Online Version ISSN: 1314-412Xolume , Number

September 2012V 4 3

2012

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Genetics and Breading

Atanas Atanasov (Bulgaria)Ihsan Soysal (Turkey)Max Rothschild (USA)Stoicho Metodiev (Bulgaria)

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Nikolai Todorov (Bulgaria)Peter Surai (UK)Zervas Georgios (Greece)Ivan Varlyakov (Bulgaria)

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Marin Kabakchiev (Bulgaria)Stefan Denev (Bulgaria)Vasil Atanasov (Bulgaria)

English Editor

Yanka Ivanova (Bulgaria)

2012

Volume 4, Number 3September 2012

Online Version ISSN: 1314-412X

Effect of dietary supplementation of dried distillers grains with solubles (Zarnela) on some rumen fermentation parameters in yearling sheep

V. Radev*

Department of Morphology, Physiology and Animal Nutrition, Faculty of Agriculture, Trakia University, 6000 Stara Zagora, Bulgaria

Abstract. The investigation was carried out with five yearling sheep, Stara Zagora meat type and Pleven Blackhead crosses. The average live body weight of animals at the beginning of the experiment was 55.4 кg. The effect of dietary supplementation of dried distillers grains with solubles (DDGS) on rumen fermentation in sheep was studied. Zarnela – dried distillers grains with solubles (DDGS) is a by-product of wheat ethanol production made by the еthanol producing enterprise at Gorna Oryahovitsa, Bulgaria. The experiment consisted of two periods – control and experimental. During the control period, animals were fed a ration consisting of 1.000 kg meadow hay and 1.000 kg barley. During the experimental period, the ration contained 0.800 kg barley mash, 0.200 kg DDGS and 1.000 kg meadow hay. The following parameters were investigated: hydrogen ion concentration (рН), ammonia concentration, total volatile fatty acid (VFA) concentration.The effects of supplementing the feed of yearling sheep with DDGS were as followed:1) DDGS resulted in statistically significant reduction (p<0.01) of rumen pH before feeding. Two and a half hours after feeding, rumen pH was insignificantly higher in the experimental group. 2) The inclusion of DDGS in the animal ration during the experimental period resulted in increased total VFA concentrations (p<0.05) in the rumen before the feeding hours. The trend for higher rumen total VFA was also preserved after feeding during the experimental period. 3) After the supplementation of the diet with DDGS, rumen ammonia concentrations were significantly higher (p<0.01) in experimental animals after feeding.

Keywords: rumen fermentation, sheep digestion

Abbreviations: DDGS – dried distillers grains with solubles, VFA – volatile fatty acid

AGRICULTURAL SCIENCE AND TECHNOLOGY, VOL. 4, No 3, pp 241 - 245, 2012

performance, weight gain and meat quality in small ruminants have Introductionbeen performed.

Estrada-Angulo et al. (2007) did not observe a statistically Cellulose and crude fibres make up a major part of nutrients in significant difference in the weight gain and feed utilisation after ruminant rations. During the evolutionary development of ruminants, substitution of 15 to 35% of corn and soybean meal in yearling sheep their digestive system has evolved and improved in such a manner rations by DDSG. The addition of DDSG to the diet of yearling rams as to be able to maximally degrade and utilise rough plant feeds, at the end of the fattening period did not influence their weight (Held, which contain high percentages of cellulose and crude fibre. Under 2006). Shauer et al. (2006) suggested that DDSG mixed with cereals the influence of microbial enzymes, an intense fermentation in fore could be added to finishing lambs at a level of >22.5%. No negative stomachs of ruminants is occurring. Here, the initial degradation of impact on feed intake, growth or carcass traits was reported. carbohydrates takes place. That is why the events in fore stomachs

Despite the numerous research conducted on both national and of ruminants are essential for the supply of energy to their international scale, the effect of DDSG on digestive processes in organisms. ruminant fore stomachs and its aftereffects on digestion in general The fermentation in the fore stomachs of ruminants could be are not adequately studied. modified by dietary nutritional supplements (enzyme preparations,

The present experiment aimed at establishing the influence of probiotics, prebiotics, products from biotechnological, food and the feed supplement Zarnela on some parameters related to rumen distillery industries). The addition of these supplements aims at fermentation in yearling sheep. The results are a part of a large-scale optimising fermentation processes and achieving a more complete research on the effect of Zarnela on rumen fermentation in fore degradation of complex carbohydrates, increased rate of bacterial stomachs, fattening traits, weight gain and some blood parameters and infusorial biomass synthesis in the rumen, and increased in small ruminants.utilisation of dietary proteins. Apart from their effect on rumen

digestion, nutritional supplements influence the degradation and absorption of nutrients in intestines as well (Morgavi et al., 2001).

