Supplementation of DFM in ammoniated palm fronds on fermentability and bacteria population
Methane gas has a very significant contribution to the increase in greenhouse gases (GHG) globally. The livestock sector, especially ruminants, causes the issue of increasing GHG concentrations. The chapter presents the issue of reducing methane gas production from cattle. Various experiments to reduce methane gas production from ruminants have been carried out and have shown varying results. This series of results of the author\'s research on reducing methane gas production in livestock in beef cattle based on agriculture by-product to animal feed is addressed with this background. Agriculture by-products such as oil palm fronds and rice straw can be used to feed beef cattle in Indonesia. However, agriculture by-product as animal feed can reduce feed efficiency and increase methane gas production due to the high lignin content. Therefore, various alternatives are carried out to optimize the utilization of this plantation waste. One of them is the use of feed additives and methanogenesis inhibitors. The author\'s series of research using feed additives (direct-fed microbial) and various methanogenesis inhibitors (plant bioactive compounds and dietary lipids) were tested to determine their effect on nutrient digestibility and methane gas production in feed based on plantation waste. Experiments were carried out in vitro and in vivo on various types of ruminants. Plant bioactive compounds such as tannins are proven to reduce methane production through their ability to defaunate in the rumen. Tannins may also have direct effect on methanogens and indirectly by reducing fiber digestion. In addition, direct-fed microbial (DFM) feed additives such as Saccharomyces cerevisiae, Bacillus amyloliquifaciens, and Aspergillus oryzae can be used in ruminants to increase livestock productivity. Furthermore, virgin coconut oil as a dietary lipid contains medium-chain fatty acids, mainly lauric acid, which can inhibit the development of ciliates of protozoa and methanogenic bacteria that produce methane in the rumen.
- feed additive
- direct fed microbials
- virgin coconut oils
- tannins and saponin
- methane emissions
- beef cattle
- ammoniated palm frond
The main problem in the development of ruminant livestock production in Indonesia, such as beef cattle, is the difficulty of meeting the availability of forage sustainably, both in quality and quantity. Therefore, the use of plantation waste such as palm fronds, rice straw as animal feed is an alternative that can be done to overcome the problem of feed availability. The utilization of plantation waste as ruminant feed is still minimal due to the high content of lignin  which causes low digestibility [1, 2, 3]. To optimize plantation waste as animal feed, it is necessary to combine processing techniques and optimize bioprocesses in the rumen , which aims to increase the microbial population and streamline the fermentation process in the rumen.
Supplementation of direct-fed microbial (DFM) and methanogenesis inhibitors is a way that can be done to increase the efficiency of rumen fermentation [3, 4, 5]. DFM is a feed additive product that contains a source of live microorganisms , can modify the rumen ecosystem , synthesize nutrients so that their availability can increase livestock growth .
High-fiber feeds such as plantation waste reduce not only the efficiency of feed use  but also increase the production of methane gas (CH4) . In the livestock sector, methane is one of the gaseous products of fermented feed ingredients by rumen microbes. Ruminants account for more than 75% of methane emissions from total greenhouse emissions . The release of methane causes an increase in the concentration of CH4 in the air and causes energy loss of 6–13% from the feed . Many livestock nutritionists try to reduce methane production because they feel responsible for the contribution of the livestock sector to atmospheric pollution by methane, as one of the pollutants that is always associated with global warming . Decreased methane production in the rumen is closely related to the metabolic activity of protozoa . Ciliated protozoa in the rumen are in symbiosis with methane bacteria, so that by reducing the population of ciliated protozoa, it will reduce the availability of hydrogen for the formation of methane .
Tannins are plant bioactive compounds that can reduce methane production because they act as protozoal defaunation agents . The results of the meta-analysis of
Based on the description above, this chapter book presents several reviews of the results of the author’s research, which combines a combination of processing techniques and optimization of bioprocesses in the rumen to increase the value of benefits from plantation waste that can be packaged into complete quality rations, able to increase livestock productivity and reduce beef cattle methane production.
2. Direct fed microbial and virgin coconut oils on methane gas production
2.1 Effect direct-fed microbes on rumen microbial population
Direct-fed microbes (DFM) have comparable results to probiotics. DFM is a feed product that contains a source of live microorganisms . DFM is commonly used as a supplement to increase livestock production. DFM commonly used in ruminants is yeast. DFM works to modify the rumen ecosystem to create an optimal environment for the development of rumen microbes. The provision of DFM as an additive to live microbes in the feed will affect the host by improving the balance of rumen microorganisms .
