Carbon assimilation, herbage accumulation, nutritive value, and grazing efficiency of Mulato II brachiariagrass under continuous stocking

Grazed pastures are the backbone of the Brazilian livestock industry and grasses of the genus Brachiaria (syn. Urochloa) are some of most used tropical forages in the country. Although the dependence on the forage resource is high, grazing management is often empirical and based on broad and non-specific guidelines. Mulato II brachiariagrass (Convert HD 364, Dow AgroSciences, São Paulo, Brazil) (B. brizantha × B. ruziziensis × B. decumbens), a new Brachiaria hybrid, was released as an option for a broad range of environmental conditions. There is no scientific information on specific management practices for Mulato II under continuous stocking in Brazil. The objectives of this research were to describe and explain variations in carbon assimilation, herbage accumulation (HA), plant-part accumulation, nutritive value, and grazing efficiency (GE) of Mulato II brachiariagrass as affected by canopy height and growth rate, the latter imposed by N fertilization rate, under continuous stocking. An experiment was carried out in Piracicaba, SP, Brazil, during two summer grazing seasons. The experimental design was a randomized complete block, with a 3 x 2 factorial arrangement, corresponding to three steady-state canopy heights (10, 25 and 40 cm) maintained by mimicked continuous stocking and two growth rates (imposed as 50 and 250 kg N ha-1 yr-1), with three replications. There were no height × N rate interactions for most of the responses studied. The HA of Mulato II increased linearly (8640 to 13400 kg DM ha-1 yr-1), the in vitro digestible organic matter (IVDOM) decreased linearly (652 to 586 g kg-1), and the GE decreased (65 to 44%) as canopy height increased. Thus, although GE and IVDOM were greatest at 10 cm height, HA was 36% less for the 10- than for the 40-cm height. The leaf carbon assimilation was greater for the shortest canopy (10 cm), but canopy assimilation was less than in taller canopies, likely a result of less leaf area index (LAI). The reductions in HA, plant-part accumulation, and LAI, were not associated with other signs of stand deterioration. Leaf was the main plant-part accumulated, at a rate that increased from 70 to 100 kg DM ha-1 d-1 as canopy height increased from 10 to 40 cm. Mulato II was less productive (7940 vs. 13380 kg ha-1 yr-1) and had lesser IVDOM (581 vs. 652 g kg-1) at the lower N rate. The increase in N rate affected plant growth, increasing carbon assimilation, LAI, rates of plant-part accumulation (leaf, stem, and dead), and HA. The results indicate that the increase in the rate of dead material accumulation due to more N applied is a result of overall increase in the accumulation rates of all plant-parts. Taller canopies (25 or 40 cm) are advantageous for herbage accumulation of Mulato II, but nutritive value and GE was greater for 25 cm, suggesting that maintaining ∼25-cm canopy height is optimal for continuously stocked Mulato II.

Massa seca e composição bromatológica de quatro espécies de braquiárias semeadas na linha ou a lanço, em consórcio com milho no sistema plantio direto na palha

The objective of this research was to evaluate the dry mass yield and chemical composition of four Brachiaria species in different options for sowing, exclusively or in intercrop with corn crop, under a no-tillage system. The experiment was carried out during the growing seasons of 2006 at FEPE (FE/Unesp, Ilha Solteira Campus) located in Selvíria, Mato Grosso do Sul State, Brazil. The soil of the experimental area was classified as distroferric Red Latosol (Oxisol). The experimental design was in randomized blocks, in a factorial scheme (4 × 4), with five replications. The treatments consisted of four Brachiaria species (Brachiaria brizantha cv. Marandu, Brachiaria decumbens, Brachiaria ruziziensis and Mulato II grass) grown in rows and spread on total area, exclusively or intercropped simultaneously with corn crop sowing. The study evaluated the dry mass yield and total digestible nutrients, crude protein, ash, neutral detergent fiber, acid detergent fiber, hemicelluloses, cellulose and lignin content of forage. The spread on total area intercrop of forages with corn crop proved to be viable by presenting similar dry mass yield to exclusive sowing arrangements, conversely to what happened with intercrop in row of corn crop, which decreased such yield. Brachiaria ruziziensis showed superior chemical composition and the intercrops increased energy and crude protein contents, and decreased cell wall components.