Material and methodsThe dried distillers grain is increasingly used as an ingredient of ruminant rations. According to Carvalho et al. (2005) dried distillers grain has high energy value, good digestibility of fat and crude fibres. To evaluate the effect of the nutritional supplement Zarnela on The nutritional supplement based on dried distillers grain with the studied parameters, a physiological trial was performed at the solubles (DDGS) combined with cereals (wheat or corn is Experimental Base of Animal Physiology Unit at the Faculty of appropriate for combination with low-quality roughages in sheep Agriculture, Trakia University, Stara Zagora. The experiment was rations, as it improves its nutritional value (Archibeque et al., 2008). conducted with five yearling sheep, Stara Zagora meat type and A number of studies on the effect of DDGS on the fattening Pleven Blackhead crosses. The average live body weight of animals

241

* e-mail: vradev@uni-sz.bg

Table 1. Chemical composition of feeds

Feed

Meadow hay

Barley mash

DDSG

88.20

89.90

85.29

9.03

9.60

26.18

28.30

5.00

6.55

1.90

1.70

2.20

1.00

1.30

3.42

DM(%)

Chemical composition (%)

Crude

protein

Crude

fibre

Crude

fat

Ash

Table 2. Daily ration fed to sheep during the control period

Feed

Meadow hay

Barley mash

Total

88.20

89.90

85.29

1.000

1.000

90.3

96.0

186.3

283

50

333

19

17

36

10

13

23

DM

intake

kg

kg

Daily intakes, g

Crude

protein

Crude

fibre

Crude

fat

Ash

90.3

77.0

52.4

219.7

Crude protein

19

14

4.4

37.4

Crude fat

10

10

6.8

26.8

Ash

Table 3. Daily ration fed to sheep during the experimental period

Feed

Meadow hay

Barley mash

DDSG

Total

0.882

0.719

0.170

1.771

1.000

0.800

0.200

283

40

13.1

336.1

DM intake kgkgDaily intakes, g

Crude fibre

242

at the beginning of the experiment was 55.4 kg. The sheep were cannula for 4 consecutive days – before feeding, 2.5 and 5 hours housed in individual pens indoors, with constant access to drinking after feeding.water and salt licks. Twenty days prior to the experiment, cannulae The following parameters were determined: hydrogen ion were surgically placed on the dorsal sac of the rumen of all animals concentration (рН), ammonia concentration, total volatile fatty acid as per Aliev (1960). The tests began after a 10-day period of (VFA) concentration.Routine methods of analysis, as described in adaptation of animals to the rations fed. previous studies of ours (Radev, 1999) were used. The results were

The experiment comprised two periods: control and statistically processed by statistical software (Statistica for experimental. During the control period, animals were fed a ration Windows, Stat. Soft. Inc.,1994).consisting of 1.000 kg meadow hay and 1.000 kg barley. During the experimental period the ration contained 0.800 kg barley mash and 1.000 kg meadow hay. Also, the experimental ration was

Results and discussionsupplemented with the feed ingredient Zarnela at 0.200 kg. The rations were offered twice daily – at 8.00 AM and 1.00 PM.