The three-stage series of research has been conducted by Suryani
|VFA (mM)||NH3 (mM)||Bacteria population (cell mL−1)|
|P0||108.35e||12.28d||1.61 x 109e|
|P1||130.69ab||14.97ab||2.49 x 109ab|
|P2||125.10cd||14.47ab||2.37 x 109bc|
|P3||123.24cd||13.73bc||2.40 x 109bc|
|P4||126.97bc||15.25a||2.41 x 109bc|
|P5||132.55a||15.75a||2.55 x 109a|
|P6||121.38d||13.06cd||2.35 x 109c|
|P7||121.38d||12.78cd||1.93 x 109c|
The results showed that DFM supplementation in feed based on plantation waste in the form of ammoniated palm frond could increase rumen fermentability. The bacterial population increased from 1.61 x 109 to 2.35 x 109 cell mL-1. These results are following the results of research [1, 9] where the addition of probiotics in the ration can stimulate the development of microbes in the rumen and increase the digestibility of food in livestock. The way yeast works in the rumen can utilize oxygen to ensure anaerobic conditions for rumen bacteria and stimulate specific rumen bacterial populations  (Figure 1). However, there was a tendency for the bacterial population to decrease in the combination supplementation of three types of DFM (P7). It was suspected that there was an accumulation of rumen microbial growth so that bacteria in the rumen competed in digesting feed. The total NH3 and VFA concentrations increased from 12.28 mM to 14.28 mM and 108.35 mM to 125.90 mM. Desnoyers
Furthermore, DFM fungal
Yeast culture uses oxygen to metabolize feed particles into sugars and oligosaccharides to produce peptides and amino acids as end products used by bacteria. Most rumen microorganisms are anaerobic, so the utilization of oxygen by yeast culture will increase the optimum conditions in the rumen. These conditions will protect the anaerobic rumen bacteria from damage by O2. They created better conditions for the growth of cellulolytic bacteria so that the number of cellulolytic bacteria increases and improves digestion in the rumen .
Yeast activity as DFM can regulate rumen biological activity by stimulating lactic acid utilization and reducing ammonia production, so that rumen pH is stable and increases nutrient absorption and VFA profile  . Supplementation can support livestock productivity by increasing intestinal development, mucosal immunity, nutrient absorption, and inhibiting pathogenic bacteria. This will have an impact on improving livestock health and performance .
2.2 Effect virgin coconut oils on methane gas concentration
In another study, to streamline the digestive process in the rumen, Suryani
The purpose of this experiment is to get the best VCO level combined with the best type of DFM stage 1 on ammoniated palm fronds. The three VCO levels tested were 2, 3, and 4% DM. The two best types of DFM from stage 1 used as controls were
|Treatments||CH4 (ml/g DM)||Protozoa (cell/mL-1)|
|P1: SC 1% + 0% VCO||21.74 ± 1.16||7.08 x 104 ± 0.23|
|P2: SC 0.5% + BA 0.5% + 0% VCO||22.94 ± 0.84||7.23 x 104 ± 0.36|
|P3: SC 1% + 2% VCO||11.78 ± 0.62||1.92 x 104 ± 0.09|
|P4: SC 0.5% + BA 0.5% + 2% VCO||12.92 ± 0.22||2.23 x 104 ± 0.09|
|P5: SC 1% + 3% VCO||11.87 ± 0.79||1.97 x 104 ± 0.09|
|P6: SC 0.5% + BA 0.5% + 3% VCO||12.58 ± 0.15||2.65 x 104 ± 0.15|
|P7: SC 1% + 4% VCO||12.75 ± 0.93||3.38 x 104 ± 0.09|
|P8: SC 0.5% + BA 0.5% + 4% VCO||13.49 ± 0.09||3.28 x 104 ± 0.15|
The results of the orthogonal polynomial test show a quadratic relationship (P < 0.05) between the level of VCO (X, %) and the concentration of methane gas in the rumen (Y, mM) with the equation y = 1.2682x2–7.3169 + 22.281 and the coefficient of determination R2 = 0.98137 (Figure 3).
Based on the orthogonal polynomial test, methane gas concentration at the level of 2% VCO addition with DFM
The results of the orthogonal polynomial test give a quadratic relationship between the VCO level (X, %) and the protozoa population (Y, cell/mL-1), the Eq. Y = 0.7546x2–3.9464x + 7.1323 and the coefficient of determination (R2) = 0.98564 is shown on Figure 4. The average population of protozoa with the addition of VCO in the rumen can be seen in Table 3.
