Why are millets called coarse grains

Dissertation. for obtaining the academic degree. Doctor of Agricultural Sciences (doctor agriculturae (Dr. agr.))

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1 The ensiling of grains of large-seeded legumes as a method of preservation and the improvement of their nutritional-physiological value for monogastricians Dissertation to obtain the academic degree of Doctor of Agricultural Sciences (doctor agriculturae (Dr. agr.)) At the Professorship for Nutritional Physiology and Animal Nutrition at the Agricultural and Environmental Science Faculty of the University of Rostock Rostock, presented in 2012 by: Dipl.-Ing. agr. Annett Gefrom from Rostock born on in Berlin

2 reviewers: 1st reviewer: Prof. Dr. Annette Zeyner, Institute for Agricultural and Nutritional Sciences, Professorship for Animal Nutrition, Faculty of Natural Sciences III, Martin Luther University Halle-Wittenberg 2. Reviewer: Prof. Dr. med. vet. habil. Elmar Mohr, Professorship for Animal Health and Welfare, Faculty of Agricultural and Environmental Sciences / University of Rostock 3rd Reviewer: Prof. Dr. Hans Schenkel, State Institute for Agricultural Chemistry, University of Hohenheim 4. Reviewer: Dr. Olaf Steinhöfel, Saxon State Office for Environment, Agriculture and Geology (LfULG), Dresden, Köllitsch Date of submission: Date of defense: The promotion of the doctorate was provided by a grant from the H. Wilhelm Schaumann Foundation

3 DEDICATION I dedicate this work to my family especially in memory of my grandfather

4 The question of what can be made out of lupins was impressively demonstrated at the lupine feast of the Society for Applied Botany in Hamburg in October 1918: a lupine soup was served on a tablecloth made of lupine fibers, followed by a lupine steak fried in lupine oil and seasoned with lupine extract. There was also served: lupine margarine with 20% lupine components, cheese made from lupine protein, lupine brandy and lupine coffee. Lupine soap as well as paper and envelopes with lupine glue were also available. The soil on which field beans are grown is happy as if it had received fertilization. Pliny the Elder (23-79 AD)

5 Table of contents Table of contents SHORT VERSION ... i ABSTRACT ... iv LIST OF ABBREVIATIONS ... 53

6 Table of contents Antinutritive ingredients in ripe, store-dry field beans, peas ... Lactic acid fermentation and enzymatic conversions of necessary water Results on the effect of ensiling on the content of nutritive ingredients Results on the effect of ensiling on the content of anti-nutritional ingredients Effects of various lactic acid bacteria preparations on the possible Reduction of anti-nutritional substances in the ensiling process DISCUSSION FEED VALUE POTENTIAL OF FIELD BEANS, PEAS AND LUPINE GRAINS Nutritive feed value parameters Antinutritive feed value parameters SILICABILITY OF LARGE-SAME NATIONAL GRAINS after fermentation tests for fermentation in-vitro tests of chemical ensiling parameters. (1989) and ZIERENBERG (2000) model silages from legume grains of different types and varieties harvested with different dry matter contents Effects of ensiling on the contents of nutritive ingredients Digestibility of nutritive nutrients and energetic feed value Effects of ensiling on the contents of anti-nutritive ingredients TECHNICAL QUESTIONS ABOUT THE PROCEDURE THE SILAGE OF DAMP LEGUMINOSE GRAINS CONCLUSION SUMMARY LITERATURE LIST OF TABLES, FIGURES AND APPENDICES 10 APPENDIX Theses Thanks to CV Publications

7 Summary Summary Grain legumes are a high-quality animal feed due to their high protein and energy content. Due to the inconsistent ripening of the crops, cost-intensive technical drying of the grains is usually necessary at the conventional harvest date. The ensiling of moist legume grains could be an inexpensive and ecological preservation method for the production of protein-rich feed on conventional and ecological farms. From a technological point of view, the harvest and preservation of the still moist legume grains by lactic acid bacteria offers further advantages: o independence of the harvest time from the dry matter content o early field clearing and thus more effective use of the arable land o minimization of field losses (storage and threshing losses) o reduced costs by saving technical post-drying The aim of the present work was, in addition to investigating the suitability for silage, to examine a possible improvement in the feed value by reducing the content of anti-nutritional substances (alkaloids, oligosaccharides, phytate-phosphorus, tannins). In the event of a subsequent reduction in anti-nutritional ingredients through lactic acid fermentation, which at the same time have a protective function for the plant against pathogens, future cultivation decisions could increasingly be based on the phytosanitary situation. The investigations were carried out with ripe, store-dry field beans, peas and lupine grains of various varieties as well as with leguminous grains of various vegetation years (2005 and 2006) when harvested with a high residual moisture content (65% and 75% DM). In the starting material, the nutritional feed value parameters, selected anti-nutritional ingredients (alkaloids, oligosaccharides, phytate-phosphorus, tannins) and the chemical ensiling parameters were determined. The fermentation studies (Rostock fermentation test according to PIEPER et al and ZIERENBERG 2000 as well as the preparation of model silages with 65% and 75% DM) were carried out using various biological ensiling additives (3 repetitions per variant): o control (without ensiling additive) o molasses addition (2% of fresh mass (FM) o Lactic acid bacteria (MSB; homofermentative, Lb. plantarum; 3 * 10 5 CFU / g FM; DSM 8862, 8866) o combined addition of lactic acid bacteria + molasses (2% of FM) i

8 Summary In addition to the usual fermentation parameters, the osmolality and aerobic stability were analyzed. On the basis of the fermentation quality of the examined grain meal silages in the dry matter range of% and other results, ensiling proves to be a suitable method for preservation. The addition of lactic acid bacteria ensures successful fermentation thanks to the early and pronounced formation of lactic acid. The formation of acetic acid and alcohol was reduced. In the high DM range of 75%, lactic acid fermentation was restricted and the silage material should be re-moistened to 65% DM for safe lactic acid fermentation. A reduction of the phytate phosphorus or the alkaloid content in lupine meal silages through the ensiling process could not be proven for the time being. However, the ensiling process reduced the nutritionally flattening oligosaccharides (raffinose, stachyose, verbascose) as well as the anti-nutritional tannin and phenolic compounds in field bean and pea meal silages. From the methodical studies on the ensiling of grain meal silages with a high residual moisture content, the following essential statements can be derived from the results obtained: a. Broad beans, peas and lupine grains represent a valuable animal feed due to their high energy and protein content and its composition. B. Despite the unfavorable chemical ensiling properties, the preservation of legume grains is also possible in the high dry matter range. The preserving effect of acid formation or CO 2 atmosphere depends primarily on the residual moisture content. The residual water in the grain meal enables enzymatic conversions in the ensiling process of the substances dissolved or in suspended form in the aqueous phase. A comprehensive effect of the plant's own and microbial enzymes is possible due to the crushing of the grains and the exposure times during ensiling. c. The preservation of grain meal at approx. 75% dry matter is less based on the principle of lactic acid fermentation than on the anaerobic conditions and the high osmolalities. The preservation of the grains in this dry matter area is possible through the airtight storage. With regard to process safety, a residual moisture of 35% in the silage is recommended for lactic acid ensiling under practical conditions. d. Harvested grains can be stored without any problems, even without the use of silage aids, thanks to lactic acid or anaerobic preservation. The use of high-performance lactic acid bacteria preparations increases the safety of the fermentation process. The stake ii

9 Summary of additional sugar sources for ensiling is not necessary. Due to the high content of oligosaccharides and their breakdown in the ensiling process, legumes can be preserved by lactic acid fermentation despite their unfavorable chemical ensiling properties. e. Except for the fermentable carbohydrates, the contents of nutritional feed value parameters are not affected by the ensiling process and so the estimated high feed and energy value of legumes is retained in the grain silage. Lactic acid fermentation reduces the content of anti-nutritional ingredients (oligosaccharides, tannins) and thus improves the feed value. A reduction in the phytate phosphorus or alkaloid content through the ensiling process could not be assumed for the time being due to the irregular dynamics of the content sizes. The ensiling of legume grains harvested before maturity with a high residual moisture content can therefore be recommended as a suitable option for preservation. Taking into account the economic and work organization advantages compared to the discussed measure of chemical preservation, the form of lactic acid grain fermentation under anaerobic conditions offers an ecologically compliant processing method and is therefore also of interest for organic farming. Key Terms: Grain Legumes, Lactic Acid Wet Grain Silage, Nutritive and Anti-Nutritional Ingredients iii

