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ACTA UNIVERSITATIS AGRICULTURAE ET SILVICULTURAE MENDELIANAE BRUNENSIS

Volume LXI 64 Number 3, 2013 http://dx.doi.org/10.11118/actaun201361030587

EDIBLE INSECTS – SPECIES SUITABLE FOR ENTOMOPHAGY UNDER CONDITION OF CZECH REPUBLIC

Martina Bednářová, Marie Borkovcová, Jiří Mlček, Otakar Rop, Ladislav Zeman

Received: February 19, 2013

Abstract

BEDNÁŘOVÁ MARTINA, BORKOVCOVÁ MARIE, MLČEK JIŘÍ, ROP OTAKAR, ZEMAN LADISLAV: Edible insects – species suitable for entomophagy under condition of Czech Republic. Acta Universitatis Agriculturae et Silviculturae Mendelianae Brunensis, 2013, LXI, No. 3, pp. 587–593

Since 2002, when the fi rst lecture on entomophagy took place at Mendel University in Brno, till today, participants of these educational lectures were asked to fi ll questionnaires in order to evaluate interest in entomophagy in Czech Republic and pick suitable species. Analyses of nutritional value of selected species were also performed during this time. The questionnaire was divided into several parts – suitable species, sensory properties, diffi culty of breeding and processing and respondents own attitude to the consumption of insect species. For the purpose of this study the questionnaire was evaluated using the semantic diff erential, so to create a comprehensive picture of each insect species. Based on evaluation of more than 5,000 questionnaires, certain developmental stages of seven species of insect were selected for further evaluation: Tenebrio molitor (TM) larvae, Zophobas morio (ZM) larvae, Gryllus assimillis (GA) nymphs, Locusta migratoria (LM) nymphs, Galleria mellonella (GM) larvae, Bombyx mori (BM) Pupa, Apis mellifera (AM) bee brood, while cockroaches were completely excluded for use in entomophagy. Although they are easy to breed and are available all year-round, consumers showed relatively great disgust. For all of these species, basic nutritional values were analysed, as well as content of amino acids and fattty acids. All parameters were statistically evaluated using ANOVA-1. Each species appears to be suitable for entomophagy for a diff erent reason. Generally speaking, AM, TM and GA were best accepted considering the sensory aspect, nutritional values are interesting especially in BM and GM and TM wins with simplicity of its breeding.

Tenebrio molitor, Zophobas morio, Gryllus assimillis, Locusta migratoria, Schistocerca gregaria, Galleria mellonella, Bombyx mori, bee brood, nutritional value, preference of entomophagous people

Entomophagy gains more and more interest (Premalatha et al., 2011; FAO, 2012), but also as worldwide, not only in countries with historically a functional food (Wattanathorn et al., 2012) and the long-term consumption of insects, such as Mexico possibility to contribute to sustainable life on Earth (Ramos-Elorduy, 2009B; Acuña et al., 2011), Japan (DeFoliart, 1992; DeFoliart, 1995; DeFoliart, 1997; (Nonaka, 2009), China (Chen et al., 2009) and many Ramos-Elorduy, 2009A; Yen, 2009). Europe still African countries, but also in Europe (DeFoliart, sees entomophagy as the food associated with the 1992; Oonincx and DeBoer, 2012). There the usual past of this continent. Insects are also considered food sources are suffi cient, but Brussels has already unhygienic harbors of diseases and “starvation” sounded voices to support the development of food (DeFoliart, 1999). However, in the light of new entomophagy within the European Union. At the research, when some species are examined and meeting of FAO in April 2012, strategy was created evaluated (considering their nutritional qualities to promote global consumption of insects due to the and ease of breeding) as very suitable for the diet many positives resulting from this consumption, of astronauts or residents of satellite towns in Earth not only as a source of essential nutrients orbit (Katayama et al., 2008), European perception

