Réal Michaud

Alfalfa Cut at Sundown and Harvested as Baleage Improves Milk Yield of Late-Lactation Dairy Cows

Alfalfa (Medicago sativa L.) cut at sundown has been shown to contain greater concentration of total nonstructural carbohydrates (TNC) than that cut at sunup. Fourteen multiparous (8 ruminally cannulated) and 2 primiparous lactating dairy cows were randomly assigned to 2 treatments in a crossover design (2 periods of 24 d) to investigate the effects of alfalfa daytime cutting management on ruminal metabolism, nutrient digestibility, N balance, and milk yield. Half of each alfalfa field (total of 3 fields) was cut at sundown (PM) after a sunny day, whereas the second half was cut at sunup (AM) on the following day. Both PM and AM cuts were field-wilted and harvested as baleage (531 +/- 15.0 g of dry matter/kg of fresh matter). Bales (PM and AM) were ranked according to their concentrations of TNC, paired, and each pair of PM and AM baleages was then assigned to each experimental day (total of 48 d). The difference in TNC concentration between PM and AM baleages fed during the 10 d of data and sample collection varied from -10 to 50 g/kg of dry matter. Each pair of baleage was fed ad libitum to cows once daily with no concentrate. Ruminal molar proportion of acetate and total volatile fatty acid concentration were greater in animals fed the AM baleage, whereas the proportion of valerate was greater with PM baleage; no other significant changes in ruminal molar proportions of volatile fatty acids were observed between forage treatments. Digestible organic matter intake, organic matter digestibility, and plasma Lys concentration were significantly greater in cows fed PM alfalfa, suggesting that more nutrients were available for milk synthesis. Significantly lower body weight gain and retained N as a proportion of N intake were observed in cows fed PM alfalfa, thus suggesting that nutrients were channeled to milk synthesis rather than to body reserves. Intake of dry matter (+1.0 kg/d), and yields of milk (+1.0 kg/d), milk fat (+70 g/d), and milk protein (+40 g/d) were significantly greater in cows fed PM vs. AM alfalfa. Concentration of milk urea N and excretion of urea N as a proportion of total urinary N were significantly reduced, and milk N efficiency was increased when feeding PM vs. AM alfalfa, indicating an improvement in N utilization. Increasing the TNC concentration of alfalfa by shifting forage cutting from sunup to sundown improved N utilization and milk production in late-lactation dairy cows.

Alfalfa cut at sundown and harvested as baleage increases bacterial protein synthesis in late-lactation dairy cows.

Alfalfa (Medicago sativa L.) cut at sundown (p.m.) has been shown to have a greater concentration of total nonstructural carbohydrates (TNC) than when cut at sunup (a.m.). Eight ruminally cannulated Holstein cows that were part of a larger lactation trial were used in a crossover design (24-d periods) to investigate the effects of alfalfa cutting time on digestibility and omasal flow of nutrients. Alfalfa was cut at sundown or sunup, field-wilted, and harvested as baleage (530 +/- 15.0 g of dry matter/kg of fresh matter). The difference in TNC concentration between p.m. and a.m. alfalfa within each pair of bales fed daily during the 10 d of data and sample collection varied from -10 to 50 g/ kg of dry matter. Each pair of bales was fed for ad libitum intake to cows once daily with no concentrate. During the 3 d of omasal sampling, intake (+0.8 kg/d) and omasal flow of organic matter (OM; +0.42 kg/d) tended to be greater when cows were fed p.m. vs. a.m. alfalfa, but no differences were found for ruminal and postruminal digestion of this nutrient. Similarly, N apparently digested ruminally and postruminally did not differ when feeding p.m. vs. a.m. alfalfa. However, N truly digested in the rumen, as a proportion of N intake, was significantly greater in cows fed p.m. (79%) vs. a.m. alfalfa (74%), thus suggesting that longer wilting time of alfalfa cut at sundown increased forage proteolysis. Supply of rumen-degradable protein did not change (2,716 g/d) when averaged across treatments, whereas omasal flow of non-NH(3) nonbacterial N was significantly decreased (-29 g/d) when feeding p.m. vs. a.m. alfalfa. Omasal flow of total bacterial non-NH(3)-N (NAN) increased (+21 g/d) significantly when cows were fed p.m. vs. a.m. alfalfa possibly because bacteria from cows fed p.m. alfalfa captured significantly more NH(3) than those from cows fed a.m. alfalfa. Therefore, greater availability of fermentable energy as TNC appears to increase the capacity of microbes to uptake NH(3)-N and convert it to microbial protein. Enhanced OM intake can also explain the observed increase in bacterial protein synthesis with p.m. alfalfa. Efficiency of bacterial protein synthesis, expressed on a fermented OM basis or as grams of bacterial NAN per gram of rumen-degradable N, did not differ between p.m. and a.m. alfalfa. Conversely, bacterial efficiency, as grams of bacterial NAN per gram of N intake, was significantly increased when cows were fed p.m. baleage. No significant difference between forage treatments was found for the omasal flow of total AA from omasal true digesta, suggesting no benefit of daytime cutting management on the passage of total AA to the lower gastrointestinal tract. Enhancing energy intake and TNC concentration of alfalfa by shifting forage cutting from sunup to sundown increased protein synthesis and NH(3) uptake by ruminal bacteria indicating an improvement in N utilization.