Ruminal pH is closely related to the molar ratios of volatile fatty According to manufacturer's data, Zarnela contains 90% dry acids. It is also influenced by the extent of carbohydrate and dietary matter, 33% crude protein, 7.6% crude fibre, 4.2% crude fat and organic matter fermentation in the rumen (Sutton, 1981).3.2% ash. The chemical composition of rations given throughout this

Figure 1 presents hydrogen ion concentration (рН) of ruminal experiment and of feed ingredients was obtained by proximate content in experimental animals. Before feeding, pH values were the (Weende) analysis, and was performed in the Animal Nutrition Lab at highest 6.45 (control period) and 6.14 (experimental period). After the Faculty of Agriculture, Stara Zagora. The chemical composition feeding, consequently to enhanced fermentation events in the and the amount of feed constituents of both rations are presented in rumen and VFA production, pH values decreased (p<0.05, p<0.01) Tables 1, 2 and 3.2.5 h after feed intake in both control and experimental sheep. Five Zarnela - dried distillers grains with solubles (DDGS) is a feed hours after feed intake, pH remained relatively low as compared to supplement, by-product of wheat-based ethanol production in the pre-feeding values 5.71 and 5.92 for control and experimental еthanol producing enterprise at Gorna Oryahovitsa, Bulgaria. periods, respectively. The supplementation of Zarnela resulted in According to manufacturers, Zarnela is a valuable feed ingredient ruminal pH reduction (p<0.01) before feeding.By post feeding hour appropriate for small ruminants, poultry and fish. It contains 2.5, ruminal pH in sheep fed the experimental ration was higher, but minerals, vitamins from group B (vitamin В , vitamin РР, riboflavin, 1statistically insignificant.

pantothenic acid and biotin). The content of starch is low, and it is The increased efficacy of fermentation of easily digestible

produced by carbohydrate-rich sources. The starch has fermented carbohydrates in the ration results in higher concentrations of

as a result of ethanol production. The content of amino acids, fats, volatile fatty acids in the ruminal content, and thus, on altered pH

vitamins and minerals in Zarnela is almost 3 times higher than that of values.The high pH values are beneficial for the development of

source feeds. The fibre content of Zarnela is high, and it is a rich ruminal microflora and stimulated the metabolic activity of microbial

source of energy for the animal organism. colonies, which reflects on the extent of dietary dry matter

According to the manufacturer, Zarnela could efficiently replace degradation. According to Russel and Wilson (1996), the reduction

the high-protein sunflower meal in rations, as well as a part of grain in in ruminal pH values has a considerable effect on the activity of

compound feed recipes.cellulolytic bacteria in the rumen.

Ruminal content for analysis was collected thrice daily via the

243

Figure 3. Ammonia concentration (mg/100 ml)

10.27

12.61

7.74

17.16

6.41

11.73

18

16

14

12

10

8

6

4

2

0Before feeding 2,5 h after feeding 5 h after feeding

Am

onia

mg/

100

ml

Control

Zarnela

Figure 1. pH level

6.6

6.45

6.14

5.855.93

5.71

5.92

6.4

6.2

6

5.8

5.6

5.4

5.2

Before feeding 2,5 h after feeding 5 h after feeding

pH

Control

Zarnela

Figure 2. Volatile fatty acid concentrations (mmol.l-1)

72.25

87.25

78.75

93.25 93 93.38100

90

80

70

60

50

40

30

20

10

0Before feeding 2,5 h after feeding 5 h after feeding

mm

ol/l-

1

Control

Zarnela

244

Volatile fatty acids are end products of carbohydrate digestion resulted in a marked increase in ruminal ammonia concentrations in the fore stomachs. A small amount originates from the residues of 2.5 hours after feeding but five hours after feeding, they declined deaminated amino acids when ruminants are fed protein-rich rations again. This could be due to the beneficial effect of the DSGS (Leng and . Phillipson, 1970). The great importance of VFA for the supplement on metabolic activity of ruminal proteolytic microflora ruminant organism consists in their function for energy supply and and hence on the extent of dietary protein hydrolysis. The addition of building purposes. The effect of the individual fatty acids on the Zarnela resulted in substantially increased (p<0.01) ruminal different production types is also proved.Figure 2 depicts the VFA ammonia concentrations in sheep after feeding.concentrations in the ruminal content of experimental animals.Before feeding, the total VFA content during the control

-1 period was 72.25 mmol.l . The inclusion of Zarnela in the ration Synopsis

during the experimental period resulted in increased total VFA -1content (p<0.05) in the rumen up to 87.25 mmol.l . The tendency