587 588 Martina Bednářová, Marie Borkovcová, Jiří Mlček, Otakar Rop, Ladislav Zeman

of insects is changing (Rumpold and Schlüter, ANOVA1 for each variable standard deviation and 2012). This trend can be observed also in the Tukey’s HSD test were calculated. Questionnaires Czech Republic (Borkovcová et al., 2009). Due to were evaluated using semantic diff erential. the growing interest in entomophagy in the Czech Republic the aim of this work was to (based on many RESULTS AND DISCUSSION years of experience and the latest fi ndings): suggest the most suitable species of insects for entomophagy in the Czech Republic, both in terms of nutritional Selection of insect species suitable value and acceptability to consumers, year-round for entomophagy in the Czech Republic availability in the market and diffi culty of breeding. a) Acceptable species for entomophagous people MATERIALS AND METHODS Based on the outputs of the fi rst type of Material: All insects used in this work were questionnaire, species that did not present obtained by purchasing from companies or insurmountable barrier for consumption were institutions in Czech Republic, which already have selected. Here it is necessary to mention one long experience with their breeding. Caterpillars of empirical experience of the authors, which, though BM and GM were purchased from farms at Masaryk not statistically measured yet, has infl uenced the University in Brno. Bee brood was purchased from answers of respondents, in our opinion. Although the company Přidal Brno and all other species from educational “bug banquets”, according to some the company Frýželka Brno. authors do not seem to be a very eff ective way to Preparation of insect samples for analysis: increase public interest in entomophagy (Looy Caterpillars of GM and BM were purchased alive and Wood, 2006), people who repeatedly took and stored in plastic containers without food in part in our lectures at diff erent places of Czech laboratory conditions (temperature 22 °C, humidity Republic responded to insects and dishes from 42%). In case of GM, individuals who entered the them signifi cantly more positively, willing to try post feeding phase and were leaving the containers new off ers and they considered insects completely to create pupae were gradually removed from the natural food source. containers. These individuals are emptied and From all insect species respondents chosed as reach the largest size, which is optimal state from acceptable: larvae of Tenebrio molitor (TM), Galleria the entomophagy perspective. Collected individuals mellonella caterpillars (GM), larvae of Zophobas morio were transferred to the freezer, where they were (ZM), nymphs of Locusta migratoria (LM), larvae and stored and later used for nutritional analyses and pupae of beebrood Apis mellifera (AM), nymphs cooking. BM caterpillars were kept in containers of Gryllus assimillis (GA) and pupae of Bombyx mori until they created pupae. Pupae were frozen (BM) (Fig. 1), all in order from the most to the least immediately and gradually used similarly as GM acceptable. These species are very well accepted caterpillars. Larvae of TM and ZM and nymphs of worldwide and especially BM and TM play an GA and LM were purchased alive and le two days important role in research works regarding the without food to starve. Larvae and nymphs were future of human nutrition – on Earth and in space subsequently stored in freezers. Bee brood was (Katayama et al., 2008; Hu et al., 2010; Oonincx taken from the breeder already frozen. Larvae and et al., 2010; Li et al., 2012). Special place among pupae were removed from the honeycombs using these species belongs to beebrood (AM), because tweezers before analysing. eating of it seems to be – on the contrary to other Analysis: Determination of elements’ content was mentioned species – nothing unusual (particularly performed on Varian SPECTRA AA 300. Methodics among beekeepers) in the Czech Republic. Regular of the samples’ mineralisation, conditions for consumption was confi rmed by 5% of respondents, determination of the elements and results’ and 24% of respondents tasted it at least once in their validation was stated by Soxhlet (fatty acids) and lives. Cockroaches, on the other hand, completely Kjehldal (amino acids) methods. failed in terms of acceptability, as only a thought of Statistical analysis: Respondent were at fi rst eating cockroaches caused aversion. For this reason, submitted questionnaires related to the selection cockroaches were excluded from the subsequent of appropriate species of insects, age of the analyses. respondents, education and their experience with entomophagy. For the 7 insect species with best b) Selected species in terms of diffi culty results, another questionnaire was compiled to of breeding and culinary processing evaluate subjective perception of the species, which As the toughest breedable species respondents included questions about the sensory properties selected BM, which also corresponds to reality. of selected insect species (5 questions), diffi culty of Caterpillars have to be fed fresh mulberry leaves, breeding and processing/cooking (4 questions) and which is available only at certain times of the year, the respondent’s own attitude to the consumption of and breeding is also very challenging to manual insect (3 questions). Nutritional values for 7 selected work. There are also verifi ed artifi cal diets, but they insect species were statistically evaluated using make the breeding considerably more expensive. Edible insects – species suitable for entomophagy under condition of Czech Republic 589