Winter damage to perennial forage crops in eastern Canada : Causes, mitigation, and prediction

Harsh winter climate results in frequent losses of stands and yield reduction in many forage-growing areas of Canada and other parts of the world. Climatic conditions and crop management both affect the winter survival of perennial forage crops. In this review, we present the main causes of winter damage in eastern Canada and we discuss crop management practices that help mitigate the risks of losses. Predictive tools available to assess the risks of winter damage both spatially and temporally are also presented. Our understanding of the causes of winter damage and of the plant adaptation mechanisms to winter stresses, particularly the role of N and C organic reserves, has improved. Forage species commonly grown in eastern Canada differ in their tolerance to subfreezing temperatures and to anoxia caused by the presence of ice on fields. Some improvement in winter hardiness of forage legume species has been achieved through breeding in eastern Canada but new technologies based on laboratory freezing tests ...

Improving the nutritive value of timothy through management and breeding

Timothy (Phleum pratense L.) is a widely grown fo rage grass species in cool and humid regions of the world including northeastern and northwestern North America, Nordic countries, Russia, and Japan. The nutritive value of timothy decreases with time, phenological development, and increasing forage yield. This review paper summarizes methods of controlling or improving the nutritive value of timothy through management practices and genetic selection, while keeping in mind the importance of forage yield. Consequently, the nutritive value of timothy is considered in relation to the accumulation of forage yield. The ecophysiological basis for the decrease in nutritive value with increasing forage yield during a growth cycle is presented with the assumption that the forage is made of two components: metabolic and structural. The decrease in the proportion of the metabolic component with increasing forage yield reduces the nutritive value of timothy. The nutritive value is also affected by the N concentration ...

Genetic transformation of commercial breeding lines of alfalfa (Medicago sativa)

Bio-engineering technologies are now routinely used for the genetic improvement of many agricultural crops. However, breeding lines of Medicago sativa are not easily amenable to genetic transformation and therefore cannot benefit from the molecular tools that have been developed for genetic manipulations. This paper describes a strategy that has been developed to transfer DNA into commercially important breeding lines of winter-hardy alfalfa via Agrobacterium infection. Three highly regenerative genotypes have been selected from ca 1000 genotypes within 11 breeding lines. They have been used as basic material for an extensive genetic transformation trial. Combinations of genotypes (11.9, 8.8, 1.5) expression vectors (pGA482, pGA643, pBibKan) and bacterial strains (C58, A281, LBA4404) were tested for their ability to produce stable transgenic material. Putative transgenic plantlets were further screened by nptII-specific PCR amplification, Southern hybridization and recallusing assays. One genotype (1.5) gave only one transformant out of 432 individual trials. With the two other genotypes, efficiency of transformation (kanamycin-resistant calluses obtained/explant tested) ranged from 0 to 0.92 depending on the strain/vector combination used. Statistical interactions underline the possibility of obtaining good genotype-strain-vector combinations for alfalfa transformation. Predicted transformation probability indicates that with strain LBA4404 containing the vector pGA482 and genotype 11.9, transformation efficiency is above 60% and 10% or more of the calluses retain embryogenic potential. PCR amplification and Southern hybridization of randomly chosen regenerated plantlets demonstrated that all embryos developing on 50 μg ml-1 kanamycin had a stable genomic insertion of nptII. Sexual crosses with untransformed genotypes showed that segregation of the transgenic trait followed Mendelian heredity.