Zarnela - Dried Distillers Grains with Solubles from wheat, is a towards higher VFA levels with the experimental ration was

feed ingredient with relatively high carbohydrate, mineral, protein preserved in post feeding intervals too. By post feeding hour 2.5, the

and fat contents. The supplementation of small ruminant rations -1total VFA content was 78.75 mmol.l . The addition of Zarnela (yearling sheep) with Zarnela influenced some primary indices of -1resulted in post feeding VFA concentrations of 93.25 mmol.l at the fermentation in fore stomachs. The addition of Zarnela to rations

same interval. The tendency was still present five hours after altered ruminal pH, the total concentration of volatile fatty acids and -1feeding, when ruminal VFA concentrations were 93.00 mmol.l and ammonia in ruminal content of supplemented animals.Added to -193.38 mmol.l during the control and experimental periods, small ruminant rations, Zarnela altered the direction and intensity of

respectively, with insignificant differences. The time course of rumen fermentation and thus, could have an impact on digestion and ruminal VFA was compliant with the classic increase following the absorption of main feed nutrients.morning feeding. Five hours after feeding, the total VFA content in the rumen was statistically significantly higher in both tested rations (p<0.01).

The concentrations of volatile fatty acids in the fore stomachs of Conclusionruminants are influenced by multiple factors. Limiting factors are the composition of the ration, the amount of easily fermentable Zarnela resulted in statistically significant reduction (p<0.01) of carbohydrates, the feeding frequency, dietary supplementation, etc. rumen pH before feeding. Two and a half hours after feeding, rumen The so-called endogenous factors (ruminal content pH, ammonia pH was insignificantly higher in the experimental group. concentration, metabolic activity of rumen microflora, the size of The inclusion of Zarnela in the animal ration during the colonies) all have an impact on VFA content (Sutton, 1980). experimental period resulted in increased total VFA concentrations

Proteins in ruminant rations are hydrolysed by microbial (p<0.05) in the rumen before the feeding hours. The trend for higher enzymes, produced by proteolytic ruminal bacteria. The total rate of rumen total VFA was also preserved after feeding during the protein lysis in the fore stomachs is determined by several factors, experimental period. most important of which are the size and metabolic activity of the After the supplementation of the diet with Zarnela, rumen proteolytic bacterial colonies, the amount of easily degradable ammonia concentrations were significantly higher (p<0.01) in proteins in the ration, the rate of nutrients evacuation towards the experimental animals after feeding.more distant parts of the alimentary system (Kopenci and Wallase,1982). The authors suggest that non-soluble proteins are also lysed by bacteria, but in this case, the activity of ruminal

Referencesinfusoria is essential as their vitality in fore stomachs facilitates the penetration of extracellular proteolytic enzymes in feed protein

Aliev AA, 1960. A modification of the fistula method for study of particles. Dietary proteins are degraded in ruminant fore stomachs digestion in the divisions of the ruminant stomach, Sechenov at several stages. The end products of protein lysis occurring in the Journal of Physiology, Soviet union, SSSR, 46, 1505-1509 (Ru).rumen are amino acids and ammonia. The ammonia concentration Archibeque SL, Freetly HC and Ferrell CL, 2008. Feeding in the rumen is a parameter of the extent of protein degradation and distillers grains supplements to improve amino acid nutriture of absorption in fore stomachs of ruminants. The ruminal ammonia lambs consuming moderate-quality forages. Journal of Animal concentrations of sheep during both periods of the experiment are Science, 86, 691-701.presented on Figure 3. During the control period, pre-feeding Estrada-Angulo A, Contreras G, Perez A, Gamer G, Lozano O, ammonia concentration was 10.27 mg/100ml. After feeding (2.5 and Rios FG and Vazquez E, 2007. Effect of dried distillers grains 5 h) ammonia levels decreased (p<0.05) to 7.74 mg/100ml and 6.41 substituting for corn-soybean meal on growth and feed intake of mg/100ml, respectively. The reduction in ruminal ammonia Pelibuey sheep. Journal of Animal Science, 85, 572 . concentrations after feeding was most probably due to its rapid Carvalho LPF, Melo DSP, Pereira CRM, Rodrigues, MAM, absorption through the wall of the rumen and the utilisation of Cabrita ARJ and Fonseca AJM, 2005. Chemical composition, in ammonia nitrogen by protein-synthesizing bacteria.vivo digestibility, N degradability and enzymatic intestinal A different time course of ruminal ammonia concentrations was digestibility of five protein supplements. Animal Feed Science and observed with the ration supplemented with Zarnela. Before feeding,

