This study focuses on the feasibility of bio-gas production using anaerobic digestion by measuring methane generation and biodegradability through the BMP test of industrial organic wastes. Organic wastes consist of entrails of pigs and organic residues of rumen generated from slaughter houses, wastewater sludge from slaughter waste water, fish offal and residues of vegetables from public wholesale markets, and wastewater sludge from the process of wastewater treatment in paper mill. The cumulative methane production by BMP test ranges from 149.3 ml/g-VS to 406.6 ml/g-VS and this is similar to methane generation of the normal wastewater sludge and food waste. As a result of measurement of biodegradability, wastewater sludge (S1 ~ S4) is low, ranging from 27.1% to 58.9 % and organic residues of rumen (G1) is low at 49.6 %. In conclusion, it turned out that raising the hydrolysis by various pre-treatments is necessary in order to produce bio-gas by using industrial organic wastes.
The objective of this study was to determine the effects of protein sources and roughage (R) to concentrate (C) ratio on in vitro fermentation parameters using a gas production technique. The experimental design was a $2{\times}5$ factorial arrangement in a completely randomized design (CRD). Factor A was 2 levels of protein sources yeast fermented cassava chip protein (YEFECAP) and soybean meal (SBM) and factor B was 5 levels of roughage to concentrate (R:C) ratio at 80:20, 60:40, 40:60, 20:80, and 0:100, respectively. Rice straw was used as a roughage source. It was found that gas production from the insoluble fraction (b) of YEFECAP supplemented group was significantly higher (p<0.05) than those in SBM supplemented group. Moreover, the intercept value (a), gas production from the insoluble fraction (b), gas production rate constants for the insoluble fraction (c), potential extent of gas production (a+b) and cumulative gas production at 96 h were influenced (p<0.01) by R:C ratio. In addition, protein source had no effect (p>0.05) on ether in vitro digestibility of dry matter (IVDMD) and organic (IVOMD) while R:C ratio affected the IVDMD and IVOMD (p<0.01). Moreover, YEFECAP supplanted group showed a significantly increased (p<0.05) total VFA and $C_3$ while $C_2$, $C_2:C_3$ and $CH_4$ production were decreased when compared with SBM supplemented group. In addition, a decreasing R:C ratio had a significant effect (p<0.05) on increasing total VFA, $C_3$ and $NH_3$-N, but decreasing the $C_2$, $C_2:C_3$ and CH4 production (p<0.01). Furthermore, total bacteria, Fibrobacter succinogenes, Ruminococcus flavefaciens and Ruminococcus albus populations in YEFECAP supplemented group were significantly higher (p<0.05) than those in the SBM supplemented group while fungal zoospores, methanogens and protozoal population remained unchanged (p>0.05) as compared between the two sources of protein. Moreover, fungal zoospores and total bacteria population were significantly increased (p<0.01) while, F. succinogenes, R. flavefaciens, R. albus, methanogens and protozoal population were decreased (p<0.01) with decreasing R:C ratio. In conclusion, YEFECAP has a potential for use as a protein source for improving rumen fermentation efficiency in ruminants.
Solubilization pretreatments were conducted to enhance the anaerobic digestion of the waste activated sludge. Four pretreatment techniques including heating, sonication freezing and thawing, and enzyme addition were employed to solubilize the waste activated sludge under various conditions. Thermal pretreatment by heating showed the highest efficiency compared with other methods, and freezing and thawing was confirmed as a feasible alternative of solubilization as well as the pretreatment of dewatering. There is a clear correlation between the solubilization efficiency of the waste activated sludge and the gas production. Batch digestion results showed the cumulative gas production as much as four times after thermal pretreatment as compared with that by the control sludge without pretreatment. As a result, hydrolysis or solubilization pretreatment might play a significant role in the high rate digestion of the waste activated sludge.