Evaluación de cultivares a pequeña escala, su adaptabilidad y respuesta a enmiendas orgánicas, en la microcuenca Orocuina, Somoto, 2011

Con el propósito de evaluar rendimientos y adaptabilidad de variedades mejoradas en granos básicos (maíz y frijol), hortalizas (cebolla) y pastos, ante las frecuentes sequías y su efecto a la mejora de la calidad de suelo en la microcuenca Orocuina, municipio de Somoto; se establecieron en tres fincas representativas del área un total de nueve parcelas ó ensayos productivos en sitios que oscilan entre 1 320 y 7 026 m2, en época de postrera 2011. Las variedades utilizadas fueron maíz Nutrader, maíz criollo Catacama, frijol INTA-Rojo y frijol balín como criollo, cebolla Orlando, pasto Mulato II y gamba. Previo al establecimiento de los ensayos, se seleccionaron localmente un grupo de indicadores sencillos entre los cuales están; el contenido de materia orgánica en el suelo, tasa de infiltración de agua en el suelo, textura, pendiente del terreno, profundidad del suelo, piedras en la superficie, espesor orgánico, pH y fósforo disponible. Los cálculos indican que el contenido de materia orgánica comprende niveles medios para las tres fincas bajo estudio, este indicador es el más importante ya que permite evaluar la calidad de suelos en vista de la influencia positiva en las propiedades y funciones básicas del suelo. Los niveles de fósforo disponible van de pobre a bueno, presenta suelos óptimos para la agricultura, textura es arcillosa, franca arcillo arenosa y franco arcillo limosa; sin embargo, estos suelos tienen potencial para establecer una alternativa que permita incrementar los rendimientos de los cultivos, mediante prácticas de manejo adecuado como obras de conservación de suelos garantizando de esta manera la sostenibilidad de este recurso. Los resultados obtenidos indicaron una respuesta propia a diferentes aplicaciones de las fuentes de fertilizantes, la fertilización sintética (urea) y abono orgánico (lombrihumus), los rendimientos obtenidos fueronde 1 6 82kg ha-1 (maíz Nutrader) y 1 954 kg ha-1(maíz Catacama);1045 kg ha-1 (frijol INTA-Rojo) y 1 136 kg ha 1 (frijol Balín); para el cultivo de Cebolla Orlando en la fertilización sintética 14 544 kg ha-1, compuesta 8 954 kg ha-1 y sin fertilizar 6 079 kg ha-1 respectivamente. Para la fertilización compuesta (18-46-0 más lombrihumus),20 271 kg ha-1 (pasto mulato II)y 15 180 kg ha-1 (pasto gamba) de materia seca. El análisis económico muestra que el cultivo de pasto gamba, fertilizado 18-46-0 y lombrihumus obtuvo los mayores beneficios económicos con C$ 2.69, donde se recupera el córdoba invertido más 1.69 córdoba adicional de ganancias.

Produtividade de grãos de milho e massa seca de braquiárias em consórcio no sistema de integração lavoura-pecuária

Objetivou-se avaliar as produtividades de grãos de milho e massa seca de braquiárias em duas modalidades de consórcio em sistema de integração lavoura-pecuária no período de inverno-primavera em região do Cerrado. O experimento foi conduzido no ano de 2006, na Fazenda de Ensino, Pesquisa e Extensão (FEPE), pertencente à Faculdade de Engenharia (FE/UNESP - Campus de Ilha Solteira), localizada no município de Selvíria, MS. O delineamento experimental utilizado foi de blocos casualizados, em esquema fatorial 4x2, com cinco repetições. Os tratamentos constituíram-se de quatro espécies de braquiárias (Brachiaria brizantha cv. 'Marandu', Brachiaria decumbens, Brachiaria ruziziensis e Brachiaria híbrido cv. 'Mulato II') consorciadas na linha e a lanço no momento da semeadura do milho. Avaliaram-se os componentes da produção, a produtividade do milho, bem como a massa seca das braquiárias após a colheita do milho. As forrageiras consorciadas a lanço, com destaque para a Brachiaria ruziziensis proporcionaram menor desenvolvimento das plantas de milho e menores valores dos componentes da produção, bem como da produtividade de grãos. Apesar de satisfatórias produtividades de massa seca (acima de 2.500kg ha-1), com exceção da Brachiaria brizantha, as demais espécies consorciadas a lanço foram superiores, com destaque para a Brachiaria decumbens e a Brachiaria ruziziensis que apresentaram maior adaptabilidade e produtividade de forragem no consórcio com milho em sistema de integração lavoura-pecuária.