th Technology, 119, 171–178. ruminal ammonia was 12.61 mg/100 ml. By the 2.5 hour after Held J, 2006. Feeding soy hulls and dried distillers grain with feeding, ammonia levels increased up to 17.16 mg/100 ml (p<0.01), solubles to sheep. Sauth Dakota State University, Cooperative whereas 5 hours after feeding, it was again lower (p<0.05) up to Extension Service, ExEx 2052 Access at http://agbiopubs.sdstate. 11.74 mg/100 ml. Unlike the control ration, the addition of Zarnela edu/articles/ExEx2052.pdf .

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Kopenci J and Wallase RJ, 1982. Cellular location and some Russel JB and Wilson DB, 1996. Why are cellulolytic bacteria properties of photolytic enzymes of rumen bacteria. Applied and unable to digest cellulose at low pH?. Journal of Dayry Science, Environmental Microbiology, 43, 1026 - 1033. 79,1503-1509.Morgavi DP, Beachemin KA, Sereco LV, Rode LM, McAlister TA, Schauer CS, Berg PB, Stamm M, Stecher DM, Pearson D and Iwaasa AD and Yang WZ, 2001. Resistance of feed enzymes to Drolc D, 2006. Influence of dried distillers grains of feedlot photolytic inactivation by rumen microorganisms and performance and carcass characteristics of finishing lambs. gastrointestinal proteases, Journal of Animal Science, 79, 1621- Western Dakota Sheep and Beef Day, 47,34-37.1630. Sutton JD, 1980. Digestion and end product formation in the rumen Leng RD and Phillipson AT, 1970. In: Physiology of Digestion and from product in ration. In Digestive Physiology and metabolism in Metabolism in Ruminants". Ed. by A. T. Phillipson. Newcastle upon ruminants, Eds. Y. Ruchebush and P. Thivent. MTP-press, pp. 271-Tyne. England. Oricl. Press. 290.Radev V,1999. Influence of the Yeast culture (Yea-Sacc 1026) on Sutton JD, 1981. Concentrate feeding and milk composition. In the rumen metabolism in sheep. Bulgarian Journal of Agricultural Recent advances in animal nutrition. Ed. W. Haresing. Butterworths, Science, 5, 663-668. London, 35-48.

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Development and testing of experimental sunflower hybrids obtained by using doubled haploid linesM. Drumeva

Stability of productiveness and technological qualities of diploid and triploid sugar beet varieties and hybridsG. Kikindonov

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Change of some chemical properties of alluvial-meadow soil (Mollic fluvisol) after long term fertilizationS. Todorova, N. Simeonova, K. Trendafilov, V. Valcheva

Investigation on the effect of the environment on some new common winter wheat varieties E. Penchev, K. Kostov, I. Stoeva, V. Dochev

Chemometrical analyses of Zn distribution between water and soil of dams in Chirpan Municipality, BulgariaN. Georgieva, Z. Yaneva, M. Todorova, R. Ivanova, N. Nizamov, P. Neicheva

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CONTENTS 3 / 3

AGRICULTURAL SCIENCE AND TECHNOLOGY, VOL. 4, No 3, 2012

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The manuscript should be structured as follows: Title, Names of authors and affiliation address, Abstract, List of keywords, Introduction, Material and methods,Results, Discussion, Conclusion, Acknowledgements (if any), References, Tables, Figures.The title needs to be as concise and informative about the nature of research. It should be written with small letter /bold, 14/ without any abbreviations. Names and affiliation of authorsThe names of the authors should be presented from the initials of first names followed by the family names. The complete address and name of the institution should be stated next. The affiliation of authors are designated by different signs. For the author who is going to be corresponding by the editorial board and readers, an E-mail address and telephone number should be presented as footnote on the first page. Corresponding author is indicated with *. Abstract should be not more than 350 words. It should be clearly stated what new findings have been made in the course of research. Abbreviations and references to authors are inadmissible in the summary. It should be understandable without having read the paper and should be in one paragraph. Keywords: Up to maximum of 5 keywords should be selected not repeating the title but giving the essence of study. The introduction must answer the following questions: What is known and what is new on the studied issue? What necessitated the research problem, described in the paper? What is your hypothesis and goal ?Material and methods: The objects of research, organization of experiments, chemical analyses, statistical and other methods and conditions applied for the experiments should be described in detail. A criterion of sufficient information is to be