 PROFUSION GOOD EASY EASY EASYTO WITH WHOLE TASTY LIKEABLE CRISPY DRY HEALTHY OFTHE SMELL BREEDING HANDLING PROCESS PLEASURE BODIES  RICH  7 3,5 0  7 3,5 0  7 3,5 0  7 3,5 0

7 3,5 0

7 3,5 0  7 3,5 0

DIFFICULT MUST FOOD NOT DIFFICULT DIFFICULT GROSS STINKY SOFT JUICY TO UNHEALTHY NEVER NOTSEE FORTHE LIKEABLE BREEDING HANDLING PROCESS IT POOR

 AM ZM GA BM  LM TM GM 1: Semantic differential

On the contrary, breeding of TM seemed easy species – that is, especially in the poor countries of to respondents, clearly also because more than Africa. Very similar classifi cation was gained by GM 30% of respondents had their own experience and TM modes, the fi rst as a species, which could be with that. This species appears to be essential consumed by beekeepers and the second as – in the for entomophagy, just for simplicity of breeding opinion of respondents – “enriches” fl our routinely compared to other insect species evaluated, as during commercial processing. Many people well as other positive aspects associated with less seem anthropo-entomophagy and “a poorman’s impact on the environment compared with farm subsistence” as a synonymum (Katayama et al., 2008; animals (Oonincx and DeBoer, 2012), which is Hu et al., 2010), but there are also countries, where a big advantage when introducing as food for the consumption of insects promoted to elite dishes humans. ZM and LM were marked as relatively easy (DeFoliart, 1999; Nonaka, 2009). It depends always to breed, but only 0.2% of respondents had their on the traditions and perspectives. own experience with that. The questionnaire also Respondents would be willing to consume bee revealed that in the kitchen, when preparing insect, brood as a whole, as well as TM and GM. On the respondents would have a problem neither with the other hand, large BM pupae, larvae of ZM and LM larvae of TM and ZM, nor with LM and GM. On the nymphs would be an issue. As far as these species contrary, they would not know what to do with BM are considered, respondents would rather welcome and AM. extracts of insect or crushed insects. All respondents would also like to taste or consume AM, while BM c) Sensory evaluation of selected insect species larvae and ZM pupae attracted only a very small part Top rated were bee brood (AM) and LM. of the respondents. Surprisingly worst score was gained by the most o en consumed species – TM and ZM, but also Nutritional composition of selected insect BM, where consumers mattered above all a very species distinctive taste and consistency. Overall nutritional composition of the monitored species of insects or the contents of individual amino d) The inner attitude of respondents to acids or fatty acids did not signifi cantly diff er from selected types of insects the range published in other similar works (Tab. I– As “food for the poor” respondents evaluated III) (Bukkens, 1997; Ramos-Elorduy, 1997; Finke, mostly LM. This opinion originated probably due 2005; Mitsuhashi, 2010; Schabel, 2010; Tomotake to the common idea of the natural habitat of thie et al., 2010) and the specifi c amount of each nutrient 590 Martina Bednářová, Marie Borkovcová, Jiří Mlček, Otakar Rop, Ladislav Zeman c c a d b b b c c a b b b b 3.18 3.93 0.71 4.62 2.41 1.93 2.68 ME c c a e d b b 665.46 582.28 364.74 475.04 498.63 546.75 536.36 2.21 2.35 0.92 5.44 4.19 1.53 1.66 (kcal/100g DM) c a d b b b b 0.95 0.22 c c a 1.84 0.51 0.38 0.33 0.31 b