Weed suppression in the formation of brachiarias under three sowing methods

The success of conservation systems such as no-till depends on adequate soil cover throughout the year, which is possible through the use of cover crops. For this purpose the species belonging to the genus Urochloa has stood out by virtue of its hardiness and tolerance to drought. Aiming ground cover for the no-till system, the objective was to evaluate the establishment of two species of the genus Urochloa, in three sowing methods, in the weed suppression and the sensitivity of these forages to glyphosate. The study design was a randomized block with a 2 x 3 x 3 factorial arrangement, in which factor A was composed of Urochloa ruziziensis and Urochloa hybrid CIAT 36087 cv. Mulato II, factor B was formed by sowing methods: sown without embedding, sown with light embedding and sown in rows, and factor C was composed of three doses of glyphosate (0.975, 1.625 and 2.275 kg ha(-1) of acid equivalent). For determination of weed suppression, assessment of biomass yield and soil cover was performed, by brachiaria and weeds, at 30, 60, 90, 120 and 258 days after sowing. Visual assessment of the desiccation efficiency at 7 and 14 days after herbicide application was performed. It is concluded that embedding Urochloa seeds stands out in relation to sowing in the soil surface. Urochloa ruziziensis is more efficient in the dry weight yield, weed suppression, in addition to being more sensitive to glyphosate herbicide.

Investigation of organic xenobiotic transfers, partitioning and processing in air-soil-plant systems using a microcosm apparatus. Part II:comparing the fate of chlorobenzenes in grass planted soil

A microcosm system was used to investigate and compare transfers of 14C labeled-1,2-dichlorobenzene (DCB), 1,2,4-trichlorobenzene (TCB) and hexachlorobenzene (HCB) in an air-soil-plant system using single grass tillers planted into spiked soil. This study was the second phase of a development investigation for eventual study of a range of xenobiotic pollutants. Recoveries from the system were excellent at >90%. The predominant loss pathway for 14C labeled-1,2-DCB and 1,2,4-TCB was volatilisation with 85% and 76% volatilisation of parent compound and volatile metabolites over 5 weeks respectively. Most of the added label in the hexachlorobenzene spiked system remained in soil. Mineralisation was

Trans-disciplinary research in synthesis of grass pollen aerobiology and its importance for respiratory health in Australasia

Grass pollen is a major trigger for allergic rhinitis and asthma, yet little is known about the timing and levels of human exposure to airborne grass pollen across Australasian urban environments. The relationships between environmental aeroallergen exposure and allergic respiratory disease bridge the fields of ecology, aerobiology, geospatial science and public health. The Australian Aerobiology Working Group comprised of experts in botany, palynology, biogeography, climate change science, plant genetics, biostatistics, ecology, pollen allergy, public and environmental health, and medicine, was established to systematically source, collate and analyse atmospheric pollen concentration data from 11 Australian and six New Zealand sites. Following two week-long workshops, post-workshop evaluations were conducted to reflect upon the utility of this analysis and synthesis approach to address complex multidisciplinary questions. This Working Group described i) a biogeographically dependent variation in airborne pollen diversity, ii) a latitudinal gradient in the timing, duration and number of peaks of the grass pollen season, and iii) the emergence of new methodologies based on trans-disciplinary synthesis of aerobiology and remote sensing data. Challenges included resolving methodological variations between pollen monitoring sites and temporal variations in pollen datasets. Other challenges included “marrying” ecosystem and health sciences and reconciling divergent expert opinion. The Australian Aerobiology Working Group facilitated knowledge transfer between diverse scientific disciplines, mentored students and early career scientists, and provided an uninterrupted collaborative opportunity to focus on a unifying problem globally. The Working Group provided a platform to optimise the value of large existing ecological datasets that have importance for human respiratory health and ecosystems research. Compilation of current knowledge of Australasian pollen aerobiology is a critical first step towards the management of exposure to pollen in patients with allergic disease and provides a basis from which the future impacts of climate change on pollen distribution can be assessed and monitored.