SRAP polymorphisms associated with superior freezing tolerance in alfalfa (Medicago sativa spp. sativa)

Sequence-related amplified polymorphism (SRAP) analysis was used to uncover genetic polymorphisms among alfalfa populations recurrently selected for superior tolerance to freezing (TF populations). Bulk DNA samples (45 plants/bulk) from each of the cultivar Apica (ATF0), and populations ATF2, ATF4, ATF5, and ATF6 were evaluated with 42 different SRAP primer pairs. Several polymorphisms that progressively intensified or decreased with the number of recurrent cycles were identified. Four positive polymorphisms (F10-R14, Me4-R8, F10-R8 and F11-R9) that, respectively, yielded 540-, 359-, 213-, and 180-bp fragments were selected for further analysis. SRAP amplifications with genotypes within ATF populations confirmed that the polymorphisms identified with bulk DNA samples were reflecting changes in the frequency of their occurrence in response to selection. In addition, the number of genotypes cumulating multiple polymorphisms markedly increased in response to recurrent selection. Independent segregation of the four SRAP polymorphisms suggests location at unlinked loci. Homology search gave matches with BAC clones from syntenic Medicago truncatula for the four SRAP fragments. Analysis of the relationship with low temperature tolerance showed that multiple SRAP polymorphisms are more frequent in genotypes that maintain superior regrowth after freezing. These results show that SRAP analysis of bulk DNA samples from recurrent selections is an effective approach for the identification of genetic polymorphisms associated with quantitative traits in allogamous species. These polymorphisms could be useful tools for indirect selection of freezing tolerance in alfalfa.

Association Mapping of Biomass Yield and Stem Composition in a Tetraploid Alfalfa Breeding Population

Alfalfa (Medicago sativa L.), an important forage crop that is also a potential biofuel crop, has advantages of high yield, high lignocellulose concentration in stems, and has low input costs. In this study, we investigated population structure and linkage disequilibrium (LD) patterns in a tetraploid alfalfa breeding population using genome‐wide simple sequence repeat (SSR) markers and identified markers related to yield and cell wall composition by association mapping. No obvious population structure was found in our alfalfa breeding population, which could be due to the relatively narrow genetic base of the founders and/or due to two generations of random mating. We found significant LD (p < 0.001) between 61.5% of SSR marker pairs separated by less than 1 Mbp. The observed large extent of LD could be explained by the effect of bottlenecking and selection or the high mutation rates of SSR markers. Total marker heterozygosity was positively related to biomass yield in each of five environments, but no relationship was noted for stem composition traits. Of a total of 312 nonrare (frequency >10%) alleles across the 71 SSR markers, 15 showed strong association (p < 0.005) with yield in at least one of five environments, and most of the 15 alleles were identified in multiple environments. Only one allele showed strong association with acid detergent fiber (ADF) and one allele with acid detergent lignin (ADL). Alleles associated with traits could be directly applied in a breeding program using marker‐assisted selection. However, based on our estimated LD level, we would need about 1000 markers to explore the whole alfalfa genome for association between markers and traits.

Effects of nonstructural carbohydrate concentration in alfalfa on fermentation and microbial protein synthesis in continuous culture

