintain grazing cows, an excess of 11,905 and 12,803 pounds of hay dry matter per cow remained in the year-round grazing system. During corn crop grazing, organic matter yield decreased at 27.3 pounds of organic matter per day from grazed areas of corn crop residue. Organic matter losses due to weathering were 9.4, 12.9, and 15.8 pounds per day in corn crop residue, tall fescue-red clover and smooth bromegrass-red clover. Organic matter losses from grazed and ungrazed areas during stockpiled grazing were 7.3 and 6.9 for tall fescue--red clover and 2.1, 2.9 for smooth bromegrass--red clover.

Technical note: Comparison of automated ribosomal intergenic spacer analysis (ARISA) and denaturing gradient gel electrophoresis (DGGE) to assess bacterial diversity in the rumen of sheep

The aim of this study was to compare automated ribosomal intergenic spacer analysis (ARISA) and denaturing gradient gel electrophoresis (DGGE) techniques to assess bacterial diversity in the rumen of sheep. Sheep were fed 2 diets with 70% of either alfalfa hay or grass hay, and the solid (SOL) and liquid (LIQ) phases of the rumen were sampled immediately before feeding (0 h) and at 4 and 8 h postfeeding. Both techniques detected similar differences between forages, with alfalfa hay promoting greater (P < 0.05) bacterial diversity than grass hay. In contrast, whereas ARISA analysis showed a decrease (P < 0.05) of bacterial diversity in SOL at 4 h postfeeding compared with 0 and 8 h samplings, no variations (P > 0.05) over the postfeeding period were detected by DGGE. The ARISA technique showed lower (P < 0.05) bacterial diversity in SOL than in LIQ samples at 4 h postfeeding, but no differences (P > 0.05) in bacterial diversity between both rumen phases were detected by DGGE. Under the conditions of this study, the DGGE was not sensitive enough to detect some changes in ruminal bacterial communities, and therefore ARISA was considered more accurate for assessing bacterial diversity of ruminal samples. The results highlight the influence of the fingerprinting technique used to draw conclusions on factors affecting ruminal bacterial diversity.

Otimização do método lignina brometo de acetila na determinação do teor de lignina em plantas forrageiras e comparação com os métodos lignina detergente ácido e lignina Klason

Técnicas analíticas empregadas para a quantificação do teor de lignina em plantas forrageiras, atualmente em uso, são questionáveis quanto às suas acurácias. O método lignina detergente ácido (LDA), que é um dos métodos mais utilizado em Ciência Animal e Agronomia, apresenta algumas falhas, particularmente devido à parcial solubilização da lignina durante a preparação da fibra em detergente ácido (FDA). A lignina Klason (LK), outro método muito usado, apresenta o inconveniente de mensurar a proteína da parede celular como sendo lignina. Em ambos os procedimentos recomenda-se também mensurar cinzas nos resíduos de lignina. A quantificação da concentração de lignina pelo método espectrofotométrico lignina brometo de acetila (LBA) vem ganhando interesse de pesquisadores no Brasil e no exterior. Nesta metodologia, a lignina da planta contida na preparação parede celular (PC) é solubilizada numa solução a 25% de brometo de acetila em ácido acético e a absorbância mensurada é com luz UV a 280 nm. O valor da absorbância é inserido numa equação de regressão e a concentração de lignina é obtida. Para que esta técnica analítica seja mais aceita pelos pesquisadores, ela deve ser, obviamente, convincente e atrativa. O presente trabalho analisou alguns parâmetros relacionados à LBA em 7 gramíneas e 6 leguminosas, em dois estádios de maturidade. Dentre as diferentes temperaturas de pré-secagem, os resultados indicaram que os procedimentos de 55°C com ventilação e liofilização podem ser utilizados com a mesma eficácia. As temperaturas de 55°C sem ventilação e 80°C sem ventilação não são recomendadas, pois aumentaram os valores de FDA e LDA, possivelmente devido ao surgimento de artefatos de técnica como os compostos de Maillard. No método LBA os valores menores das amostras de leguminosas chamaram a atenção e colocaram em questão se a lignina destas plantas seria menos solúvel no reagente brometo de acetila. Dentre algumas alterações na metodologia da técnica LBA, a utilização do moinho de bolas (para diminuir o tamanho particular) nas amostras de PC não mostrou efeito; a hipótese era melhorar a solubilização da lignina usando partículas menores. O uso de um ultrasonicador, que aumenta a vibração das moléculas e assim, facilitaria a solubilização da lignina no reagente brometo de acetila, melhorou a solubilização da lignina em cerca de 10%, tanto nas gramíneas como nas leguminosas. Foi acoplado um ensaio biológico como referência, a degradabilidade in vitro da matéria seca (DIVMS); e como a lignina está intimamente associada à estrutura fibrosa da parede celular, também foi feito um ensaio de degradabilidade in vitro da fibra em detergente neutro (DIVFDN). Os resultados confirmaram o efeito da maturidade, reduzindo a degradabilidade nas plantas mais maduras, e que o teor de lignina de leguminosas é realmente inferior ao de gramíneas. Os resultados de degradabilidade apresentaram coeficientes de correlação mais elevados com o método LBA, quando foi empregada a técnica do ultrasom; o método LK mostrou os menores coeficientes. Também testou-se, com sucesso, a utilização da FDN, como preparação fibrosa, ao invés de PC. A razão é simples: enquanto que a FDN é amplamente conhecida, a preparação PC não o é. Inquestionável que esta manobra facilitará substancialmente a divulgação desse método, tornando-a mais aceitável pela comunidade científica