10 Abstract Abstract Grain legumes are a high-quality animal feed due to their high protein and energy content. Due to the uneven ripeness of the stock at the usual harvesting time, a cost-intensive artificial drying of the grains is normally necessary. Making silages of moist legume grains could be a lower-cost and ecological conservation process for the production of protein-rich feed in conventional and organic farming. According to technical aspects, harvesting before full ripeness and the conservation of the moist legume grains through lactobacilli has further benefits: o Independence of harvest date and dry matter content. o Early field clearance and therefore a more efficient use of the land. o Minimization of nutrient loss (storage and threshing loss). o Reduced costs through elimination of artificial drying. The aim of this work was apart from the investigation of silaging, the examination of a possible improvement in the feed through the reduction of the quantity of antinutritional content (alkaloids, oligosaccharides, phytate phosphorus and tannins). The reduction of antinutritive content through the fermentation with lactobacilli, which provide the plants with protection against pathogens, future cropping decisions may be based more on the phytosanitary conditions. The experiments were performed with ripe, storage-dry field beans, peas and lupine grains of various varieties as well as with legume grains from different years (2005 and 2006) with a high remaining moisture content (65 and 75% DM). The raw material was tested for nutritive feed parameters, selected antinutrative content (alkaloids, oligosaccharides, phytate phosphorus and tannins) and the chemical fermantation parameters. The fermentation studies (Rostock Fermentation Test after PIEPER et al and ZIERENBERG 2000 and the preparation of model silage with 65 and 75% DM) followed using various biological silage additives (3 repetitions per variation): o Control (without additive) o Molasses additive ( 2% of the moist mass) o Lactic acid bacteria (LAB, homofermentative, Lb. plantarum; 3 * 10 5 CFU / g FM; DSM 8862, 8866) o Combination of LAB and molasses (2% of the moist mass) Apart from the usual fermentation parameters, the osmolality and the aerobic stability were also analyzed. Based on the fermentation quality of the sampled grain meal silage in the dry mass range% and further results, silaging is shown to be a suitable method of preservation. The addition of lactobacilli ensures the fermentation through an earlier and more comprehensive iv

11 Abstract production of lactic acid. The production of acetic acid and alcohol was reduced. Fermentation of lactic acid was limited in the high dry mass range of 75%, and the silage material should be most of the 65% dry mass to ensure fermentation with lactobacilli. A reduction of phytate phosphorus or alkaloid content in lupine silage through the fermentation process could not initially be shown. However, the nutritionally flatugen acting oligosaccharide (raffinose, stachyose, verbascose) as well as the antinutritional working tannins and phenol chains in field bean- and pea-silage were reduced. From the results of the experiments with fermented grain meal with high moisture content it is possible to draw the following conclusions: a. Field beans, peas and lupine grains are a valuable feed due to their high energy and protein content and their composition. b. Despite unfavorable chemical silage properties, the preservation of legume grains is possible even with a high amount of dry matter. The preservative effect of acid formation or CO 2 atmosphere is primarily dependent on the moisture content. The moisture found in grain meal allows enzymes to process the substances in suspension freed in the fluid phase of fermentation. A comprehensive action of plant and microbacterial enzymes is possible due to the grinding of the grains and the duration of the fermentation. c. The conservation of grain meal at approx. 75% dry matter is more due to anaerobic conditions and the high osmolality than due to fermentation with lactobacillus. The conservation of the grains in this dry matter concentration is possible due to the airtight storage. In respect of the process safety in industrial conditions, moisture content of 35% for lactic acid silage inputs is to be recommended. d. Harvest-moist grains may be stored without problems, even without the addition of silage additives, due to lactic acid or anaerobic conservation. The addition of high-performance lactobacillus preparations increases the safety of the fermentation process. The addition of sugar sources to the fermentation is not necessary. Legumes may be conserved through lactic acid fermentation, despite suboptimal chemical silage properties, due to their high content of oligosaccharides and their decomposition during fermentation. e. The content of nutritional feed value parameters are apart from the fermentable carbohydrates, not affected by the silaging process and the high feed and energy content of legumes is maintained in the grain meal silage. During the lactic acid fermentation there is a reduction of antinutritional content (oligosaccharides, tannins) and therefore an improvement of the feed quality. A reduction of the phytate-phophorus and alkaloid v

12 Abstract content during the silaging can not be assumed due to the irregular dynamic of the observed content. The fermentation of early-harvested legume grains with high moisture content can therefore be recommended as a suitable method of conservation. With respect to the economic and organizational benefits when compared to the discussed methods of chemical conservation, the lactic acid fermentation of grains under anaerobic conditions conforms to the requirements of organic agriculture and is therefore also interesting for this branch. Keywords: grain legumes, lactic acid moist grain silaging, nutritional and antinutritional content vi

13 List of abbreviations List of abbreviations AB Field bean Fig. Figure ADF Acid detergent fiber ADL Acid detergent lignin AL alcohol (Σ from ethanol, propanol, butanol, butanediol) ALK alcohol aqua dest. Distilled water AS amino acid ASM starting material aw limit water activity limit BBCH code Phenological stages of development of monocotyledonous and dicotyledonous plants, derived from: Federal Biological Research Center for Agriculture and Forestry, Federal Plant Variety Office and Chemical Industry BS Butyric acid c Concentration of osmotically active particles CCM Corn-Cob-Mix DLG Deutsche Landwirtschaftsgesellschaft DM dry matter DOS digestible organic substance DSM number German collection of microorganisms E pea E. faecium Enterococcus faecium ES acetic acid EW initial weight F dilution factor from (50 g initial weight ml distilled water) according to BLOCK & WEISSBACH (1982): = (200 ml + (50 g - (% TS * 50 g): 100)): 50 g FFS volatile fatty acids FM fresh mass GAP Common European Agricultural Policy GC-MS gas chromatography GfE Society for Nutritional Physiology GP total phenol total P total phosphorus total N / total -N total nitrogen h hour HCl hydrochloric acid HPLC High Performance Liquid Chromatography ILN Institut fü r Land use of the Faculty of Agricultural and Environmental Sciences Rostock k. A. no information CFU Colony-forming units KCl potassium chloride solution kfk germinable grains KON control conc. concentrated KT condensed tannins LAB Lactic acid bacteria L. angust. Lupinus angustifolius LD 50 mean lethal dose Lb. plantarum Lactobacillus plantarum LALLF State Office for Agriculture, Food Safety and Fisheries Mecklenburg Western Pomerania LM live mass IS loamy sand solution max. Maximum vii

14 List of abbreviations ME MJ Megajoule Metabolic energy MEL Molasses MEs Metabolic energy pork ME G Metabolic energy chicken poultry min. At least MS lactic acid MSB lactic acid bacteria mval cation exchange capacity NCIMB National Collection of Industrial and Marine Bacteria, Aberdeen, Scotland NDF Neutral detergent fiber NfE N-free extract substances (XX ) NH 3 -N ammonia nitrogen NPN non-protein nitrogen NS (LJM) precipitation (long-term mean) NSP non-starch polysaccharides NTP non-tannic phenol na not analyzed OS original substance OSM / osmol / mosmol osmolality / milliosmol Ped. acidilactici Pediococcus acidilactici P phosphorus phytate-P phytate phosphorus PK buffer capacity PS propionic acid pvk prececal digestibility PVPP polyvinylpolypyrrolidone RFO oligosaccharide (raffinose famliy of oligosaccharides); Raffinose, Stachyose, Verbascose RFT Rostocker Fermentationstest RP Pure protein SNK Student-Newman-Keuls-Test Sl clammy sand Tab. Table Tha thousand hectares TP Tannin phenol TS Dry matter TS min Minimum dry matter VDLUFA Association of German Agricultural Investigation and Research Institutes Verf KohlenVerbG Feeding ban on water World s Poultry Science Association XA crude ash XF crude fiber XL crude fat XP crude protein XS crude starch XZ raw sugar XZ MEL sugar: XZ + molasses (2% of FM) XZ RFO sugar + raffinose, stachyose, verbascose ZMP Central Market and Price Report Center GmbH Z / PK quotient from sugar and buffer capacity viii