b b b 000 000 000 000 000 000 Ash 6.91 6.42 2.65 4.25 4.26 3.84 3.65 0.01 0.01 0.02 c c a a a b b (g/100g DM) 2.94 3.55 1.33 1.15 1.29 2.38 2.23 a a a b b b b c a a b b b b 0.01 0.06 0.94 0.92 c a a e d b b 3.87 0.61 0.94 2.16 2.49 2.96 3.23 erent letters within a given nutrient and a given insect and a given nutrient lettersa given within erent 5.16 ADF 3.24 3.61 8.32 6.55 4.43 4.26 c a d b c c a a d b b insect species and given nutrient lettersa given within erent 000 0.82 0.18 0.95 insect species and a given nutrient lettersa given within erent (g/100g DM) 1.14 0.19 9.13 0.64 3.24 2.92 1.26 0.94 4.05 2.06 1.93 c a e d b b b d c a a b b b c c b b a a a 2.94 0.72 5.61 3.78 2.11 1.45 1.86 7.93 3.17 2.16 3.55 4.02 4.92 14.58 7.02 5.66 5.15 NDF 18.47 17.14 14.16 13.61 – means marked with diff with marked – means c c c a a a b (g/100g DM) e d d c a a b – means marked with diff with marked – means 1.94 2.16 1.93 0.68 0.63 0.32 1.35 – means marked with diff with marked – means a, b, c, d, c, e a, b, 5.18 2.23 2.13 3.25 st; 28.91 11.04 14.35 a, b, c, d, c, e a, b, a, b, c, d, c, e a, b, c c a a d b b c a a e b b b 3.92 3.58 1.91 2.36 7.91 3.16 2.92 c c a a b b b NFE 5.14 1.26 1.24 6.41 1.99 2.16 2.55 3.09 0.72 1.46 c a a a b b b (g/100g DM) 6.12 2.84 3.56 3.23 5.02 4.96 4.75 c a b 0 0.42 c c a b b b b 0.23 0.02 c c a e d d b 3.18 2.91 1.64 2.12 2.21 2.48 2.12 Fat 31.24 34.34 12.61 56.65 36.10 40.26 29.36 c a b c a b b b b b (g/100g DM) 000 0 00 0.08 3.12 1.88 0 1.49 0.86 0.05 0.42 2.48 0.63 1.44 1.78 1.66 1.32 1.65 a a b b b b b c a a a d b b c c a e d b b 4.52 3.64 6.58 7.23 6.24 6.93 5.91 1.12 0.18 0.29 0.49 2.13 1.03 0.98 54.38 54.25 38.41 62.21 59.23 52.62 50.86 Protein c a a b b b b (g/100g DM) c d d a a b b 3.62 1.46 1.45 2.85 3.02 2.41 2.96 2.12 12.89 20.55 c c a a d b b c c a a e d b 1.94 2.16 0.48 0.53 2.94 0.93 0.74 c a b 68.54 66.85 60.43 59.47 82.76 71.88 62.62 000 8.86 1.93 0 20.64 7.23 0.91 0.03 0.12 c a d b b b b Moisture (%) 4.57 1.95 8.64 3.93 3.52 2.91 3.12 c a a a e d b c c a d b b b c c a e e d b 1.90 2.92 1.16 5.36 4.85 3.02 6.47 4.14 3.63 3.96 (%) DM 31.56 33.28 17.33 39.67 40.61 37.45 28.22 c a b c c a d b b b 0 0.25 0 0.19 0 0.97 0 0.08 12:0 14:0 15:0 16:0 16:1 17:0 17:1 18:0 18:1 18:2 18:3 20:0 20:4 EPA DHA 1.22 0.03 0.81 ALA ASP ARG CYS GLY GLU HIS ILE LEU LYS MET PHE PRO SER THR TRY TYR VAL 3.54 4.02 2.11 4.82 2.97 3.25 3.24 P < 0.05 er at GA LM ZM TM AM BM GM GA GA LM LM ZM ZM TM TM AM AM BM BM GM GM Species Fatty acids content in selected insect species; one-way analysis of the variance-ratio test, including post-hoc Tukey’s test; test; Tukey’s including post-hoc test, the variance-ratio of insect species; analysis in selected one-way acids content Fatty Aminoacids content of selected insect species; one-way analysis of the variance-ratio test, including post-hoc Tukey’s test; test; Tukey’s including post-hoc test, the variance-ratio of insect species; analysis selected one-way of Aminoacids content Basic nutritional values of selected insect species; one-way analysis of the variance-ratio test, including post-hoc Tukey’s te Tukey’s including post-hoc test, the variance-ratio of insect species; analysis selected one-way of values Basic nutritional P < 0.05 er at P < 0.05 er at Fatty Acids (g/100g DM) (g/100g DM) Amino Acids species diff diff diff I: II: III: Edible insects – species suitable for entomophagy under condition of Czech Republic 591