Grass pollen allergens globally: The contribution of subtropical grasses to burden of allergic respiratory diseases

Grass pollens of the temperate (Pooideae) subfamily and subtropical subfamilies of grasses are major aeroallergen sources worldwide. The subtropical Chloridoideae (e.g. Cynodon dactylon; Bermuda grass) and Panicoideae (e.g. Paspalum notatum; Bahia grass) species are abundant in parts of Africa, India, Asia, Australia and the Americas, where a large and increasing proportion of the world's population abide. These grasses are phylogenetically and ecologically distinct from temperate grasses. With the advent of global warming, it is conceivable that the geographic distribution of subtropical grasses and the contribution of their pollen to the burden of allergic rhinitis and asthma will increase. This review aims to provide a comprehensive synthesis of the current global knowledge of (i) regional variation in allergic sensitivity to subtropical grass pollens, (ii) molecular allergenic components of subtropical grass pollens and (iii) allergic responses to subtropical grass pollen allergens in relevant populations. Patients from subtropical regions of the world show higher allergic sensitivity to grass pollens of Chloridoideae and Panicoideae grasses, than to temperate grass pollens. The group 1 allergens are amongst the allergen components of subtropical grass pollens, but the group 5 allergens, by which temperate grass pollen extracts are standardized for allergen content, appear to be absent from both subfamilies of subtropical grasses. Whilst there are shared allergenic components and antigenic determinants, there are additional clinically relevant subfamily-specific differences, at T- and B-cell levels, between pollen allergens of subtropical and temperate grasses. Differential immune recognition of subtropical grass pollens is likely to impact upon the efficacy of allergen immunotherapy of patients who are primarily sensitized to subtropical grass pollens. The literature reviewed herein highlights the clinical need to standardize allergen preparations for both types of subtropical grass pollens to achieve optimal diagnosis and treatment of patients with allergic respiratory disease in subtropical regions of the world. © 2014 John Wiley & Sons Ltd.

Unique and cross-reactive T cell epitope peptides of the major bahia grass pollen allergen, pas n 1

Background Bahia grass pollen (BaGP) is a major cause of allergic rhinitis. Subcutaneous allergen-specific immunotherapy is effective for grass pollen allergy, but is unsuitable for patients with moderate to severe asthma due to the risk of anaphylaxis. T cell-reactive but IgE nonreactive peptides provide a safer treatment option. This study aimed to identify and characterize dominant CD4+ T cell epitope peptides of the major BaGP allergen, Pas n 1. Methods Pas n 1-specific T cell lines generated from the peripheral blood of BaGP-allergic subjects were tested for proliferative and cytokine response to overlapping 20-mer Pas n 1 peptides. Cross-reactivity to homologous peptides from Lol p 1 and Cyn d 1 of Ryegrass and Bermuda grass pollen, respectively, was assessed using Pas n 1 peptide-specific T cell clones. MHC class II restriction of Pas n 1 peptide T cell recognition was determined by HLA blocking assays and peptide IgE reactivity tested by dot blotting. Results Three Pas n 1 peptides showed dominant T cell reactivity; 15 of 18 (83%) patients responded to one or more of these peptides. T cell clones specific for dominant Pas n 1 peptides showed evidence of species-specific T cell reactivity as well as cross-reactivity with other group 1 grass pollen allergens. The dominant Pas n 1 T cell epitope peptides showed HLA binding diversity and were non-IgE reactive. Conclusions The immunodominant T cell-reactive Pas n 1 peptides are candidates for safe immunotherapy for individuals, including those with asthma, who are allergic to Bahia and possibly other grass pollens.