15 1 Introduction 1 Introduction Grain legumes are used worldwide for human and animal nutrition as a protein supplier and for oil production. Legumes such as field beans, fodder peas and lupins are valuable feed due to their high protein and energy content. Since the feeding ban on animal protein feed (VerfVerbG 2001) and in the context of the controversial imports of genetically modified plants, as well as the goal of circular economy and regionalization of sustainable agricultural Production, grain legumes are of particular interest as alternative domestic protein crops for the needs-based supply of farm animals. According to the EU Commission, the proportion of the protein deficit resulting from the animal meal ban in the European Union to be substituted by plant-based crude protein carriers for the balanced supply of protein and amino acids was calculated at a required equivalent of around 2 million tonnes of animal meal for animal feed annually (ANON 2001a) . In view of the increasing global demand for crude protein in animal feed, the importance of protein supplements will continue to grow. At present, however, the protein requirements of farm animals in the European Union can only be covered by imports. For this, up to 65% of the protein carriers for use in the feed industry are sourced from the EU (EU-27), mainly in the form of soybeans (12.9 million t) and soy meal (23.6 million t) from the USA, Brazil and Argentina imported (SCHUMACHER 2006, ZINKE 2011). As a consequence of the increasing demand for inexpensive and high quality protein sources, the EU Commission (ANON 2001b) has already pointed out that the cultivation of forage crops such as native grain legumes should be promoted in Europe in order to reduce this protein deficit. For this reason, there is currently an increase in integrated research on indigenous protein carriers to meet the needs of farm animals and to secure the future of European agriculture. Although the import of soy products will continue to be decisive for the protein supply in the long term, the expansion of the area under cultivation of domestic grain legumes (currently 17% of the arable land in various EU countries; KLIEM & HEIM 2006) could increase protein supply in combination with individual, large-scale amino acids in the Essentially be stabilized. Due to their highly valued plant-building properties and positive previous crop effects, they are also of great importance in a balanced crop rotation. Legumes bind considerable amounts of atmospheric nitrogen via the symbiosis with nodule bacteria (LÜTKE ENTRUP et al. 2003). 1

16 1 Introduction This contribution to the minimization of environmental pollution (e.g. energy consumption in the production of mineral fertilizers, influence of such chemicals on the environment, CO 2 emissions) must be assigned to these crops in the economic balance. When using grain, however, the inconsistent ripening of the crops and the resulting considerable variation in the residual water content in the harvest are problematic for long-term storage. To avoid the formation of mold and the mycotoxins, which are dangerous for the health of the animals, the water content in the harvested crop must not exceed 12%. Technical drying is therefore the most common method of ensuring shelf life. Continuing rising energy prices require alternative storage methods. Lactic acid fermentation of grains with a high residual moisture content could avoid the drying costs that are usually incurred at the conventional harvest date. With the development of suitable ensiling methods for grains of the large-seeded legumes, the provision of on-farm protein-rich concentrate feed should be guaranteed without additional energy expenditure. In accordance with the ban on feeding conventional feed and synthetic amino acids in organic animal husbandry from 2012 (EG-ÖKO-VERORDNUNG 2005), the feed base can be expanded to include both organic and conventional farms. With regard to a cost-effective feed supply adapted to the work organization in year-round stable housing, the present work examined to what extent a high-quality feed can be obtained through lactic acid fermentation, also in view of the previously practiced but complex moisture preservation with organic acids, which meets the requirements corresponds to the needs and performance-based feeding of livestock and guarantees the quality of the animal products. At the same time, it was investigated whether and to what extent the levels of ingredients that reduce the feed value in grain legumes are reduced by plant and microbial enzyme effects during the ensiling process. These secondary ingredients (e.g. alkaloids, oligosaccharides, phytate phosphorus and tannins) usually have a protective function for the plant and support the plant's defense against pests and pathogenic microorganisms. The aim is therefore to make future cultivation decisions more strongly based on the phytosanitary situation (e.g. the resistance properties of certain varieties) and to establish the use of lupins, peas and field beans in the diet of farm animals, in order to partially reduce the feeding of imported soy substitute. 2

17 2 Literature Native large-grain legumes botany, cultivation and significance 2 Literature 2.1 Native large-grain legumes botany, cultivation and significance From the point of view of the botanical systematics, the legume family with approx. 600 genera and species belongs to the order of the Fabales. They are divided into 3 subfamilies, whereby the agriculturally important species are assigned to the Papilionoideae (butterflies) of different genera (ROTHMALER 1994; SCHUSTER et al. 2000). In the temperate latitudes, broad beans, peas and lupins are mainly cultivated as green manure, for green fodder or for grain production (Appendix A1 A11). Their ecological significance is based primarily on their symbiosis with nodule bacteria, which enables them to fix the nitrogen in the air. Further botanical characteristics listed in Table 1 are common to them in cultivation with correspondingly advantageous environmental effects within the crop rotation. In the case of lupins, these z. B. compared to a grain rotation with the following economic effects: Table 1: Botanical characteristics of legumes and the resulting effects in the crop rotation compared to a grain rotation of wheat (LÜTKE ENTRUP et al. 2004). The varieties permitted in Germany, their morphological properties, performance and plant cultivation requirements can be found in the Descriptive List of Varieties of the Federal Plant Variety Office (MIELKE & SCHÖBER-BUTIN 2004). Field beans (Vicia faba) For cultivation as an annual summer or winter form, the field bean needs sufficient amounts of rainfall. Due to the high need for germination water and the long vegetation period, seed placement (at a depth of approx. 4 6 cm) in deep soils at a temperature of 2 C is required from the end of February. Field beans are characterized by the 3

18 2 Literature Locals large-grain legumes botany, cultivation and importance taproots up to 1.7 m deep and their pronounced stability due to their angular, hollow stems (up to 175 cm high). One to five short-stalked flower clusters in white-reddish color combinations develop in the leaf axes. In general, variegated varieties are considered to be more resistant than white blossoming (ABEL et al. 2004). Depending on the flowering process, up to 12 pods form per plant, whereby the stages of maturity are very heterogeneous. With the highest productivity of all grain legumes, but with instability mostly caused by drought intolerance, the seed yields can fluctuate between dt / ha (KELLER et al. 1999; SCHUSTER et al. 2000; MIELKE & SCHÖBER-BUTIN 2004). Pea (Pisum sativum) As one of the oldest cultivated plants, this protein plant is still valued as food and feed. Due to the good adaptability in good site conditions with sufficient moisture, the pea spread worldwide. The crossing of the different species, subspecies and varieties resulted in a great variety of forms. The thin, deep-reaching main roots are characteristic of the peas. In grain cultivation, "leafless" or "half-leafless" varieties are increasingly being bred, in which the leaves are completely or partially transformed, as these have good stability (SCHUSTER et al. 2000). Depending on the weather, sowing should take place early in March until the beginning of April at the latest with a density of up to 90 plants per m 2 at a depth of 4 6 cm. After self-fertilization, pods with four to ten seeds each form from the inflorescences. Harvesting takes place in a combine threshing process with yields that vary depending on the location and the season between dt / ha (MIELKE & SCHÖBER-BUTIN 2004). Lupine (Lupinus ssp.) In the nomenclature, the lupine belongs to the Genisteae and finally to the genus Lupinus L. This is represented in its genetic resources with over 300 species, which originally come from two gene centers, the Mediterranean and the equatorial region of the Cordilleras (New World species) (SCHUSTER et al. 2000). From this range of species, only the white lupine (Lupinus albus L.), the yellow lupine (Lupinus luteus L.) and, due to the lower susceptibility to the anthracnose pathogen (Colletotrichum glocosporioides) and the higher yield stability and early ripeness, especially the blue lupine, have so far been included in Europe Lupine (Lupinus angustifolius L.) grown as annual cultivars (FRICK et al. 2002; ROTHMAIER et al. 2004). In addition to the varieties known as bitter lupins due to their high alkaloids content, since the breeding successes by VON SENGBUSCH in 1927, mainly low-bitter sweet lupins have been used in livestock feed (VON SENGBUSCH 4