Insufficient readily fermentable energy combined with extensive degradation of proteins in alfalfa (Medicago sativa L.) may result in poor forage N utilization by ruminants. Using the inherent genetic variability and differences between harvests, our objective was to compare the effect of contrasting concentrations of nonstructural carbohydrates (NSC) in alfalfa on rumen fermentation and microbial protein synthesis. Individual genotypes of the alfalfa cultivar AC Caribou grown near Québec City, Québec, Canada, were harvested at the vegetative and early flowering stages, dried at 55 degrees C, ground, and analyzed for soluble carbohydrates (fructose + sucrose + glucose + pinitol) and starch. Approximately 20 genotypes having, respectively, the highest and lowest NSC concentrations were pooled to constitute 2 contrasted 1-kg forage samples. Samples of high- (17.9% DM) and low- (7.4% DM) NSC alfalfa were respectively allocated to separate dual-flow fermenters in a completely randomized design with 3 replications. Rumen inoculum was obtained from 4 ruminally fistulated cows in early lactation that were fed a TMR with a 50:50 forage to concentrate ratio. A 10-d incubation period was used, with the first 6 d serving as an adaptation period followed by 4 d of sampling with solid and liquid dilution rates in the fermenters set at approximately 2.0 and 4.3%/h, respectively. High versus low NSC concentration in alfalfa significantly enhanced the apparent digestibility of OM (59.1% for high-NSC alfalfa vs. 54.4% for low-NSC alfalfa) and DM (60.0 vs. 54.3%) and the true digestibility of DM (74.1 vs. 64.7%). Increasing NSC concentration in alfalfa (high vs. low) significantly decreased ruminal pH (6.85 vs. 7.08) and NH(3)-N concentration (26.0 vs. 33.6 mg/dL) and increased total VFA concentration (94.9 vs. 83.0mM). Molar proportions of acetate, isobutyrate, and isovalerate significantly decreased, whereas molar proportions of propionate and butyrate significantly increased with high-NSC alfalfa, resulting in a more glucogenic fermentation. More importantly, microbial N flow (263 vs. 230 mg/d) and bacterial N efficiency (41.1 vs. 29.6% of available N), measured using (15)N as a microbial marker, both significantly increased with the high-NSC alfalfa. These results indicate that increasing the concentration of NSC in alfalfa promotes a glucogenic fermentation and enhances microbial N synthesis in the rumen.

Near-infrared reflectance spectroscopy prediction of neutral detergent-soluble carbohydrates in timothy and alfalfa

Carbohydrates in forage crops can be divided into neutral detergent-insoluble fiber and neutral detergent-soluble carbohydrates (NDSC); the latter includes organic acids (OA), total ethanol:water-soluble carbohydrates (TESC), starch, and neutral detergent-soluble fiber (NDSF). The accurate and efficient estimation of NDSC in forage crops is essential for improving the performance of dairy cattle. In the present study, visible and near-infrared reflectance spectroscopy (NIRS) were applied to evaluate the feasibility of predicting OA, TESC, starch, NDSF, NDSC, and all related constituents used to calculate these 5 carbohydrate fractions in timothy and alfalfa. Forage samples (n = 1,008) of timothy and alfalfa were taken at the first and second harvests at 2 sites in 2007; samples were dried, ground, and then scanned (400 to 2,500 nm) using an NIRSystems 6500 monochromator. A calibration (n = 60) and a validation (n = 15) set of samples were selected for each species and then chemically analyzed. Concentrations of TESC and NDSC in timothy, as well as starch in alfalfa, were successfully predicted, but many other carbohydrate fractions were not predicted accurately when calibrations were performed using single-species sample sets. Both sets of samples were combined to form new calibration (n = 120) and validation (n = 30) sets of alfalfa and timothy samples. Calibration and validation statistics for the combined sets of alfalfa and timothy samples indicated that TESC, starch, and NDSC were predicted successfully, with coefficients of determination of prediction of 0.92, 0.89, and 0.93, and a ratio of prediction to deviation (RPD) of 3.3, 3.1, and 3.6, respectively. The NDSF prediction was classified as moderately successful The NIRS prediction of OA was unsuccessful All related constituents were predicted successfully by NIRS except ethanol-insoluble residual OM, with Our results confirm the feasibility of using NIRS to predict NDSC, its fractions, and other related constituents, except for OA and ethanol-insoluble residual OM, in timothy and alfalfa forage samples.

Leaf and stem dry matter digestibility and ruminal undegradable proteins of alfalfa cultivars

Increased dry matter digestibility and ruminal undegradable protein (RUP) concentration are traits highly ranked to improve the nutritive value of forage legumes. Whole-plant, leaf, and stem samples of 27 alfalfa cultivars grown in eastern Canada were analysed for total nitrogen (TN), in vitro RUP, and in vitro dry matter digestibility (IVDMD). The objectives were to determine if significant variation exists among these cultivars, and to establish the relationship between nutritive value parameters of leaves and stems, and those of whole plants. Cultivars were seeded in triplicate on 2 consecutive years and evaluated during the 2 subsequent production years with two harvests per year. There were significant differences among cultivars in IVDMD of stems and plants in spring growth and summer regrowth. Cultivar differences in leaf IVDMD were significant only in spring growth, but this variation was negligible in practical terms. Cultivars with high forage IVDMD also had high stem IVDMD and this relationship...