in many lines meet WHO requirements for human the larvae in the dish. Most respondents also stated, nutrition (WHO, 2007). Diff erences in nutrient that AM smelled good to them. composition may be due to management practices, diff erent age of observed developmental stages and 3. Locusta migratoria nymphs especially the diet composition (Fontaneto et al., A good source of protein, the highest values 2011). of all evaluated species, low in fat. Thanks to the In terms of further processing of live insects, composition it is low caloric food source. LM has dry matter content is an important factor. As we high content of NDF and ADF, and it is a good presume, that insect distribution and production of source of Linoleic acid (18:2) and Glutamic acid. various types of food will be done in the dry state, Evaluated as a very good species with the ratio of 3:9 the cost of drying would represent a relatively high negative:positive. Manipulation was evaluated as proportion of total costs. From this perspective, little bit diffi cult. Due to their size, future consumers optimal candidate seems to be ZM with 40.6% dry would appreciate if insect would not be seen in the matter. On the other hand, processing of BM pupae, food. LM was evaluated as food for the poor (in the AM bee brood and GM caterpillars will be energy- Czech books of fi ction this species is described as consuming due to the low content of dry matter. a diet of the eremites, and this dogma persists in the The total content of fat and protein is an important Czech Republic). decision factor in a need of a food with high protein content or with high content of fat and therefore 4. Galleria mellonella caterpillars energy. Nymphs of LM had the highest protein The highest content of fat of all evaluated insect content while the total metabolizable energy was species, and the lowest content of protein. With lowest of all evaluated species. Highest content of the highest amounts of ME it is rather the energy fat (56.6% / 100 grams of dry matter) was found in component of the diet. A good source of Oleic (18:1), GM, while also this species had also the highest ME and Palmitic (16:0) fatty acids (the highest value of (665.46ME/100g of dry matter). evaluated species). This species was evaluated as the second best with the ratio of negative to the positive Nutritional and sensory profi les of selected 2:10. Negative aspects were rather diffi cult breeding species of insects and considering this species as food for the poor.

1. Bombyx mori pupae 5. Gryllus assimillis nymphs Very nutritious insect, nutrient content meets the Species with average nutritional composition requirements for human nutrition. The disadvantage in all the categories with the exception of Palmitic of this species while processed is lower dry matter (16:0) fatty acid, of which it is quite a good source. content and availability in only a very limited part It also contains high amount of Arginine (most of the year while feeding natural diet (mulberry of all evaluated species). From the perspective of leaves). BM is generally a very good source of protein entomophagous people GA is evaluated rather and fat. Semantic diff erential evaluated this species positively with the ratio of 4:8 (negative:positive). little bit negatively with ratio of negative:positive Respondents see mainly the breeding and 8:4, considering the median value 3.5, which was processing as diffi cult, some of them also did not worst of all investigated insect species. This may like the idea of biting into so bodies. This species be due to the size of pupae, and also because it is was evaluated moderately as food for the poor quite unfamiliar commodity for Czech respondents. (similarly to LM). Consistency vas evaluated as unfavourable, being rather so and juicy (respondents tend to prefer 6. Tenebrio molitor larvae hard and crunchy). Furthermore, respondents Species with average nutritional composition as see the diffi culty of breeding and processing as well. Good source of Linoleic acid (18:2) (most of a problem. The advantages seem to be easy handling evaluated species). Respondents put this species with the pupae and high nutritional value. on the fi rst place of imaginary popularity ladder, with the ratio of negative to the positive 1:11. The 2. Apis mellifera larvae and bee brood only negative point was the evaluation as food Species with high nutritional value, but like BM for the poor, which is probably associated with available only in a certain part of the year and gaining the natural occurrence of these larvae, where of this product depends on the highly professional many people noticed them – in the half-empty work of beekeepers. From all insect species AM had granaries. However, fl avour and consistency were the highest level of Glutamic acid. Respondents see evaluated as exceptionally good. Also breeding and AM as very attractive for entomophagy, the ratio of manipulation was considered easy. The species is negative and positive qualities was 4:8, which is the seen as be more or less healthy, and the majority of best ratio of all evaluated species. It is ranked as very respondents would be willing to consume foods healthy, most consumers would like to eat it again, or with high visibility of larvae (which is an advantage consume regularly and they would not mind seeing in processing, as energy-consuming mechanical 592 Martina Bednářová, Marie Borkovcová, Jiří Mlček, Otakar Rop, Ladislav Zeman