19 2 Literature local large-grain legumes botany, cultivation and importance 1930, 1931). The growing season for lupins runs from the end of March to the beginning of September, with yields of dt / ha (white lupine), dt / ha (yellow lupine) and dt / ha (blue lupine) can be achieved (EICKMEYER 2006). Compared to peas and field beans, the blue lupine in particular has lower demands on location and rainfall and is also suitable for cultivation on sandy border locations. Cultivation and importance of native legumes In organic farming, grain legumes have been used for a long time as green manure and in field forage as native protein carriers. Due to the agricultural reform of 2001, with the related abandonment of the use of meat and bone meal in livestock feed, the market situation changed, which resulted in an increased demand for alternative vegetable protein sources and an expansion of the cultivation of grain legumes in Europe. Due to the difficult marketing conditions and unstable yields of legumes, mainly other protein crops are cultivated and so the range of legumes remains at a low level compared to other types of fruit, albeit with strong regional differentiation, or has declined in the last 10 years. In 2011, protein crops were still being grown on almost acres, mainly peas, sweet lupins and field beans, which is less than 1% of the arable land in Germany. A good half is accounted for by peas (ha), followed by sweet lupins (ha) and field beans (ha). The acreage has thus decreased by two thirds since 1998 (JOINT POSITION PAPER ON THE PROTEIN STRATEGY 2012). In 2008 and 2009, all types of grain legumes had the smallest acreage with a total of 80 tha. Field pea cultivation was at a low of 48 tha for the year 2008. Since field beans compete for better arable sites due to their drought intolerance, there was also a reduction in the area under cultivation. Lupins are mainly grown in the federal states of Mecklenburg-Western Pomerania, Brandenburg and Saxony-Anhalt. In the case of lupins, too, a clear decline in the focus of cultivation was recorded over the years, taking regional differentiations into account. In 2010, more lupine acreage was reported again for the first time (24.1 Tha). The total area under cultivation of pulses in Germany is currently showing a slight increase again (Tab. 2). The reasons for the small area under cultivation are clearly related to the contribution margin of legumes, since a positive overall balance can hardly be calculated due to high seed prices and low producer prices. 5

20 2 Literature Local large-grain legumes Botany, cultivation and importance Tab. 2: Cultivation of grain legumes in Germany from 2000 to 2012 (KLIEM & HEIM 2006, 2008, 2011; ZMP Zentrale Markt - und Preisberichtstelle GmbH; Federal Statistical Office; JOINT POSITION PAPER ZUR PROTEIN STRATEGY 2012) Type of crop in 1000 ha Broad beans 17.7 20.6 18.5 20.0 15.5 15.7 15.0 12.2 11.1 12.0 16.9 17.0 15.8 Field peas 141, 3 163.6 148.4 135.9 121.5 110.3 92.7 67.7 48.0 48.4 58.7 56.0 44.8 Lupine n. A. k. A. k. A. 45.6 35.8 38.6 33,, 3 24.1 22.0 17.8 Total 159.0 184.2 166.9 201.5 172.8 164.6 140.7 105.1 79 , 0 79.7 99.7 95.0 78.4 k. A .: no information Caused by the current legal requirements for an area payment decoupled from production (EC REGULATION No. 1782/2003), increasing economic pressure is caused in plant production. At the same time, the increasing supply of inexpensive by-products such as rapeseed meal, mash or rapeseed cake from the expansion of bioenergy production from plants in recent years reduces the incentive for additional cultivation of protein crops for fodder production. The economic disadvantage of legumes has so far not been compensated for by promoting EU agricultural policy such as the protein crop premium (55.57 / ha) (JOINT POSITION PAPER ON THE EIWEISS STRATEGY 2012). The agri-environmental programs of the second pillar according to the PLANAK resolution of February 2010 are indeed an incentive for protein crops in Germany, but are only offered by a few federal states, so that this instrument has not yet had sufficient effect (KLIEM & HEIM 2011). For decades, Germany has covered a significant proportion of its demand for protein-containing feed from imports, especially soy meal. Today around 3.0 million tonnes of rapeseed meal and 5.0 million tonnes of other oil meal, especially soy meal, are fed in Germany (JOINT POSITION PAPER ON THE EIWEISS STRATEGY 2012). Germany currently claims around 1.2 million hectares of agricultural land for protein crops outside the EU (ANON 2012). The discussions about the import of genetically modified protein carriers, yield risks and price fluctuations for soy as well as the future demands of the soy-producing countries for their own utilization in the bioenergy market underpin the argument for maximizing the level of self-sufficiency with high-quality protein through the cultivation of local plants. Grain legumes offer a complex potential for substituting soy, the cultivation of which cannot be expanded to the same extent in Europe due to climatic conditions. With a forecast increase in the world population and higher protein requirements, new areas of application will open up for field beans, peas and lupine varieties, which will improve the income of producers with increased profitability. The monetary valuation of grain legumes must therefore take into account the economic development of competing 6

21 2 Literature Local large-grain legumes botany, cultivation and importance and limited markets are assessed so that they represent a cultivation alternative with low direct costs due to the low intensity of the use of production resources, with equalization of the work peaks in cultivation and with further savings in subsequent crops due to the positive previous crop effect (Tab. 1; KLIEM & HEIM 2006). By gaining scientific information, e.g. B. through essential ingredients and the efficient use and processing with the help of new technologies, the potential of grain legumes can also be further expanded. According to KLIEM & HEIM (2006), there would be nothing to prevent an expansion of the cultivation volume from currently 17% of the arable land inside and outside Europe to%. The main overarching question is whether, after 2013, an improvement in domestic grain legumes can be achieved within the framework of the next funding period of the Common European Agricultural Policy (GAP). Possibilities for promoting domestic grain legume cultivation arise both from the current status of the discussion on the introduction of resource protection programs (greening) in order to benefit from the full direct payments of the first pillar, and from the current status of discussion on the implementation of agri-environmental programs of the second pillar in accordance with the GAP . The proposals for future agricultural policy are not expected to be adopted before 2013 (KLIEM & HEIM 2011). Associations demand that, from an agricultural policy point of view, protein crop cultivation as part of greening "should be taken into account in the CAP reform 2014 to 2020. The following measures to increase protein crop cultivation are seen as urgently necessary (JOINT POSITION PAPER ON THE PROTEIN STRATEGY 2012): o Incentives for protein crops in In the course of the reform of the Common Agricultural Policy, o long-term strengthening of agricultural research as a basis for plant breeding, o development of an overall concept for the value chain from research and breeding to cultivation and marketing or processing, o improvement of the framework for innovations in the Plant breeding 7

22 2 Literature Feed value potential of broad beans, peas and lupins when using grain 2.2 Feed value potential of field beans, peas and lupins when using grain Nutritive ingredients The value-determining feed parameters of large-seeded legumes, as listed in Table 3, largely correspond to the requirements for those relevant in animal nutrition Characteristic values ​​for the supply of essential nutrients, such as protein and amino acids, fat and fatty acids, the fiber and the energy content. Nevertheless, it must be noted that despite their similar botanical origin, they differ in the proportionate composition of the individual raw nutrients and therefore in their feed value in some cases not insignificantly. The variability of the nutrients within the species, which can be recognized according to various references, is caused by varietal differences, growth conditions, but also the selected analytical method. Tab. 3: Feed value parameters [g / kg DM] of the grains of selected legume species compared to soy extraction meal (DLG 1991, supplemented by various authors (NALLE 2009)) Dry matter Crude ash Crude protein Crude fat Crude fiber NfE Starch Sugar [g / kg FM] [g / kg TS] Soy extraction meal * 1 DLG ± 6 552 ± 24 13 ± 8 39 ± 6 329 ± 20 72 ± ± 27 Field bean DLG ± 6 299 ± 27 16 ± 6 90 ± ± ± 52 40 ± 10 Reference * ± 20 35 ± 6, 6 292 ± 39 16 ± 5.1 156 ± 20 k. A. 418 ± 4, k. A. Pea DLG ± ± 18 15 ± 5 68 ± ± ± 59 66 ± 6 Reference * ± 17 32 ± 3.3 259 ± 23 15 ± 5.2 73 ± 17 k. A. 440 ±, Blue Lupine DLG ± 6 349 ± 43 55 ± 7 159 ± ± 39 96 ± 9 54 ± 5 ​​Reference * ± 17 36 ± ± 58 60 ± ± 40 k. A. 4.2 ± 0.3 83 ± 3,, 0 4.4 k. A. Yellow Lupine DLG ± ± 33 54 ± 7 167 ± ± White Lupine DLG ± 8 376 ± 35 88 ± ± ± ± 45 71 ± 6 * 1 Soybean meal from peeled seeds, steam heated; FM: fresh mass; TS: dry matter; k. A .: no information; NfE: N-free extract substances; Field Bean * 2: THACKER (1990); BRUFAU et al. (1998); GOELEMA et al. (1999); PEREZ-MALDONADO et al. (1999); MARISCAL-LANDIN et al. (2002); HICKLING (2003); DIAZ et al (2006); PALANDER et al. (2006); Pea * 3: EASON et al. (1990); CANIBE & EGGUM (1997); GOELEMA et al. (1999); PEREZ-MALDONADO et al. (1999); ALONSO et al. (2000); MARISCAL-LANDIN et al. (2002); HICKLING (2003); WANG & DAUN (2004); DIAZ et al. (2006); PALANDER et al. (2006); NALLE (2009); NICOLOPOULOU et al. (2007); Blue lupine * 4: EASON et al. (1990); GOELEMA et al. (1999); MARISCAL-LANDIN et al. (2002); RAVINDRAN et al. (2002); GLENCROSS et al. (2003); HICKLING (2003); TORRES et al. (2005); PALANDER et al. (2006); NALLE (2009); PRIEPKE et al. (2009) Crude protein Grains of large-seeded arable legumes contain high protein contents, which increase from peas to field beans and lupins. Among the lupine species, the yellow lupine in particular is characterized by its considerable protein content. According to JÜRGENS et al. (2007), the protein yields and protein qualities between the lupine species also differ within the varieties and depending on the location. In contrast to soybeans, lupins contain 8

