The Effect of Temulawak ( Curcuma xhantorrhiza ) Dose on Tofu Dreg Plus Complete Feed Silage With Methane Relative Mitigation: In-Vitro

. Temulawak is an antimicrobial herb that can preserve the proteins in the feed from deamination and decay during the silage period. Tofu dregs are a feed ingredient that is difficult to preserve because they have high water content and protein. Temulawak powder may decrease deamination during the silage process because it has the ability as an antimicrobe and protein binder. Temulawak can reduce methane emissions by diminishing the number of protozoa that serve as hosts for methanogenic archaea. Temulawak with a harvest age of 3 months after planting, then dried at 65°C to constant weight, grounded, and tested on mix ration silage at doses of 0%, 0.25%, 0.5%, 0.75 %, and 1% from feed dry matter (DM). The experiment was carried out four times using a complete random design and in vitro silage was evaluated in the following 14 days. The results indicated that adding temulawak would reduce ammonia production with p<0.01, significantly increase the crude protein content of silage P<0.05 and decrease rumen ammonia P<0.01. The treatment increased total volatile


Introduction
The process of protein deamination occurs when a wet ingredient with high protein is stored with the silage method.De-amination is accomplished by spoilage bacteria such as Clostridium sp, E. coli, and others [1].Curcumin, which is present in Temulawak powder, has antimicrobial properties [2].Tannin in temulawak also can bind protein and decrease the deamination proses [3].Consequently, temulawak powder can act as a protein-protecting agent during the tofu dreg silage process.
Microbes in the rumen ferment feed on an anaerobic system, on the other hand, the process produces methane [4].The consumption of crude fiber in ruminants is closely related to the production of methane [5].The feeding quality of ruminants affects their methane production [6].Reducing the enteric fermentation in the rumen has environmental benefit and increase livestock production [7] Manipulation of rumen metabolism by diminishing the number of microbes, particularly methanogenic archaea, using bioactive compounds is one method to decrease methane enteric rumen fermentation [8].Methane can be reduced by decreasing the protozoa population as hosts for methanogenic archaea [9].One of the ingredients that can be added to feed is temulawak.Because these plants contain bioactive compounds with antimicrobial properties [10] [11].
Temulawak addition not only reduces methane production but also increases livestock production and health.Because temulawak contains phenols, curcumin, essential oils, and antioxidants [12], it has anti-inflammatory, antimicrobial, anti-cancer, and anti-tumor properties [13].Temulawak may replace AGP (antibiotic as a growth promoter) due to its capacity as an immunity booster [14].

Preparation Temulawak Powder
Fresh Temulawak from Sinar Sari village, Dramaga District, Bogor was harvested after 3 months of planting, The temulawak was chopped to 3 mm thickness and dried in an oven with a temperature of 65 0 C until reached a stable weight.The dried temulawak grinder by 40 mesh filter.

Silage prosses
Study about temulawak powder dosage on complete feed plus tofu dreg silage using fresh comparison 1:1.Tofu dregs were obtained freshly from home industrial tofu at Dramaga District, Bogor.The complete feed using commercial products from Agro Apis Palacio Company at Bubulak.The formulation of feed ingredients in the table 1:

In-Vitro analysis procedure
The In-vitro analysis of complete feed plus tofu dreg silage was preserved for 14 days.The silage mixture dried at oven 65 0 C for 24 hours.The Invitro study using the Theodorou model 1994 [15].In-vitro fermentation using rumen fluid inoculum was collected at a slaughterhouse in Bubulak, Bogor City (part of the Bogor City Agriculture Service) so animal welfare is noticed in this area.The dried sample approximately 500 mg was inserted into a 125 ml bottle and added with rumen fluid and buffer, each of them 15 and 60 ml, respectively.Treatments in the incubation bottles were according to a randomized complete design.Incubation was carried out in four replicates (four bottles per replicate).The bottles were sealed with butyl rubber stoppers and aluminum crimps.Incubation was carried out for 24 h at 39ºC.Gas production was recorded and collected in the serum bottles during the incubation.The gas was tested amount of methane with a methane meter at the Indonesian Soil Research Institute.After 24 h, the supernatant was collected for determination of total volatile fatty acids (VFA) and ammonia concentration [16].The residue was weighed to determine in vitro dry matter digestibility and in vitro organic matter digestibility.

Data Analysis
This study uses a randomized complete design that is obtained as follows: P0: Complete Feed+ Tofu dregs P1: P0 + temulawak 0,25% from DM P2: P0 + temulawak 0.50% from DM P3: P0 + temulawak 0.75% from DM P4: P0 + temulawak 1.0% from DM Every treatment was repeated in 4 replications.So, the mathematical model was obtained: Yij = μ + αi + €ij Information: Yij = Parameter values affected by temulawak dosage μ = General Mean αi = The effect of temulawak dosage on parameter values €i = The Error of study The data has been analyzed with analysis of variant ANOVA using SPSS 16.0.If any significant differences between factor have been tasted with Duncan's analysis.

