Medycyna Wet. 2010, 66 (10) 672
Praca oryginalna Original paper
The high nutritional value of yellow lupin is related
to its high protein content of high biological value and
insignificant content of non-nutritional substances. The
yellow lupin seeds include approx. 42-48% of general
protein. The main protein consists of globulins, which
constitute from 60% up to 75% of the general protein
(13). One negative property of the protein included in
the yellow lupin is its small amino-acid variety. The
content of exogenous amino-acids is high, mainly
me-thionine and cysteine, while the content of lysine and
tryptophan is low. The lupin protein is digested well
by pigs and poultry, and undergoes intensive
decom-position in the rumen. Non-nutritional substances
pre-sent in the yellow lupin seeds of high toxicity include
lupinine and sparteine as well as tannin, whose
con-tent differs depending on genetic traits of the plant
types (10, 19). There is no limit to lupin content in the
correctly composed feeding of the ruminants. Lupin
constitutes a very good high-energy and high-protein
fodder supplement for ruminants. Due to the
introduc-tion in September of 2003 of a ban on using meals
of animal origin in the feeding of ruminants (17), the
protein included in the lupin seeds may become an
alternative, filling the gap of high-protein fodder for
Influence of the addition of yellow lupin seeds
on the inhibition of methanogenesis and energetic
value of the digestive content of goats in vitro
WOJCIECH ZAWADZKI, AGNIESZKA BALCERZAK, ANDRZEJ KOTECKI*,W£ODZIMIERZ MALARZ*, ALBERT CZERSKI, STANIS£AW GRACZYK** Department of Biostructure and Animal Physiology, Faculty of Veterinary Medicine, Wroc³aw University of Environmental and Life Sciences, C. K. Norwida 31, 50-375 Wroc³aw, Poland
*Department of Crop Production, Faculty of Life Sciences and Technology,
Wroc³aw University of Environmental and Life Sciences, Pl. Grunwaldzki 24A, 50-363 Wroc³aw, Poland **Department of Immunology, Patophysiology and Veterinary Prevention, C. K. Norwida 31, 50-375 Wroc³aw
Zawadzki W., Balcerzak A., Kotecki A., Malarz W., Czerski A., Graczyk S.
Influence of the addition of yellow lupin seeds on the inhibition of methanogenesis and energetic value
of the digestive content of goats in vitro
Summary
The goal of the research has been to determine the influence of ground seeds of yellow lupin of Markiz and Teo types on the value of selected fermentation parameters, such as pH, methane production and energy value of goats rumen content in vitro. The samples for analysis were drawn at the 1, 2, 4, 6 and 24 hours of fermentation. Fermentation was conducted in specially designed vessels. Based on conducted tests, it was determined that the addition of yellow lupin to the fermented rumen contents, especially in the amount of 5 g per vessel, caused the inhibition of methanogenesis (both Markiz type and Teo type) in the 4th hour by 50%
(p < 0.001) and decrease of energy level by 45%.
During bacterial fermentation, gasses are also produced in the rumen. The most important ones include carbon dioxide and methane. The quantitative proportion of these two gases (CO2 : CH4) is frequently used for the evaluation of the fermentation process correctness. It increases with the introduction of nutritive fodder, and decreases when feeding with bulky feed. The production of large amounts of methane during fermenta-tion causes considerable energy losses. The losses can be reduced through a modificafermenta-tion of the nutrifermenta-tional dose or introduction of methanogenesis inhibitors into the fodder. The addition of lupin of both Markiz and Teo type caused the reduction of methane production. This is very important from the point of view of animal production profitability, because it results in the better use of fodder by the animal. The reduction of methane production by the addition of lupin to the nutritional dose is also of ecological importance. Methane is classi-fied as a greenhouse gas, and its emission reduction through the ruminants diet modification has been the subject of numerous publications.
Medycyna Wet. 2010, 66 (10) 673
animals. Intensively growing and intensively used
ani-mals (e.g. dairy cows) show a particularly high demand
for high-protein feed.