23 2 Literature Food value potential of broad beans, peas and lupins when using grains no trypsin inhibitors that limit the digestibility of proteins (PLARRE 1999), which increases the protein quality of sweet lupins. Overall, the biological value of pea, field bean and lupine protein can definitely be compared with that of soy (KAMPHUES et al. 2004). The storage protein fractions represented in grain legumes consist mainly of globulin (80-90%) and / or albumin (CERLETTI 1983; JEROCH 1993; ABEL 1996). Globulins are characterized by their high digestibility and, with regard to the lysine content, have a nutritionally more favorable composition of amino acids (JEROCH 1993; ABEL 1996; KNOPFE 2000; SCHUSTER et al. 2000). Amino acids The protein quality is determined by the amino acid composition and the ratio of the limiting amino acids to one another as well as their digestibility. With regard to the need for essential amino acids in monogastric animals, those in broad beans, peas and lupins, compared to cereals, have relatively high lysine contents (Tab. 4), which are nutritionally favorable (JEROCH 1993). The percentage of lysine in crude protein in lupins (4.6%) is below the order of magnitude of soy (6.0%), but it is definitely in the case of broad beans (6.2%) and peas (7.2%) comparable. Tab. 4: Crude protein and amino acid content [g / kg DM] of broad beans, peas and lupins compared to that of soy meal and wheat (DEGUSSA 2001) Lupine (sweet) broad bean extract. * Pea Wheat Soy White Yellow Blue n Raw protein essential amino acids: Methionine 2.3 2.3 1.9 2.0 2.3 2.3 7.3 cystine 4.9 5.6 4.7 3.8 3.5 3.3 8.1 lysine 15.8 17, 7 15.6 18.3 16.8 3.9 32.0 Threonine 12.0 12.6 11.1 10.1 8.8 4.1 20.7 Tryptophan 2.8 3.1 3.0 2, 6 2.2 1.7 7.0 Histidine 8.5 10.0 9.0 7.5 5.7 3.3 13.9 Isoleucine 13.4 14.9 13.1 11.7 9.5 4, 8 23.9 Leucine 23.0 25.7 22.4 21.1 16.6 9.4 40.2 Phenylalanine 13.0 14.5 12.8 12.3 11.1 6.6 26.7 Tyrosine 2 , 3 2.3 1.9 2.0 2.3 2.3 7.3 valine 13.3 14.3 12.7 13.1 10.9 6.1 25.3 arginine 34.1 41.3 35 , 9 25.7 19.9 6.8 38.9 non-essential amino acids: alanine 11.4 12.4 11.3 11.7 10.0 5.0 22.8 aspartic acid 33.6 37.3 33.2 31.0 26.8 7.4 61.0 glutamic acid 69.7 79.9 70.2 45.5 38.3 41.0 94.9 glycine 13.8 15.2 13.8 12.2 10.1 5.8 22.6 Proline 14.0 15.1 13.5 11.6 9.3 14.1 26.5 Seri n 16.4 18.2 16.4 13.5 10.9 6.6 26.8 * soy meal (peeled) 9

24 2 Literature Food value potential of broad beans, peas and lupins when using grains In addition to the sufficient levels of lysine and arginine, it should be noted, according to the evaluation of literature (DEGUSSA 2001), that the protein quality of grain legumes is due to a low tryptophan content and comparatively low amounts of sulfur-containing amino acids (methionine , Cystine) is reduced (GATEL 1994; WIRYAWAN & DINGLE 1996; WANG & DAUN 2004). Natural sources of tryptophan are only available to a limited extent and synthetic tryptophan is very expensive compared to methionine (GATEL 1994). Since, according to the usual grain / soy ration, lysine is usually the first limiting factor in the ratio of amino acids. B. for pigs the ideal amino acid ratio for an optimal utilization rate on the basis of lysine with a ratio of 0.53 0.56 (pvk methionine + cystine): 0.63 0.66 (pvk threonine) and 0.18 (pvk tryptophan ) oriented (GfE 2006). When comparing the threonine content between soy extraction meal and lupins, the relation to lysine in lupins can be assessed as favorable (ROTH-MAIER et al. 2004). According to the assessment by ABEL (1996), when comparing broad bean, pea and lupine grains, the ratio of the first limiting amino acids to one another is more balanced in lupine grains than in broad beans and peas (Tab. 5). Tab. 5: Amino acid ratio between lysine, methionine + cystine, threonine and tryptophan from grain legumes and wheat (calculated, DEGUSSA 2001) GfE 2006 * 1 lupine field bean pea wheat Sojaex. * 2 lysine = 1 white yellow blue methionine + cystine 0.53 0 .56 0.46 0.45 0.42 0.32 0.35 1.44 0.48 Threonine 0.63 0.66 0.76 0.71 0.71 0.55 0.52 1.05 0, 65 tryptophan 0.18 0.18 0.18 0.19 0.14 0.13 0.44 0.22 * 1 ratio according to prececal digestibility (GfE 2006); * 2 Soy extraction meal Especially with lupins, according to ROTH-MAIER et al. (2004) the variety-specific variability to bear, because due to the methionine content, yellow lupins provide a better supply of amino acids than white lupins and these ensure a better supply than blue lupine varieties. Low levels of methionine and cystine can limit the protein value of legume seeds as feed in terms of their substitution properties with certain feeds, but taking into account the total requirement for amino acids, the combined feeding of methionine in cereal / soy rations can counteract the excess supply. For economic and dietary reasons, the addition of legumes can supplement the lower lysine content in cereal rations and, with a sufficient amount of thioamino acids, significantly improve the protein value of both components (SCHUSTER et al. 2000). With regard to a balanced amino acid pattern and the prececal digestibility of amino acids in the ration, adequate supplementation of synthetic amino acids can avoid nitrogen surpluses and minimize nitrogen excretion (KIRCHGESSNER 2004). 10

25 2 Literature Feed value potential of broad beans, peas and lupins when using grains Fats In ripe pea and broad bean seeds, the fat content of 1 2% of DM is similarly low as in soy meal. Only in lupins, especially in white lupins, does the fat fraction achieve a noticeable level of 5 9% of the DM (Tab. 3). Thus, lupine seeds have a remarkable energy potential (WASILEWKO & BURACZEWSKA 1999; BELLOF et al. 2004). The percentage of unsaturated fatty acids in vegetable fats is particularly valuable from a nutritional point of view, whereby according to ABEL (1996) and JÜRGENS et al. (2007) the potential of legume grains for feeding is to be emphasized primarily due to the high-quality proportions of linoleic, oleic and linolenic acids with essential functions in the fatty acid composition. White lupins have a high proportion of oleic acid, followed by linoleic acid (ERBAS et al. 2005), while blue lupins contain more linoleic than oleic acid. JANSEN & JÜRGENS (2008) list the variation in the fatty acid composition of blue lupins with 19.5% saturated fatty acids, 32.4% monounsaturated and 48.1% polyunsaturated fatty acids. Carbohydrates The carbohydrate matrix of the legume grains generally consists of soluble and indigestible fractions and takes by far the largest proportion of the ingredients from% of the original substance (FRANKE 1997; PFOERTNER & FISCHER 2001; TIWARI et al. (2011); Appendix A12 A16 ). The proportionate expression of the fractional composition is subject to a strong variation between the species and varieties. With approx. 4 7% of the DM, legume seeds contain only low levels of sugar (Tab. 3). The starch contents in legume grains differ considerably, with the high starch contents in broad bean and pea grains being comparable to wheat. These legumes are therefore not only to be regarded as a protein alternative, but also to be classified as a real energy supplier in the ration. The starch content in lupins as well as in soybeans can be described as very low according to DLG information (approx. 10% of DM). According to MOHAMED & RAYAS-DUARTE (1995), WHITE et al. (2002), WRIGLEY (2003) and JANSEN et al. (2006) lupins are almost starch-free with 1 3% of DM. Non-starch polysaccharides (NSP) are therefore the main component of the carbohydrates in lupins. According to AL-KAISEY & WILKIE (1992), the function of reserve polysaccharides in lupins is taken over by galactans (galactose polymers). The NSP includes the polysaccharides (1,3 or 1,4-beta-glucans, pentosans) and other substances such as hemicelluloses and pectins, which are more easily soluble due to their binding structure, or the insoluble cellulose content and lignin (KAMPHUES et al . 2004). 11