possible for others to repeat the experi-ment in order to verify results.Results are presented in understandable tables and figures, accompanied by the statistical parameters needed for the evaluation. Data from tables and figures should not be repeated in the text.Tables should be as simple and as few as possible. Each table should have its own explanatory title and to be typed on a separate page. They should be outside the main body of the text and an indication should be given where it should be inserted.Figures should be sharp with good contrast and rendition. Graphic materials should be preferred. Photographs to be appropriate for printing. Illustrations are supplied in colour as an exception after special agreement with the editorial board and possible payment of extra costs. The figures are to be each in a single file and their location should be given within the text. Discussion: The objective of this section is to indicate the scientific significance of the study. By comparing the results and conclusions of other scientists the contribution of the study for expanding or modifying existing knowledge is pointed out clearly and convincingly to the reader.Conclusion: The most important conse- quences for the science and practice resulting from the conducted research should be summarized in a few sentences. The conclusions shouldn't be numbered and no new paragraphs be used. Contributions are the core of conclusions. References:In the text, references should be cited as follows: single author: Sandberg (2002); two authors: Andersson and Georges (2004); more than two authors: Andersson et al.(2003). When several references are cited simultaneously, they should be ranked by chronological order e.g.: (Sandberg, 2002; Andersson et al., 2003; Andersson and Georges, 2004).References are arranged alphabetically by the name of the first author. If an author is cited more than once, first his individual publications are given ranked by year, then come publications with one co-author, two co-authors, etc. The names of authors, article and journal titles in the Cyrillic or alphabet different from Latin, should be transliterated into Latin and article titles should be translated into English. The original language of articles and books translated into English is indicated in

parenthesis after the bibliographic reference (Bulgarian = Bg, Russian = Ru, Serbian = Sr, if in the Cyrillic, Mongolian = Мо, Greek = Gr, Georgian = Geor., Japanese = Jа, Chinese = Ch, Arabic = Аr, etc.)The following order in the reference list is recommended:Journal articles: Author(s) surname and initials, year. Title. Full title of the journal, volume, pages. Example:Simm G, Lewis RM, Grundy B and Dingwall WS, 2002. Responses to selection for lean growth in sheep. Animal Science, 74, 39-50Books: Author(s) surname and initials, year. Title. Edition, name of publisher, place of publication. Example: Oldenbroek JK, 1999. Genebanks and the conservation of farm animal genetic resources, Second edition. DLO Institute for Animal Science and Heal th, Netherlands.Book chapter or conference proceedings: Author(s) surname and initials, year. Title. In: Title of the book or of the proceedings followed by the editor(s), volume, pages. Name of publisher, place of publication. Example: Mauff G, Pulverer G, Operkuch W, Hummel K and Hidden C, 1995. C3-variants and diverse phenotypes of unconverted and converted C3. In: Provides of the Biological Fluids (ed. H. Peters), vol. 22, 143-165, Pergamon Press. Oxford, UK.Todorov N and Mitev J, 1995. Effect of level of feeding during dry period, and body condition score on reproductive perfor-

thmance in dairy cows,IX International Conference on Production Diseases in Farm Animals, Sept.11 – 14, Berlin, Germany, p. 302 (Abstr.).Thesis:Penkov D, 2008. Estimation of metabolic energy and true digestibility of amino acids of some feeds in experiments with muscus duck (Carina moshata, L). Thesis for DSc. Agrarian University, Plovdiv, 314 pp.

The Editorial Board of the Journal is not responsible for incorrect quotes of reference sources and the relevant violations of copyrights.

Volume 4, Number 3September 2012