The preliminary observation that faba bean and
skullcap root act (12, 23) similarly in methanogenesis
inhibition (2-5, 9, 14, 15, 20, 21) suggested the idea
of defining the levels of energetic value, methane
pro-duction, total protein and volatile fatty acids in rumen
contents of goats after giving to a sample rumen
yellow lupin. Although the influence of mentioned kind
of plants on the course rumen fermentation both in vivo
and in vitro was the subject (5, 11) of our earlier
expe-riments, until now the mechanism of these additives
on ruminant feeding was not explained decisively. In
this work two types of yellow lupin: Markiz and Teo,
were compared.
The aim of the present article was therefore to
determine the main role of yellow lupin seeds added
to rumen contents of goats as a product characterized
by three chosen parameters of fermentation taking
place in the forestomach: pH, energetical value and
methane production.
Material and methods
The examination was conducted on 8 goats, interracial hybrids (at the age from 2 to 4 and with a body mass from 40 up to 45 kg), from which the rumen fluid and contents were taken 2.5 hours after morning feeding. Goats had been fed with hay (50%) and concentrates (50%) according to feeding standards. The daily dose of fodder amounted to 1.8-2.3 kg and was given to goats twice a day in equal doses at 7:30 a.m. and 2:30 p.m. The material for analysis was collected through the cannule of the dorsal rumen sack, made in accordance with Dejneka and Ziêba method (6). Incubation of the collected contents was conducted in the vessels (figure 1), made in accordance with the Barnett and Reid method (1) according to Zawadzkis modification (22). The incubation environment in the vessels included 50 ml of the collected rumen fluid contents, 50 ml of distilled water and 50 ml of McDougalls artificial saliva of pH = 6.9 with the following composition (in 1 dm3 of distilled water): NaHCO
3
9.60 g, KCl 0.60 g, CaCl2 0.04 g, Na2HPO4·12 H2O 9.15 g, NaCl 0.45 g, MgSO4·7 H2O 0.11 g, ZnSO4·H2O 0.06 g, CaCl2·6 H2O 0.01 g, NH4HCO3 0.5 g. (1, 15). The control sample consisted of the vessels including 50 ml of the collected contents, 50 ml of distilled water and 50 ml of artificial saliva. Other vessels included 50 ml of the fil-tered rumen contents, 50 ml of distilled water and 50 ml of artificial saliva as well as ground seeds composed of the yellow lupin of Markiz and Teo types in three different do-ses: 1 g, 2 g and 5 g per vessel. The total incubation volume of the sample in the vessels was 150 ml (with total vessel volume of 250 ml). The vessel leaktightness was secured by means of polished plugs and valves, covered with sealing silicon lubricant. Nitrogen was added to vessels under pres-sure to obtain anaerobic conditions. The gas was forced through the vessel content for 10-15 minutes, after which all valves were closed. The maintaining of anaerobic conditions was checked with a DO-5508 device (manufactured by
Lutron). The vessels prepared in this way were then placed in the seats of a shaker with a water bath of 39-42°C tem-perature and an amplitude of 4 cycles per minute (20). The samples were collected in the 1, 2, 4, 6 and 24 hours of incubation in order to determine the level of total energy released in the sample, and the methane as well as pH of the fermented rumen contents. Total energy in rumen samples was determined according to Gawêcki and Jeszka (8) after preliminary preparations in a vacuum evaporator or dryer. Methane concentration was measured by means of a methano-meter of Barbara-3 type (20, 22) and the method described by Varadyova (18). The pH reaction of the collected sam-ples of the incubated rumen contents was measured with a pH-meter (pX-processor PM-600).
Statistical analysis. Research results were elaborated statistically using the t-Student test and ANOVA variation analysis.