26 2 Literature Feed value potential of broad beans, peas and lupins when using grain For the use of grain legumes in monogastric animals, their high content of pectin-like NSP, hemicelluloses and oligosaccharides must be taken into account, which has a nutritionally negative effect and lowers the feed value. Due to the lack of digestive enzymes in the organism, NSP can only be broken down by bacteria if the enzyme capacity is limited in the large intestine of the monogastric animal. Depending on their chemical composition and the solubility behavior of the NSP, absorption processes of highly digestible nutrients are impaired via the so-called cage effect or the flatogenic effect, which leads to correspondingly lower levels of convertible energy and performance depression (MURPHY 1973; STEENFELDT et al. 2003). In the case of grain legumes, the structure of the NSP is very complex compared to grain and consists mainly of a mixture of pectin-like substances in the cotyledon (CARRE et al. 1985; CHOCT 2006). In lupine kernels, these pectin substances are made up of a mixture of 1,4-beta-galactan and compounds of D-galactose, L-arabinose, L-rhamnose and galacturonic acid (CARRE et al. 1985; SMITS & ANNISON 1996). Cellulose and xylans, on the other hand, are part of the seed coat. The ratio of the individual NSP fractions differs in the comparison of the heavy legumes (PETTERSON 1998; KLUGE et al. 2002). The crude fiber content is generally quite high, but compared to field beans and peas, especially lupins, very high (Tab. 6). Tab. 6: Content of structural substances [g / kg DM] of the grains of selected legume species compared to soy extraction meal (DLG 1991, various authors) dry matter crude fiber NDF ADF ADL NSP [g / kg FM] [g / kg DM] soy extract. * 1 DLG ± 6 202 k. A. k. A broad bean reference * ± ± ± ± 20 k. A. k. A, pea reference * ± 17 73 ± ± 37 96 ± 28 k. A. k. A, 2 2, Blue Lupine Reference * ± ± ± ± 40 k. A. 350 ±, Yellow Lupine Reference * ± ± ± 7 33 ± White Lupine Reference * ± ± ± 24 36 ± ADF: Acid Detergent Fiber; ADL: acid detergent lignin; FM: fresh matter; k. A .: no information; NDF: neutral detergent fiber; NSP: non-starch polysaccharides; TS: dry matter; * 1 soy meal (peeled seeds, steam-heated): SIMON & VAHJEN (2004); STALLJOHANN (2012); * 2 broad bean: THACKER (1990); SMULIKOWSKA & CHIBOWSKA (1993); GDALA & BURACZEWSKA (1997); KNUDSEN (1997); BRUFAU et al. (1998); FLIS et al. (1999); GOELEMA et al. (1999); PEREZ-MALDONADO et al. (1999); MARISCAL-LANDIN et al. (2002); HICKLING (2003); DIAZ et al (2006); * 3 pea: EASON et al. (1990); ENGLYST & HUDSON (1996); SMITS & ANNISON (1996); CANIBE & EGGUM (1997); GDALA et al. (1997); BASTIANELLI et al. (1998); KLUGE et al. (1998); PEREZ-MALDONADO et al. (1999); KNUDSEN (1997); PERIAGO et al. (1997); GOELEMA et al. (1999); VAN BARNEVELD (1999); ALONSO et al. (2000); SMULIKOWSKA et al. (2001); MARISCAL-LANDIN et al. (2002); HICKLING (2003); WANG & DAUN (2004); ANGUITA et al. (2006); DIAZ et al. (2006); NICOLOPOULOU et al. (2007); NALLE (2009); * 4 blue lupins: EASON et al. (1990); ALLOUI et al. (1994); SMITS & ANNISON (1996); GDALA et al. (1997); GOELEMA et al. (1999); VAN BARNEVELD (1999); MARISCAL-LANDIN et al. (2002); RAVINDRAN et al. (2002); GLENCROSS et al. (2003); HICKLING (2003); NALLE (2009); PRIEPKE et al. (2009); * 5 White / Yellow Lupins: ALLOUI et al. (1994); KLUGE et al. (2002) 12

27 2 Literature Feed value potential of broad beans, peas and lupins when using grains Minerals and vitamins The mineral and vitamin contents in legume grains are similar to those of grain. The contents of iron and other minerals are high in grain legumes (TIWARI et al. 2011). Overall, the calcium and sodium levels are somewhat lower and are, in comparison, for example. In some cases below the content of soy meal, so that when using broad beans, peas and lupins it is important to ensure that the ration is adequately supplemented according to the relevant concentration norms (Tab. 7). In contrast, the grain legumes have high phosphorus and magnesium contents (JEROCH 1993).Up to 60% of the nutritionally important element phosphorus is present in the form of phytate, which is difficult for the organism to digest (GRIFFITHS & THOMAS 1981), which greatly reduces its availability, particularly in growing monogastrides. The concentration of digestible phosphorus for peas in peas is 1.9 g / kg DM (BELLOF et al. 2004). Since phytic acid also has complex bonds with e.g. B. calcium, magnesium or zinc, the addition of phytase should be standard in legume rations. The low Ca content results in a nutritionally unfavorable Ca: P ratio. A ratio of 1.5: 1 to 1: 1 is required for non-ruminants (SCHUSTER et al. 2000). The trace element contents are mostly low, apart from manganese, which is particularly abundant in white lupins (ROTH-MAIER et al. 2004). The vitamin B complex has relatively favorable levels. Overall, the vitamin content of lupins roughly corresponds to that of grain (TIWARI et al. 2011). Tab. 7: Mineral content [g / kg DM] in legume grains compared to soy meal (DLG 1973; MAKKAR et al. 1997; WASILEWKO & BURACZEWSKA 1999; PETERSEN 2002; MOSENTHIN & STEINER 2005; BÖHM 2007; STALLJOHANN & MÖLLERING 2008) (sweet) broad bean pea soy extraction white yellow blue meal * raw ash calcium 1.8 4.0 1.8 3.3 1.8 3.7 1.0 2.1 0.9 2.8 phosphorus 2.9 5, 8 4.9 9.6 2.7 6.2 4.5 7.2 3.4 5.0 7.1 Potassium ns. A k. A. Sodium k. A. k. A. k. A. 0.18 0.25 k. A. Magnesium k. A. 2.4 1.7 1.8 1.3 k. A. Soy Extraction Meal (peeled); k. A .: no information; TS: dry substance Antinutritive ingredients In addition to the feed value parameters, with regard to feeding, the anti-nutritional substances, which are usually only found in low concentrations in the grains of legumes, but which are detrimental to the use and with performance-reducing or health-endangering metabolic effects, must be taken into account. For the plants, these secondary ingredients usually have a protective function against pests (herbivores, insects, vertebrates) and pathogenic microorganisms (fungi, bacteria, viruses) or serve to attract insects, 13