Influence of harvesting strategy on nutrient supply and production of dairy cows consuming diets based on grass and red clover silage

The main objective of this thesis was to elucidate the effects of regrowth grass silage and red clover silage on nutrient supply and milk production of dairy cows as compared with primary growth grass silages. In the first experiment (publication I), two primary growth and four regrowth grass silages were harvested at two stages of growth. These six silages were fed to 24 lactating dairy cows with two levels of concentrate allowance. Silage intake and energy corrected milk yield (ECM) responses, and the range in these response variables between the diets, were smaller when regrowth silages rather than primary growth silages were fed. Milk production of dairy cows reflected the intake of metabolizable energy (ME), and no differences in the ME utilization were found between the diets based on silages harvested from primary growth and regrowth. The ECM response to increased concentrate allowance was, on average, greater when regrowth rather than primary growth silages were fed. In the second experiment (publication II), two silages from primary growth and two from regrowth used in I were fed to rumen cannulated lactating dairy cows. Cows consumed less feed dry matter (DM), energy and protein, and produced less milk, when fed diets based on regrowth silages rather than primary growth silages. Lower milk production responses of regrowth grass silage diets were mainly due to the lower silage DM intake, and could not be accounted for by differences in energy or protein utilization. Regrowth grass silage intake was not limited due to neutral detergent fibre (NDF) digestion or rumen fill or passage kinetics. However, lower intake may be at least partly attributable to plant diseases such as leaf spot infections, dead deteriorating material or abundance of weeds, which are all higher in regrowth compared with primary growth, and increase with advancing regrowth. In the third experiment (publications III and IV), red clover silages and grass silages harvested at two stages of growth, and a mixed diet of red clover and grass silages, were fed to five rumen cannulated lactating dairy cows. In spite of the lower average ME intake for red clover diets, the ECM production remained unchanged suggesting more efficient utilisation of ME for red clover diets compared with grass diets. Intake of N, and omasal canal flows of total non-ammonia N (NAN), microbial and non-microbial NAN were higher for red clover than for grass silage diets, but were not affected by forage maturity. Delaying the harvest tended to decrease DM intake of grass silage and increase that of red clover silage. The digestion rate of potentially digestible NDF was faster for red clover diets than for grass silage diets. Delaying the harvest decreased the digestion rate for grass but increased it for red clover silage diets. The low intake of early-cut red clover silage could not be explained by silage digestibility, fermentation quality, or rumen fill but was most likely related to the nutritionally suboptimal diet composition because inclusion of moderate quality grass silage in mixed diet increased silage DM intake. Despite the higher total amino acid supply of cows fed red clover versus grass silage diets, further milk production responses on red clover diets were possibly compromised by an inadequate supply of methionine as evidenced by lower methionine concentration in the amino acid profile of omasal digesta and plasma. Increasing the maturity of ensiled red clover does not seem to affect silage DM intake as consistently as that of grasses. The efficiency of N utilization for milk protein synthesis was lower for red clover diets than for grass diets. It was negatively related to diet crude protein concentration similarly to grass silage diets.

Ecology of the Asian Elephant in Southern India. II. Feeding Habits and Crop Raiding Patterns

The Asian elephant's foraging strategy in its natural habitat and in cultivation was studied in southern India during 1981-83. Though elephants consumed at least 112 plant species in the study area, about 85% of their diet consisted of only 25 species from the order Malvales and the families Leguminosae, Palmae, Cyperaceae and Gramineae. Alteration between a predominantly browse diet during the dry season with a grass diet during the early wet season was related to the seasonally changing protein content of grasses. Crop raiding, which was sporadic during the dry season, gradually increased with more area being cultivated with the onset of rains. Raiding frequency reached a peak during October-December, with some villages being raided almost every night, when finger millet (Eleusine coracana) was cultivated by most farmers. The monthly frequency of raiding was related to the seasonal movement of elephant herds and to the size of the enclave. Of their total annual food requirement, adult bull elephants derived an estimated 9.3% and family herds 1.7% in quantity from cultivated land. Cultivated cereal and millet crops provided significantly more protein, calcium and sodium than the wild grasses. Ultimately, crop raiding can be thought of as an extension of the elephant's optimal foraging strategy.