The suppressive effect of monensin as an ionophore-feed additive on enteric methane (CH4) emission and renewable methanogenesis were evaluated. To clarify the suppressive effect of monensin a respiratory trial with head cage was performed using Holstein-Friesian steers. Steers were offered high concentrate diets (80% concentrate and 20% hay) ad libitum with or without monensin, galacto-oligosaccharides (GOS) or L-cysteine. Steers that received monensin containing diet had significantly (p < 0.01) lower enteric CH4 emissions as well as those that received GOS containing diet (p < 0.05) compared to steers fed control diets. Thermophilic digesters at 55℃ that received manure from steers fed on monensin diets had a delay in the initial CH4 production. Monensin is a strong inhibitor of enteric methanogenesis, but has a negative impact on biogas energy production at short retention times. Effects of the activity of coprophagous insects on CH4 and nitrous oxide (N2O) emissions from cattle dung pats were assessed in anaerobic in vitro continuous gas quantification system modified to aerobic quantification device. The CH4 emission from dungs with adults of Caccobius jessoensis Harold (dung beetle) and the larvae of the fly Neomyia cornicina (Fabricius) were compared with that from control dung without insect. The cumulative CH4 emission rate from dung with dung insects decreased at 42.2% in dung beetles and 77.8% in fly larvae compared to that from control dung without insects. However, the cumulative N2O emission rate increased 23.4% in dung beetles even though it reduced 88.6% in fly larvae compared to dung without coprophagous insects. It was suggested that the antibacterial efficacy of ionophores supplemented as a growth promoter still continued even in the digested slurry, consequently, possible environmental contamination with the antibiotics might be active to put the negative impact to land ecosystem involved in greenhouse gas mitigation when the digested slurry was applied to the fields as liquid manure.
Objective: The present study was conducted to examine the gas production, fermentation characteristics, nutrient degradation, and methanogenic community composition of a rumen fluid culture with Broussonetia papyrifera (B. papyrifera) subjected to ensiling or steam explosion (SE) pretreatment. Methods: Fresh B. papyrifera was collected and pretreated by ensiling or SE, which was then fermented with ruminal fluids as ensiled B. papyrifera group, steam-exploded B. papyrifera group, and untreated B. papyrifera group. The gas and methane production, fermentation characteristics, nutrient degradation, and methanogenic community were determined during the fermentation. Results: Cumulative methane production was significantly improved with SE pretreatment compared with ensiled or untreated biomass accompanied with more volatile fatty acids production. After 72 h incubation, SE and ensiling pretreatments decreased the acid detergent fiber contents by 39.4% and 22.9%, and neutral detergent fiber contents by 10.6% and 47.2%, respectively. Changes of methanogenic diversity and abundance of methanogenic archaea corresponded to the variations in fermentation pattern and methane production. Conclusion: Compared with ensiling pretreatment, SE can be a promising technique for the efficient utilization of B. papyrifera, which would contribute to sustainable livestock production systems.
The objective of this study was to determine the roughage to concentrate (R:C) ratio with rain tree pod meal (RPM) supplementation on in vitro fermentation using gas production technique. The experiment design was a 6${\times}$4 factorial arrangement in a CRD. Factor A was 6 levels of R:C ratio (100:0, 80:20, 60:40, 40:60, 20:80 and 0:100) and factor B was 4 levels of RPM (0, 4, 8 and 12 mg). It was found that gas kinetic, extent rate (c) was linearly increased (p<0.01) with an increasing level of concentrate while cumulative gas production (96 h) was higher in R:C of 40:60. In addition, interaction of R:C ratio and RPM level affected $NH_3-N$ and IVDMD and were highest in R:C of 0:100 with 0, 4 mg of RPM and 40:60 with 8 mg of RPM, respectively. Moreover, interaction of R:C ratio and RPM level significantly increased total volatile fatty acids and propionate concentration whereas lower acetate, acetate to propionate ratios and $CH_4$ production in R:C of 20:80 with 8 mg of RPM. Moreover, the two factors, R:C ratio and RPM level influenced the protozoal population and the percentage of methanogens in the total bacteria population. In addition, the use of real-time PCR found that a high level of concentrate in the diet remarkably decreased three cellulolytic bacteria numbers (F. succinogenes, R. flavefaciens and R. albus). Based on this study, it is suggested that the ratio of R:C at 40:60 and RPM level at 12 mg could improve ruminal fluid fermentation in terms of reducing fermentation losses, thus improving VFA profiles and ruminal ecology.