28 2 Literature Food value potential of broad beans, peas and lupins when using grain for defense and UV protection (WINK 1985, 1992a; DEL PILAR VILARINO et al. 2005). Table 8 lists some of the groups of substances found in the seeds of legumes with an anti-nutritional effect. Table 8: Anti-nutritional ingredients with a performance-reducing and health-endangering effect in grain legumes (JEROCH 1993, expanded) Substance group chem. Compound Effect Occurrence Alkaloids Sparteine, Lupinine, Lupanine, Hydroxylupanine, Angustifolin Liver damage, respiratory paralysis, reduced feed intake Bitter lupins, in sweet lupins only traces Antivitamins Reduced activity of niacin broad beans Glucoside Vicin, convicin output Glucoside vicin, convicin disorder of fat metabolism, reduced fat metabolism Flatulence Disorders of the digestive processes Lupins, peas Phenol derivatives Phytic acid proteins Saponins Tannins Lectins (phytohemagglutinins) Protease inhibitors 1 MAKKAR et al. (1997); 2 FREDRIKSON et al. (2001); 3 TRUGO et al. (1993) Reduced feed intake, inhibition of proteolytic enzymes, reduced protein digestibility, impairment of the availability of minerals in monogastrides, coagulation of erythrocytes, impairment of the body's own defense mechanisms Field beans, peas, lupins Field beans, peas, lupins Field beans 1, peas, lupines Alkaloids Alkaloids are nitrogen-containing compounds and are among the most poisonous plant constituents. Due to the very effective substances, even small doses can cause special physiological effects. According to their molecular structure, the alkaloids that are most commonly found in lupins belong to the group of quinolizidine alkaloids (PETTERSON 1998; ANZFA 2001; WINK 2003). In the grains of domesticated lupins, the main alkaloids occurring in the complex alkaloid mixture listed in Table 9, whose profile is composed according to the lupine genus, are of particular interest (WINK 1992b). Lupine alkaloids are localized in all plant organs, whereby the concentration and individual variation can also depend on the corresponding plant tissues as well as seasonal or diurnal parameters (WINK & WITTE 1984; WINK 1992b; LEE et al. 2006). The individual alkaloids differ in terms of their toxicity. For Sparteine ​​z. B. a lower LD 50 found in mice than for lupanine (WINK 1994). Tab. 9: Alkaloid fractions in white, yellow and blue lupins (WINK 1992b) Lupine alkaloid fraction and proportion white: lupanine (50 80%), multiflorine (3 10%), 13-hydroxylupanine (5 15%), albine (5 15%) %) Blue: Lupanine (50 80%), Angustifolin (5 20%), 13-Hydroxylupanine (10 20%) Yellow: Lupinine (40 70%), Sparteine ​​(30 50%) 14

29 2 Literature Food value potential of broad beans, peas and lupins when using grains Alkaloids are particularly accumulated in the reproductive tissue of the seeds (WINK 1984b). Sparteine, lupine and lupanine are mainly accumulated. Bitter lupine seeds can contain increased levels of up to 4.5% of the DM (MARQUARD 2000; RÖMER 2007). Broad bean seeds and peas, on the other hand, store hardly any alkaloids (VETTER 1995; AUFHAMMER 1998). Depending on their characteristic toxicity, alkaloids damage the organism, among other things. the central nervous system and the respiratory system (WALDROUP & SMITH 1989; BIRK 1994). Current guidelines therefore prescribe a maximum total alkaloid content of 0.05% for sweet lupins used as animal feed (GARCIA 1984; JEROCH 1993; ROTH-MAIER et al. 2004). According to the ANZFA (2001), a limit value of 0.2 g alkaloid / kg is specified for lupins in the food sector. Hybridization with low-alkaloid variants allowed the alkaloid content of marketable sweet lupins to be reduced to harmless amounts of approx. 0.01% (VON SENGBUSCH 1930, 1931; ROTH-MAIER et al. 2004). This rules out intoxication. The impaired taste can, however, still have a negative impact on feed consumption, with pigs reacting very sensitively. PEARSON & CARR (1977), GODFREY et al. (1985), BELLOF & SIEGHART (1996) and ALLEN (1998) define an experimentally determined tolerance value in monogastric animals of up to 0.03% alkaloids in the feed mixture. However, much lower levels (200 mg / kg) can impair feed intake and growth in farm animals (PEARSON & CARR 1977; GODFREY et al. 1985). The alkaloid content of marketable sweet lupins is between mg / kg. Due to the environmental effect on the accumulation of alkaloids, the levels in lupine grains can vary considerably. SUJAK et al. (2006) indicate fluctuations between 0.05 and 0.24% of the TS. The aim of future plant breeding programs is a stable, sufficiently low alkaloid content in lupine varieties. On the other hand, alkaloids act as part of the plant's natural defense mechanism against microbial and herbivorous predators (WINK 1992a; BIRK 1994). If the pest pressure is high, the cultivation of bitter lupine varieties could be advantageous over the less resistant sweet lupins (WINK 2002). Oligosaccharides An essential feature of legume seeds is the high proportion of oligosaccharides, especially of the α-galactoside type. Oligosaccharides are composed of sucrose and galactose units through glycosidic linkages to form raffinose, stachyose and verbascose (TÄUFEL et al. 1962; SAINI & GLADSTONES 1986). JEROCH et al. (1999) and KIRCHGESSNER (2004) describe oligosaccharides as molecules that are made up of two to ten monosaccharide units. These water-soluble, hard-to-digest sugars with 15

30 2 Literature The potential for feed value of broad beans, peas and lupins when using grains with a low molecular weight (DEY 1985) are referred to in the literature as RFOs (raffinose family of oligosaccharides). During maturation, they are mainly accumulated in the cotyledons and less in the seed coat to protect against drought and for later embryo development (NEWTON & HILL 1983; HORBOWICZ & OBENDORF 1994). In legume seeds, the oligosaccharides are mainly raffinose, stachyose and verbascose (Tab. 10). The total content is given for field beans with around 5%, for peas between 4.5 and 7.5% and lupins with 7-15% of the DM (WISEMAN & COLE 1988; PIOTROWICZ-CIESLAK et al. 1999; MARTINEZ-VILLALUENGA et al. 2005 ). The individual oligosaccharide fractions are characteristic of the legume species, whereby WANG & DAUN (2004) and MARTINEZ-VILLALUENGA et al. (2005) point out the clear variety variability. Tab. 10: Content of sucrose and oligosaccharides [g / kg DM] in legume grains Lupine (sweet) Broad bean 3 Pea 4 Soy 3 White 1 Yellow 1 Blue 2 Sucrose 11.3 24.0 21.3 18.4 38.8 23 .6 27.0 24.8 34.0 63.8 raffinose 5.5 10.0 4.2 9.4 5.6 10.3 1.2 4.0 5.0 11.2 10.6 stachyose 49 , 7 56.0 47.6 53.8 21.2 41.8 7.4 16.0 20.0 25.3 41.3 Verbascose 6.0 14.0 22.8 25.5 11.8 23, 4 22.8 34.0 14.0 37.0 0.7 Σ α-galactoside 61.6 70.0 80.0 83.4 39.0 67.3 31.4 63.2 52.6 1. CUADRA et al. (1994); KNUDSEN (1997); KLUGE et al. (2002); 2. GDALA et al. (1997); STEENFELDT et al. (2003); PRIEPKE et al. (2009); KLUGE et al. (2002); 3. TROSZYNSKA et al. (1995); KNUDSEN (1997); 4. KNUDSEN (1997); BASTIANELLI et al. (1998); ALONSO et al. (2000); Σ α-galactosides: raffinose, stachyose, verbascose; TS: dry matter The nutritional importance lies in the flatogenic property, which has an anti-nutritional effect in monogastric animals when higher concentrations are consumed. The type of glycosidic bond of the monosaccharides is important for the enzymatic cleavage of the oligosaccharides. The anti-nutritional effect results from the lack of endogenous enzymes in the digestive system (α-1,6-galactosidase) of monogastric animals to break down the α-galactosides, so that these fractions are broken down by bacteria in the large intestine with the formation of gaseous compounds such as carbon dioxide, methane and hydrogen must (MURPHY 1973; NOWAK & STEINKRAUS 1988; BRENES et al. 1993; MINORSKI 2003). Phytate phosphorus For plants, the physiological importance of phytic acid is based on its function as a phosphorus store for the build-up of phosphatides and nucleoproteins. Phytic acid forms sparingly soluble metal compounds with bulk elements and trace elements, which are known as phytates. Phytate, the salt of phytic acid (inositol phosphoric acid), is a cyclic compound (inositol) with 6 phosphate radicals (BEIRÃO DA COSTA 1990). During grain ripening, phytate accumulates in the cotyledons (ABEL 1996). The nutritionally important element phosphorus (P) is therefore contained in legume grains in small quantities and in a form that is difficult to access for the organism. 16