Molecular phylogeny of Glyphochloa (Poaceae, Panicoideae), an endemic grass genus from the Western Ghats, India

The genus Glyphochloa (Poaceae: Panicoideae: Andropogoneae: Rottboellinae) is endemic to peninsular India and is distributed on lateritic plateaus of low and high altitude in and around Western Ghats and the Malabar Coast. The genus presumably originated and diversified in the Western Ghats. Species relationships in the genus Glyphochloa were deduced here based on molecular phylogenies inferred using nuclear ribosomal ITS sequences and plastid intergenic spacer regions (atpB-rbcL, trnT-trnL, trnL-trnF), and new observations were made of spikelet morphology, caryopsis morphology and meiotic chromosome counts. We observed two distinct clades of Glyphochloa s.l. One of these (group I') includes Ophiuros bombaiensis, and is characterized by a single-awned lower glume and a base chromosome number of 6; it grows in low elevation coastal areas. The other clade (group II') has a double-awned lower glume, a base chromosome number of 7, and is restricted to higher elevation lateritic plateaus; G. ratnagirica may belong to the group II clade, or may be a third distinct lineage in the genus. A sister-group relationship between group I and II taxa (with or without G. ratnagirica) is not well supported, although the genus is recovered as monophyletic in shortest trees inferred using ITS or concatenated plastid data. We present a key to species of Glyphochloa and make a new combination for O. bombaiensis.

Characterization and expression analysis of TNF-related apoptosis inducing ligand (TRAIL) in grass carp Ctenopharyngodon idella

TRAIL (Apo2 ligand) described as a type II transmembrame protein belonging to the TNF superfamily can induce apoptotic cell death in a variety of cell types. In the present study, a putative cDNA sequence encoding the 299 amino acids of TRAIL (GC-TRAIL) and its genomic organization were identified in grass carp Ctenopharyngodon idella. The predicted GC-TRAIL sequence showed 44 and 41% identities to chicken and human TRAILs, respectively. In a domain search, a tumor necrosis factor homology domain (THD) was identified in the C-terminal portion of TRAILs. The GC-TRAIL gene consists of five exons, with four intervening introns, spaced over approximately 4 kb of genomic sequence. Analysis of GC-TRAlL promoter region revealed the presence of a number of putative transcription factor binding sites, such as Sp1, NF-kappaB, AP-1, GATA, NFAT, HNF, STAT, P53 and IRFI sequences which are important for the expression of other TNF family members. Phylogenetic analysis placed GC-TRAIL and the putative zebrafish (Danio rerio) TRAIL obtained from searching the zebrafish database into one separate cluster near mammalian TRAIL genes, but apart from the reported zebrafish TRAIL-like protein, indicating that the GC-TRAIL is an authentic fish TRAIL. Expression analysis revealed that GC-TRAIL is expressed in many tissues, such as in gills, liver, trunk kidney, head kidney, intestine and spleen. (c) 2005 Elsevier B.V. All rights reserved.

Digital image analysis as a tool to estimate legume contributions in legume-grass swards