In vitro Effect of Temulawak Powder Dosage on Rumen Metabolism
During the rainy season, one of the most common challenges is the preservation of high-protein and feed ingredients [17].In the silage process, spoilage bacteria such as Clostridium sp, Escherichia coli, and others perform the de-amination process [1].De-amination process transforms the amin chain in the protein into ammonia, which increases the difficulty of lowering the silage pH [1].Temulawak is a rhizome that has antibacterial properties [13].Temulawak powder was able to prevent protein degradation during the silage-making procedure.
There are deaminating microorganisms in the rumen that can convert feed proteins into ammonia and carbon groups [3].The rumen can convert non-protein nitrogen compounds into microbial proteins that can be consumed by the host [18].Therefore, feed protein must be protected from microorganisms capable of deaminating protein.In addition to the deamination process in the anaerobic rumen, methane will be emitted into the atmosphere, which is one of the causes of climate change [4].Methane is a greenhouse gas that can absorb 20 times more heat than carbon dioxide [19].Table 2 displays the test parameters for rumen metabolites.Protein is an essential nutrient for the development of livestock's average daily gain.The amount of protein in fodder has a significant effect on the weight gain of livestock.The presence of elevated levels of ammonia in the incubation fluid in vitro reveals protein damage in the rumen [3].Ammonia in the rumen results from the deamination of proteins by rumen microorganisms.The addition of temulawak significantly decreased the level of ammonia in the rumen P<0.01.Reduced deaminator microorganisms in the presence of active compounds in temulawak can facilitate the reduction of ammonia [20].Tannins in temulawak can operate as a protein bypass agent, leading to a reduction in ammonia levels [3].
The pH value of the rumen is strongly correlated with the health of its digestive organs and the equilibrium of its feed formulations.The addition of temulawak powder revealed a normal range in the complete diet plus silage tofu dregs test.The addition of 1% DM of Curcuma powder substantially decreased the rumen pH, which may have been caused by the ammonia produced by the breakdown of feed protein because of decreased deamination.Due to the active compounds of temulawak reducing the number of pathogenic bacteria such as Coxidia Sp [21].The bond between protein and tannin in temulawak can also reduce ammonia [1].Therefore, it can be concluded that temulawak is capable of protecting proteins during the silage process and in the rumen.
The primary source of energy for ruminants is volatile fatty (VFA), which are converted to glucose in the liver [22].A high VFA value indicates that the process of converting feed into energy in the rumen is functioning correctly.The dose-dependent effect of temulawak powder on VFA levels was statistically significant (P<0.01).The optimal addition of VFA occurred when the temulawak powder dose was 0.25 percent of DM.Due to a decrease in protozoa that consume fiber-digesting bacteria, the subsequent increase in VFA values [23].In general, the addition of 0.25 percent dry matter (DM) of temulawak powder to complete feed plus tofu dreg silage remnants is the optimal dosage for VFA production.
In addition to in vitro estimation of VFA values, the digestibility of the feed is an indicator of the efficacy of a feed treatment or the optimal feed formulation for animals.On the rumen and post-rumen organs, in vitro, digestibility measurements can be conducted.The digestibility of feed formulations dry matter, organic matter, and gas total production was determined.The dosage of temulawak powder has no effect on dry matter, organic matter digestibility, and total gas production P>0.05.
Methane is a gas that can absorb 20 heat higher than carbon dioxide [24].Methane is produced during the electron transfer process when the mitochondrial environment is in an anaerobic state, so methane formation will always occur in anaerobic rumens [7].The addition of 0.25 percent dry matter (DM) of temulawak powder to feed significantly decreased methane P<0.01.[8] Plants contain bioactive compounds that can reduce the quantity of protozoa.Since the majority of archaea methanogen live in symbiosis with protozoa [25] the decline in protozoa is closely related to mitigating methane gas.
This study compared the methane content with several parameters that affect livestock growth, including the amount of feed dry matter, the amount of feed organic matter, total VFA, and total gas production.This comparison demonstrates that the addition of temulawak powder at a concentration of 0.25 percent had the greatest effect on methane.The concentration of temulawak powder significantly decreased methane relative to organic matter, total VFA, and total gas with a P< 0.01.The inclusion of temulawak powder revealed that the addition of 0.25 and 0.5 percent doses of dry matter was less effective than other treatments with a P<0.05.The ability of bioactive compounds in temulawak to diminish protozoa is correlated with the decrease in methane [25].Protozoa not only hosts methanogen archaea but also consume bacteria that ferment the feed.Due to decreasing protozoa, it can increase the digestibility of dry matter, organic matter, and VFA which are then converted into energy in the livers of ruminants.

Conclusion
The addition of 1% dry matter temulawak powder was effective in reducing the risk of protein damage during the optimal silage preservation method.The addition of 0.25% temulawak powder to complete feed plus tofu dreg silage had a positive effect on total VFA production while protecting feed protein from deamination.This level is optimal for reducing absolute methane, methane relative to dried matter, organic matter, VFA total, and total gas.

Table 2 .
Feed Nutrient Analysis

Table 3 .
Effect of temulawak powder concentrations on rumen metabolism (in-vitro) after silage process

Table 4 .
Effect of temulawak powder concentration on methane relative