Results and discussion
The results obtained in the studies are presented in
tables 1 and 2. The addition of lupin to the fermented
rumen contents resulted in the increase of energy
rele-ased levels, particularly of the 5 g dose of Markiz type
in the 2
ndhour (63.6%), 1
sthour (53.3%) and in the 4
thhour (44.4%) of fermentation. Smaller doses of lupin
seeds of Markiz type brought about significant
chan-ges of the energy levels (from 8.3% to 23.7%) in the
fermented rumen contents in comparison to the blind
test (tab. 1). During bacterial fermentation gasses were
also produced in the rumen. The most important ones
include: CO
2and CH
4(17, 24, 25). The quantitative
proportion of the above-mentioned gasses is very often
used for the evaluation of the fermentation process
correctness (4, 20, 21). It increases the introduction
of nutritive fodder and decreases when feeding with
bulky feed (20-22). The production of large amounts
of methane during fermentation causes energy losses
(10, 20-22).
The methane changes of the fermented rumen
con-tents are shown in table 2. A significant decrease was
observed of the contents with the addition of Markiz
as well as Teo type seeds at the dose of 5 g in the 1
sthour and the 4
thand 6
thhours (p < 0.01 and p < 0.001).
The reduction of methane production by lupin seeds,
Fig. 1. Vessel for incubation of the goat rumen contents sam-ples with the supplement of yellow lupin grainMedycyna Wet. 2010, 66 (10) 674
in addition to the nutritional doses, is very important
in saving the energy losses and for the better use of
fodder by the animals (7, 16).
The pH changes of the rumen contents were
signifi-cant only in the 4
thhour in addition to Markiz as well as
Teo type seeds in the dose of 5 g per vessel (p < 0.01).
Lupin seeds may become a nutritional alternative
which will substitute animal-origin fodder withdrawn
from ruminant feeding. The addition of both types of
the yellow lupin seeds to the fermented rumen content
samples also reduced the methanogenesis, thus
con-tributing indirectly to the reduction of greenhouse gas
emission.
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Adres autora: prof. dr hab. Wojciech Zawadzki, ul. Norwida 31, 50-375 Wroc³aw; e-mail: wojciech.zawadzki@up.wroc.pl
Explanations: Essential statistical changes of methane concentration in comparison to methane concentration in the control group; *** p < 0.001, ** p <0.01, * p <0.05 Tab. 1. Mean level of energy released in the sample in rumen of goats (kcal · g D. M1) during incubation in in vitro research with the addition of 1 g, 2 g, and 5 g of Markiz and Teo lupin seeds
e m it n o it a b u c n I Cgornourtpol MARKIZ TEO g 1 2g 5g 1g 2g 5g r u o h 1 % e s a e r c n i y g r e n E 3.0 133..43 136.5.7 45.36*.3* 2.4 2.8 2.8 r u o h 2 % e s a e r c n i y g r e n E 3.3 132..72 138..92* 56.43*.6** 33..11 3.2 3.3 r u o h 4 % e s a e r c n i y g r e n E 3.6 38..93 242.4.2* 54.24*.4** 3.4 3.5 38..93 r u o h 6 % e s a e r c n i y g r e n E 3.8 143..32 42.37.*7* 53.14*.*2* 3.6 3.7 47..91 r u o h 4 2 % e s a e r c n i y g r e n E 4.4 *44..56* 153.0.6 155..19 3.9 4.1 44..65 Explanations: Essential statistical changes of energy level in comparison to energy in the control group; *** p < 0.001, ** p < 0.01, * p < 0.05
Tab. 2. Mean level of methane production [% vol.] during incubation of rumen fluid in goats in in vitro research with the addition of 1 g, 2 g and 5 g of Markiz and Teo lupin seeds
e m it n o it a b u c n I Conrtol p u o r g Z I K R A M TEO s r u o h 1g 2g 5g 1g 2g 5g 1 1.40 0.50** 0.48** 0.45*** 0.52*** *0,40*** *0.49*** 2 1.30 0.80** 0.95** 0.85*** 0.82*** 0.78** 0.86** 4 1.22 0.72** 0.70** 0.65*** 1.05*** 0.72** 0.80** 6 1.25 0.71** 0.73** 0.63*** 0.90*** 0.95** 0.72** 4 2 1.35 *0.65*** *0.65*** 0.97*** 0.92*** 0.92** 0.77**