Summary: Productivity and forage quality of legume-grass swards are important factors for successful arable farming in both organic and conventional farming systems. For these objectives the botanical composition of the swards is of particular importance, especially, the content of legumes due to their ability to fix airborne nitrogen. As it can vary considerably within a field, a non-destructive detection method while doing other tasks would facilitate a more targeted sward management and could predict the nitrogen supply of the soil for the subsequent crop. This study was undertaken to explore the potential of digital image analysis (DIA) for a non destructive prediction of legume dry matter (DM) contribution of legume-grass mixtures. For this purpose an experiment was conducted in a greenhouse, comprising a sample size of 64 experimental swards such as pure swards of red clover (Trifolium pratense L.), white clover (Trifolium repens L.) and lucerne (Medicago sativa L.) as well as binary mixtures of each legume with perennial ryegrass (Lolium perenne L.). Growth stages ranged from tillering to heading and the proportion of legumes from 0 to 80 %. Based on digital sward images three steps were considered in order to estimate the legume contribution (% of DM): i) The development of a digital image analysis (DIA) procedure in order to estimate legume coverage (% of area). ii) The description of the relationship between legume coverage (% area) and legume contribution (% of DM) derived from digital analysis of legume coverage related to the green area in a digital image. iii) The estimation of the legume DM contribution with the findings of i) and ii). i) In order to evaluate the most suitable approach for the estimation of legume coverage by means of DIA different tools were tested. Morphological operators such as erode and dilate support the differentiation of objects of different shape by shrinking and dilating objects (Soille, 1999). When applied to digital images of legume-grass mixtures thin grass leaves were removed whereas rounder clover leaves were left. After this process legume leaves were identified by threshold segmentation. The segmentation of greyscale images turned out to be not applicable since the segmentation between legumes and bare soil failed. The advanced procedure comprising morphological operators and HSL colour information could determine bare soil areas in young and open swards very accurately. Also legume specific HSL thresholds allowed for precise estimations of legume coverage across a wide range from 11.8 - 72.4 %. Based on this legume specific DIA procedure estimated legume coverage showed good correlations with the measured values across the whole range of sward ages (R2 0.96, SE 4.7 %). A wide range of form parameters (i.e. size, breadth, rectangularity, and circularity of areas) was tested across all sward types, but none did improve prediction accuracy of legume coverage significantly. ii) Using measured reference data of legume coverage and contribution, in a first approach a common relationship based on all three legumes and sward ages of 35, 49 and 63 days was found with R2 0.90. This relationship was improved by a legume-specific approach of only 49- and 63-d old swards (R2 0.94, 0.96 and 0.97 for red clover, white clover, and lucerne, respectively) since differing structural attributes of the legume species influence the relationship between these two parameters. In a second approach biomass was included in the model in order to allow for different structures of swards of different ages. Hence, a model was developed, providing a close look on the relationship between legume coverage in binary legume-ryegrass communities and the legume contribution: At the same level of legume coverage, legume contribution decreased with increased total biomass. This phenomenon may be caused by more non-leguminous biomass covered by legume leaves at high levels of total biomass. Additionally, values of legume contribution and coverage were transformed to the logit-scale in order to avoid problems with heteroscedasticity and negative predictions. The resulting relationships between the measured legume contribution and the calculated legume contribution indicated a high model accuracy for all legume species (R2 0.93, 0.97, 0.98 with SE 4.81, 3.22, 3.07 % of DM for red clover, white clover, and lucerne swards, respectively). The validation of the model by using digital images collected over field grown swards with biomass ranges considering the scope of the model shows, that the model is able to predict legume contribution for most common legume-grass swards (Frame, 1992; Ledgard and Steele, 1992; Loges, 1998). iii) An advanced procedure for the determination of legume DM contribution by DIA is suggested, which comprises the inclusion of morphological operators and HSL colour information in the analysis of images and which applies an advanced function to predict legume DM contribution from legume coverage by considering total sward biomass. Low residuals between measured and calculated values of legume dry matter contribution were found for the separate legume species (R2 0.90, 0.94, 0.93 with SE 5.89, 4.31, 5.52 % of DM for red clover, white clover, and lucerne swards, respectively). The introduced DIA procedure provides a rapid and precise estimation of legume DM contribution for different legume species across a wide range of sward ages. Further research is needed in order to adapt the procedure to field scale, dealing with differing light effects and potentially higher swards. The integration of total biomass into the model for determining legume contribution does not necessarily reduce its applicability in practice as a combined estimation of total biomass and legume coverage by field spectroscopy (Biewer et al. 2009) and DIA, respectively, may allow for an accurate prediction of the legume contribution in legume-grass mixtures.