crushing or extraction of nutrients may be values were the main parts of this research. Selected eliminated). species were evaluated by consumers considering the sensory values, breeding, cooking/processing, 7. Zophobas morio larvae and personal attitude of respondents. Analyses and This species has the lowest content of moisture, questionnaires proved that, for Czech People the which could mean noticeable energy saving during following species are suitable: Tenebrio molitor (TM) further processing (especially drying). Levels of larvae, Zophobas morio (ZM) larvae, Gryllus assimillis other nutrients are rather average, with a higher (GA) nymphs, Locusta migratoria (LM) nymphs, content of Oleic (18:1) fatty acids. Respondents Galleria mellonella (GM) larvae, Bombyx mori (BM) evaluated this species rather positively with the pupae and Apis mellifera (AM) bee brood. All species ratio of 3:9, but quite an important attributes are bred in Czech Republic and there is no need were negative – the appearance was described as to import them from abroad. Apart from BM and unlovable, larvae should not rather be seen in food AM these insects are available all year round; TM and many respondents would never be willing to is easily obtainable and easy to breed. BM, GM and taste this species. On the contrary, breeding and LM can be evaluated as highly nutritive. Subjective processing was rated positively (as easy), and well evaluation by respondents lined these species as the crunch while eating larvae. The species is according to popularity as follows (from the most regarded a profusion of the rich. to the least acceptable): 1. TM, 2. GM, 3. ZM, 4. LM, 5. AM, 6. GA, 7. BM. This work proved that the interest in entomophagy in Czech Republic grows CONCLUSIONS and people do have general knowledge about usable Find suitable species of insect for entomophagy species of insects. in Czech Republic and analyse their nutritional

SUMMARY Aim of this work was to fi nd suitable species for entomophagy in Czech Republic evaluate these species considering the nutritional value and gain the attitude of Czech people to the selected species. Insects used in this work were purchased from companies or institutions in the Czech Republic, frozen and later used for nutritional analyses. Insects were also used to prepare menus served on lectures and educational events, along with several forms of questionnaires. First type of questionnaires proved that the following species are suitable for entomophagy in the Czech Republic: Tenebrio molitor (TM) larvae, Zophobas morio (ZM) larvae, Gryllus assimillis (GA) nymphs, Locusta migratoria (LM) nymphs, Galleria mellonella (GM) larvae, Bombyx mori (BM) pupae and Apis mellifera (AM) bee brood. Cockroaches were completely excluded from further analyses, as they caused aversion. Nutritional analyses of selected species supported the idea of putting these species on the menu. Most species are rather protein than fatty food, with the exception of GM larvae, which have higher content of fat than of protein. GA is an important source of Palmitic acid (16:0), GM contains a lot of Palmitic (16:0) and Oleic acid (18:1), and TM is a good source of Linoleic acid (18:2). GA was found to have signifi cantly high content of Arginine. To evaluate attitude to each species, semantic diff erential was used. This questionnaire was submitted to visitors of entomophagous events of all age categories, including pupils of all kinds of schools. Results of this survey enabled to compile a sort of “popularity ladder”. TM placed fi rst and GM second, followed by 3. ZM, 4. LM, 5. AM, 6. GA and 7. BM. The least problems with putting on the menu would cause the bee brood of AM, but this species placed fi  h considering the diffi culty of breeding. Species best accepted for entomophagy worldwide (in nutritional aspects) – pupae of BM – was evaluated as the least acceptable by Czech people. Overall, entomophagy is seen as acceptable with the possibility to include this food source in the diet of Czech people.

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Address Ing. Martina Bednářová, Department of Information Technology, University Workplaces, doc. Ing. Marie Borkovcová, Ph.D., Department of Zoology, Fisheries, Hydrobiology and Apiculture, Mendel University in Brno, Zemědělská 1, 613 00 Brno, Czech Republic, Ing. Jiří Mlček, Ph.D., Faculty of Technology, Tomas Bata University in Zlín, nám. T. G. Masaryka 5555, 760 01 Zlín, Czech Republic, doc. Ing. Otakar Rop, Ph.D., Department of Gastronomy, College of Business and Hotel Management Brno, Bosonožská 9, 625 00 Brno, Czech Republic, prof. Ing. Ladislav Zeman, CSc., Department of Animal Nutrition and Forage Production, Mendel University in Brno, Zemědělská 1, 613 00 Brno, Czech Republic, e-mail: [email protected], [email protected], mlcek@ .utb.cz, [email protected], [email protected] 594