Dietary fatty acid including mainly palmitic acid and stearic acid was fed to fattening cattle and its effect on body weight gain, plasma lipid contents and rumen liquid fermentation in vitro was examined. In expt. 1, the effect of dietary fatty acid on body weight gain and plasma lipid concentrations was examined. In the control diet group, cattle were fed 1 kg/day of rice straw and concentrate which satisfied the requirement. In the fatty acid group, cattle were given 250 g/d of fatty acid with the same diet of the control diet group. In the excess concentrate group, cattle were given the same diet of the control diet group plus 735 g/d of concentrate corresponding to the same TDN of 250 g/d of fatty acid. Diets were given for 7 days. Body weight gain of cattle given dietary fatty acid was significantly greater than that of cattle fed only rice straw and concentrate. When dietary fatty acid was added to cattle feed, plasma NEFA and HDL-cholesterol concentrations increased. In expt. 2, the influence of dietary fatty acid on gas production and VFA profile in the rumen liquid was investigated in vitro. In the control group, 10 mg of rice straw and 90 mg of concentrate were incubated in the rumen fluid. In the excess concentrate group, 10 mg of rice straw and 97.5 mg of concentrate were incubated. In the fatty acid group, 10 mg of rice straw, 90 mg of concentrate and 2.5 mg of fatty acids were incubated. The rumen liquid mixed with feed materials was incubated for 24 h and the cumulative gas volume was measured. The VFA profile was also measured. Cumulative gas volume in the rumen liquid with fatty acid was equal to the control. Excess concentrate increased cumulative gas volume compared to the fatty acid group. There was no significant difference in total VFA concentration between experimental diet groups. It is suggested that dietary fatty acid has the potency to improve growth performance in fattening cattle without failure in rumen fermentation.
In order to develop a high cellulolytic direct-fed microorganism (DFM) for ruminant productivity improvement, this study isolated cellulolytic bacteria from the rumen of Holstein dairy cows, and compared their cellulolytic abilities via DM degradability, gas production and cellulolytic enzyme activities. Twenty six bacteria were isolated from colonies grown in Dehority's artificial (DA) medium with 2% agar and cultured in DA medium containing filter paper at $39^{\circ}C$ for 24h. 16s rDNA gene sequencing of four strains from isolated bacteria showed that H8, H20 and H25 strains identified as Ruminococcus flavefaciens, and H23 strain identified as Fibrobacter succinogenes. H20 strain had higher degradability of filter paper compared with others during the incubation. H8 (R. flavefaciens), H20 (R. flavefaciens), H23 (F. succinogenes), H25 (R. flavefaciens) and RF (R. flavefaciens sijpesteijn, ATCC 19208) were cultured in DA medium with filter paper as a single carbon source for 0, 1, 2, 3, 4 and 6 days without shaking at $39^{\circ}C$, respectively. Dry matter degradability rates of H20, H23 and H25 were relatively higher than those of H8 and RF since 2 d incubation. The cumulative gas production of isolated cellulolytic bacteria increased with incubation time. At every incubation time, the gas production was highest in H20 strain. The activities of carboxymethylcellulase (CMCase) and Avicelase in the culture supernatant were significantly higher in H20 strain compared with others at every incubation time (p<0.05). Therefore, although further researches are required, the present results suggest that H20 strain could be a candidate of DFM in animal feed due to high cellulolytic ability.
Marine biomass is considered an important substrate for anaerobic digestion to recovery energy i.e. methane. Nevertheless, marine biomass has attracted little attention by researchers compared to terrestrial feedstock for anaerobic digestion. In this study, biochemical methane potential (BMP) test was used to evaluate generation of renewable energy from starfish. A cumulative biogas yield of $748{\pm}67mL\;g^{-1}VS^{-1}$ was obtained after 60 days of digestion. The cumulative methane yield of $486{\pm}28mL\;CH_4\;g^{-1}VS^{-1}$ was obtained after 60 days of digestion. The methane content of the biogas was approximately 70%. The calculated data applying the modified Gompertz equation for the cumulative $CH_4$ production showed good correlation with the experimental result obtained from this batch study. Since the result obtained from this study is comparable to results with other substrates, marine biomass can be co-digested with food waste or swine wastewater to produce $CH_4$ gas that will help to reduce the gap in global energy demand.
The present study showed the advantages of dried Bacillus subtilis culture (DBSC) supplementation on reducing ammonia gas release in the poultry house. In Experiment 1, 65-week-old Hyline W-36 hens were raised in individual wire-floor cages in a windowless house, and divided into two groups of 180 hens each. One group was fed diets without DBSC as the control and another group was fed a diet supplemented with 2% DBSC. In Experiment 2, 2-week-old broiler chicks were divided into 3 treatment groups of 20 chicks each and maintained in individual floor cages. One group was fed the diet without DBSC and other two groups were fed the diet supplemented with 1 or 2% DBSC, respectively. In experiment 1, DBSC consistently reduced ammonia gas release in the laying house (p<0.01) and manure storage facilities (p<0.01). incubation of feces for 1, 2, 3, 4, 5, 6, 24 or 48 hours showed that DBSC consistently reduced ammonia gas release. In Experiment 2, DBSC reduced ammonia gas release in the broiler house; however, DBSC had no effect on total N, urate-N and ammonia-N contents of feces, but it improved cumulative N utilization and decreased serum urea-N concentration when chicks when chicks were fed 1% DBSC.
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