31 2 Literature Food value potential of broad beans, peas and lupins when using grains The phytate content is given in broad beans as 35%, in peas as 45% and in lupins as 50% of the total phosphorus (DLG 1999; MOSENTHIN & STEINER 2005; STEINER et al. 2007). In contrast to soy, these grain legumes contain lower levels of phytate-P (PETTERSON & FAIRBROTHER 1996; Tab. 11). Tab. 11: Total phosphorus and phytate phosphorus and the proportion of phytate phosphorus in total phosphorus in various legume grains compared to wheat and soy flour Type total phosphorus Phytate phosphorus Phytate phosphorus in total phosphorus [% DM] [% ] Field bean 0.31 0.72 0.16 0, pea 0.34 0.50 0.17 0, blue lupine 0.30 0.62 0.16 0, yellow lupine 0.51 0.96 0.55 0 , White lupine 0.36 0.58 0.23 0, wheat 0.20 0.40 0.14 0, soy flour 0.57 0.77 0.40 0, PETTERSON (1998); RÖMER (1998); JEROCH et al. (1999); SELLE et al. (2003); SAUVANT et al. (2004); WANG & DAUN (2004); MOSENTHIN & STEINER (2005); STEINER et al. (2007); TS: dry matter The poor digestibility is due to the formation of the poorly soluble complexes with minerals and protein structures. Phytases can only hydrolyze dissolved phytate. Accordingly, the solubility of the phytate is the decisive factor for the availability of nutrients. Monogastrids, on the other hand, hardly have the necessary enzyme equipment in the goiter, stomach and small intestine and are v. a. relies on the hydrolytic cleavage of phytic acid-bound phosphorus by endogenous phytase activity (GRIFFITHS & THOMAS 1981; JEROCH 1993; FREDRIKSON et al. 2001). On the one hand, phytases are present in the seed itself (SILVA & TRUGO 1996; FREDRIKSON et al. 2001; GREINER et al. 2001; GREINER 2002) or are formed by bacteria (KONIETZNY & GREINER 2002; GREINER 2006). However, the activity of endogenous phytases in grain legumes in the cotylodons is only weakly developed (EECKHOUT & DE PAEPE 1994; VIVEROS et al. 2000; GREINER 2006; STEINER et al. 2007). The addition of inorganic phosphates or the supplementation of enzymes (GREINER 2006) or microbial phytase has so far been considered for a balanced phosphorus supply. Tannins Tannins belong to a special group of polyphenol polymers, which are defined as water-soluble phenols with a relatively high molecular weight between 500 and 3000 Daltons and show the typical phenolic reactions (BATE-SMITH & SWAIN 1962). Tannins exist in the plant material in a mixture with other phenolic components. From the point of view of chemical properties, there is a subdivision into hydrolyzable (split by enzymes into a sugar residue and phenol carboxylic acids) and condensed tannins (polymeric 17

32 2 Literature Food value potential of broad beans, peas and lupins when using grains Flavonoids, also called catechic tannins or proanthocyanidin, d. H. non-hydrolyzable tannins) (MAKKAR 2003; TCHONÉ 2003; HÄNSEL & STICHER 2007). The tannin fractions are among the bitter substances that reduce feed consumption in monogastric animals due to the astringent taste (JEROCH et al. 1999; FERGUSON et al. 2002). Depending on the polymerization and due to the reactive hydroxyl groups of the benzene rings, tannins form stable complex bonds with feed proteins, alkaloids and carbohydrates. The anti-nutritional effect results from the inhibition of enzymes in the digestive tract, which leads to a reduced prececal digestibility and absorption of nutrients (REDDY et al. 1985; ASQUITH & BUTLER 1986; LEINMÜLLER & MENKE 1991; JANSMAN 1993; ORTIZ et al. 1993; BHAT et al. 1985; ASQUITH & BUTLER 1986; LEINMÜLLER & MENKE 1991; JANSMAN 1993; ORTIZ et al. 1993; BHAT et al al. 1998; KHANBABAEE & VAN REE 2001; SWIECH et al. 2004). HEINZ et al. (1991) and other authors (MARQUARDT et al. 1977; SALUNKHE et al. 1990; WISEMAN et al. 1991; JANSMAN et al. 1994) attribute the anti-nutritional effect to the condensed tannins (proanthocyanidins) usually present in grain legumes. Condensed tannins are not hydrolyzed and excreted in the faeces. However, they are deposited on the intestinal wall (REED 1995) and can lead to functional disorders, whereby toxic breakdown products of the hydrolyzable tannins get into the blood (ORTIZ et al. 1994). JANSMAN et al. (1993) and MOSENTHIN et al. (1993) describe damage to the intestinal mucosa and increased endogenous protein excretion in the ileum. By absorbing high amounts of tannin, the damaged intestinal wall can also absorb condensed tannins, which can lead to damage to the internal organs if the detoxification mechanisms are overloaded (MENKE & HUSS 1987). ABEL et al. (2004) used a proportion of condensed tannins (broad beans) of 0.62% in the ration for broiler broilers and 0.37% in the TS in the feed mixes for broiler parents and noted an unfavorable feed conversion rate and increasing feed expenditure compared to the control group or significantly lower laying intensity, egg weights and hatching weights. In studies by JANSMAN et al. (1993) 0.66% condensed tannins (field bean). The results of the feeding trial showed a lower digestibility of the dry matter, the crude protein, the crude fiber and the fractions of the N-free extracts and the amino acids compared to the tannin-free ration. The different tannin fractions are manifested both species-specifically within the plant and in the legume grains, whereby they are contained in larger quantities in the seed coats of field beans and peas (Tab. 12). The seeds of multicolored flowering broad beans and pea varieties contain higher tannins (BOND 1976; BRESSANI & ELIAS 1979; GRIFFITHS 1981; CABRERA & 18

33 2 Literature Feed value potential of broad beans, peas and lupins when using grain MARTIN 1986; NOZZOLILLO et al. 1988; WISEMAN & COLE 1988; BOS & JETTEN 1989; SALUNKHE et al. 1990; VAN DER POEL et al. 1992a; DUC et al. 1995; REED 1995; VETTER 1995; RÖMER 1998; SMULIKOWSKA et al. 2001). White-flowered varieties therefore have significantly higher energy contents and a better feed value. Lupins, on the other hand, usually only contain low levels of tannin (DU PONT et al. 1994; MARISCAL-LANDÍN et al. 2002; LAMPART-SZCZAPA et al. 2003). Table 12: Tannin content [g / kg DM] in legume grains Lupine 1 broad bean 2 pea 3 soybean meal 2 white yellow blue (heated) tannins 1.7 2.6 1.7 2.1 1.2 1.9 0.04 21 , 0 2.1 6.2 2.2 1 DU PONT et al. 1994; 2 MAKKAR et al. (1997), n = 12; 3 MARISCAL-LANDIN et al; TS: dry matter vicin, convicin, lectins, protease inhibitors and saponins Leguminous grains contain other nutritionally relevant anti-nutritional ingredients.These are chemical compounds such. B. lectins, protease inhibitors and saponins. In addition to tannins, the contents of the pyrimidine glycosides vicin and convicin must be observed in the grains of field beans (HEGNAUER & HEGNAUER 2001). The anti-nutritional properties are based e.g. B. on the bitter taste (saponins), the ability of red blood cells to agglutinate, damage to the intestinal wall (lectins), protein binding and the formation of indigestible complexes (saponins), the reduced activity of proteolytic enzymes (protease inhibitors) or the hemolytic effect (saponins). Disorders of lipid metabolism are ascribed to the pyrimidine glycosides vicin and convicin (JEROCH 1993). They are split microbially in the intestinal tract and can cause hemolysis in a species-specific manner (ABEL 1996). When feeding monogastric animals with appropriate anti-nutritional ingredients in the diet, negative effects such. B. on the function of the intestinal wall cells. RUBIO et al. (1989) noted histological changes in the intestinal cell wall of the jejunum in poultry fed field beans. Changes in physiological parameters also occurred. Protease inhibitors from untreated broad bean kernels caused hypertrophy and hyperplasia in the pancreas in chicks (GERTLER et al. 1967; MARQUARDT & CAMPBELL 1973; HUISMAN & TOLMAN 1992). Due to the functional impairment of the metabolism such as reduced nutrient digestion and absorption of z. B. Amino acids showed a lower growth performance of the animals in studies (JOHNSON et al. 1986; PUSZTAI 1989; HUISMAN & TOLMAN 1992). In laying hens, significant metabolic impairments, lower egg weights and quality as well as lower hatching rates were found after increased consumption of vicin (MUDUULI et al. 1981; NABER et al. 1988; HALLE 2006). In breeding sows, these ingredients are considered to be the cause of a 19th

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