|DM Intake (kg day−1)||3.16a||3.01b||2.99b||2.57c||0.143|
|DM/BW0.75 (g kg −1 b.wt.0.75 d−1)||79.94a||75.68ab||74.24ab||67.36b||1.790|
|OM Intake (kg/h/d)||3.93a||3.74b||3.72b||3.19c||0.017|
|OM /BW0.75 (g kg −1 b.wt.0.75 d−1)||99.28a||97.14a||93.98a||83.65b||1.504|
|ADG (kg day−1)||0.53c||0.63b||0.63b||0.71a||0.007|
|Feed Efficiency (%)||16.96c||20.84b||21.34b||27.77a||0.311|
|Methane production (L day−1)||109.01c||103.27b||102.61b||86.52a||0.501|
|Nitrogen intake (g day−1)||59.20a||56.34b||55.96b||47.84c||0.815|
|Nitrogen retention (g day−1)||50.51a||47.99a||47.16a||37.23b||0.797|
Based on the orthogonal polynomial test, the protozoa population decreased with VCO supplementation. Supplementation of 2% and 3% VCO (P3,P4,P5,P6) on palm fronds with the addition of DFM
Meanwhile, total protozoa (especially
In other studies, Suryani
DFM and VCO supplementation decreased methane production by 5.26, 5.87, and 20.63% respectively. The highest ration efficiency was in DFM
Therefore, to reduce hydrogen production to methane, hydrogen must be switched to propionate production via lactate or fumarate . H2 and CO2 are substrates used to form methane. According to Wilkie  the role of hydrogen in the methane production process is as a source of electrons, so the low level of H2 in the rumen is an indication of activity using H2 to reduce CO2 to CH. In addition, to form one mole of CH4 requires four moles of H2. The rate of H2 utilization is four times the rate of methane production so that H2 in the rumen never accumulates. The following is the stoichiometry of the carbohydrate fermentation reaction in producing methane gas in the rumen:
The effect of DFM and VCO supplementation on Bali cattle on blood profile can be seen in Table 4.
The results showed that DFM and VCO supplementation had a very significant effect (p < 0.05) in reducing cholesterol, LDL and increasing HDL blood levels of Bali cattle. DFM and VCO supplementation had no significant effect (P > 0.05) on triglycerides, urea, protein, albumin, and glucose. VCO contains MCFA, which is a saturated fatty acid (Figure 1), its addition in the ration if consumed by livestock can help lower cholesterol because of the nature of this fatty acid, which can be absorbed directly by the animal’s body so that it does not cause fat accumulation that causes cholesterol. This is supported by Fernando
This study can conclude that individual
3. Effect of different source tannins on methane gas production
Bioactive compounds, including polyphenols, carotenoids, omega-3 fatty acids, vitamins, organic acids, nucleotides, and nucleosides, have attracted significant attention for their role in preventing several chronic diseases in humans. In animal husbandry, especially ruminant nutrition, bioactive plant polyphenolic compounds such as tannins and saponins have been studied extensively for optimizing bioprocesses in the rumen through feed manipulation. Manipulation of feed using tannins as an agent of rumen defaunation is one way to overcome global climate change due to the effects of greenhouse gases, one of which is caused by methane gas from ruminants . Feeds containing tannins will be anti-nutrients that limit livestock production when the crude protein concentration in the feed is high because it can reduce the absorption of amino acids . Tannins can also cause poisoning if consumed by livestock in excess, and there are many
|Treatments||DM (%)||OM (%)||Protozoa population (cell mL-1)||CH4 (ml/g DM)||VFA Total (mM)||A: P Ratio|
|T0||48.45b||51.34b||11.43 x 104a||27.22a||71.00b||3.98a|
|B1||51.59ab||54.17ab||2.3 x 104c||23.64ab||83.70ab||2.70c|
|B2||52.09a||57.30a||1.4 x 104c||12.67c||95.78a||3.52ab|
|B3||50.93ab||53.15ab||4.8 x 104b||13.14c||65.94b||3.38ab|
|C1||51.08ab||54.16ab||4.7 x 104a||15.13c||75.49ab||2.58b|
|C2||50.69ab||52.83ab||9.3 x 104b||17.12c||79.40ab||3.65a|
|C3||48.65b||51.04b||8.8 x 104a||21.90b||62.44b||3.40b|
|A||58,83c||59.50c||6.3 x 105a||22.72a||72.00||2.14b|
|B||62.5b||63.72b||5.8 x 105b||21.46b||74.25||1.50a|
|C||66.33a||68.66a||4.9 x 105c||16.27c||75.45||1.70a|
|D||68.54a||69.50a||4.7 x 105c||14.14c||76.8||1.33a|
The results showed that different sources and doses of tannins proved to have different effects on decreasing methane production . The
The addition of
The potential of plant bioactive compounds such as tannins and saponins as defaunation agents and reducing methane emissions can be combined with direct-fed microbes. There is not much literature on decreasing methane production that combines the two in
In conclusion, the overall reduction in methane production in agriculture by-products as feed-based beef cattle can be made by improving feed quality through a combination of processing techniques and efforts to optimize bioprocesses in the rumen, which include supplementation of feed additives such as direct-fed microbials, methanogenesis inhibitors and plant bioactive compounds. Supplementation of DFM type
Thank to the Ministry of Research and Technology, and Andalas University that providing grants to support the research.