Maize-soybean meal diets with 0, 100, 200 and 300 g/kg double zero rapeseed meal ('00' RSM) with and without an enzyme mixture (xylanase, pectinase, cellulase) at a level of 1.6 g/kg were evaluated with 624 day-old broiler chicks for 5 weeks. The birds were randomly allocated to eight dietary treatments with three replicates of 26 birds each. Average daily gain (ADG) and feed intake (FI) were recorded weekly and ileal viscosity, organ weights, serum enzyme activity, hormonal profile and hematological parameters were measured at the end of week 5. Average daily gain during the weekly periods was significantly influenced by the dietary level of '00'RSM (p<0.01). Inclusion of '00' RSM improved the ADG up to day 28 with the increased level; beyond that time no improvement was recorded when compared to control groups. However, ADG from 1-35 days was significantly different between 300 g/kg inclusion level of '00' RSM and the control diet. Inconsistent decline in feed intake and feed conversion ratio was observed up to day 21 and the trend was reversed thereafter. The proportion of '00' RSM in the diet had a significant ($p{\leq}0.05$) influence on thyroid weight but had no effect on the relative weights of liver and heart, serum enzyme activities (${\gamma}$-glutamyl transferase, alanine amino transferase and aspartate amino transferase), thyroid hormones ($T_3$ and $T_4$), hemoglobin level and hematocrit. Significant improvement in ADG was recorded during the 2nd week of age with the addition of enzyme, whereas for all other periods, including the whole period of the trial, higher but non-significant ADG was observed. FI and FCR were not affected by the addition of enzyme but there was a numerical reduction in FCR during the whole period. The addition of enzyme reduced the ileal viscosity at all levels of '00' RSM inclusion. The results suggest that '00' RSM can be included up to 300 g/kg in broiler diets without any adverse effects on health and performance. The addition of commercial enzyme mixture containing xylanase, pectinase, cellulase to broiler diets containing '00'RSM has some effect on growth rate and feed conversion efficiency.
Kim, Chang-Hyun;Park, Joong Kook;Lee, Gi Yeong;Seo, In Joon
Asian-Australasian Journal of Animal Sciences
/
v.18
no.10
/
pp.1435-1439
/
2005
Bacterio-mineral water (BMW) produced from manure has been known to exert a number of positive effects on animal production and odor control. An experiment was conducted to examine the effects of BMW produced from bio-reacted swine manure on in vitro gas production, cellulose degradation, microbial growth and fibrolytic enzyme activities of mixed rumen microorganisms. The five levels of 0, 0.001, 0.005, 0.01 and 1.0% BMW were supplemented into serum vials containing mixed rumen microorganisms. Incubations were carried out anaerobically at $39^{\circ}C$ without shaking for 0, 12, 24, 48, 72 and 96 h. There were no significant (p>0.05) differences among the treatments for the initial rate of gas production. At 72 h incubation, the gas production tended (p<0.1) to be increased by the 0.01 and 1.0% BMW treatments compared with control and the 0.001% BMW treatment. At the end of incubation (96 h), the sample supplemented with 0.01% BMW was higher (p<0.05) than control (0% BMW) in the gas production. The microbial growth rate was increased by all the BMW treatments, while 0.01% BMW was most effective in stimulating the growth rate. Although the addition of BMW on the filter paper DM degradation was not significantly influenced throughout the incubation period except the 48 h incubation, DM degradation tended to be increased by all BMW treatments compared with control. The addition of both 0.005 and 0.01% BMW highly increased (p<0.05) CMCase activity compared with control after 24 h and 48 h incubation, while at the 72 h incubation the 0.01% BMW addition only significantly increased (p<0.05). After 72 h incubation, the xylanase activity was significantly (p<0.05) increased with the addition of 1.0% BMW compared with the addition of 0.001 and 0.005% BMW, while at the other incubation times, the xylanase activity was not different among the treatments. In conclusion, the 0.01% BMW of supplementation level would be the suitable addition level to stimulate rumen fermentation increasing microbial growth and cellulose degradation.
Yam has been recognized as healthy food due to its various biological activities, such as anti-obesity, antimicrobial, anticancer and immuno-stimulation activities, and its consumption has been increased during last decades. In this study, to investigate low-temperature, long-term storage of yam and to develop processed yam products, yam-putrefactive psychrotrophic bacteria were isolated from rotted yam and identified based on BBL identification system, fatty acid analysis in cell membrane and 16S rDNA sequence analysis. The putrefaction activity of isolated thirteen bacteria was evaluated using yam-slices (NaOCl-treated, autoclaved yam and without treatment), and YAM-10 and YAM-12 were identified as major psychrotrophic putrefactive bacteria. Both YAM-10 (Pseudomonas cepacia) and YAM-12 (Pseudomonas rhodesiae) bacteria grew well at 4$\sim$12$^{\circ}C$ and showed strong activity of polymer degrading enzymes, especially amylase, carboxy methyl cellulase and xylanase, at 20$^{\circ}C$. But they failed to grow at acidic pH (<5) or alkaline pH (>10). Our results suggested that the control of psychrotrophic Pseudomonas sp. by pH change and inhibition of polymer degrading enzymes, such as amy-lase, are necessary to long-term storage of yam.
In order to investigate the compositional change of the lauan and pine sawdust media during the coltivation of oyster mushroom (Pleurotus ostreatus) chemical components of the two media and mushroom were analized periodically from spawning to ending. The results are summerized as follows: 1) The total yield of mushroom from the lauan sawdust media was 6.39 kg/$m^2$, and from the Pine saw-dust media 6.31 kg/$m^2$, 94% of which was produced from the first and second cropping period. 2) In both media ash content increased gradually, while organic matter content decreased as the cultivation progressed. Moisture content, which was about 79% at the time of spawning, decreased a little during the priod of mycelial propagation, after which no change was observed. 3) Total nitrogen content of the two media decreased gradually as the cultivation progressed, and total loss of insoluble nitrogen was greater than that of soluble nitrogen. Amino nitrogen content tended to increase continually. 4) In both media pentosan, $\alpha$-cellulose, lignin and C/N ratio decreased gradually, while the content of reduced sugar, trehalose and mannitol continued to increase. 5) In both media phosphorus, potassium, manganese and zinc decreased, and magnecium, calcium, iron and copper showed irregular changes. 6) CMC-saccharifying and liquefying activity gradually increased from after mycelial propagation to the second cropping, after which it decreased in both media. Xylanase activity rapidly and greatly increased during the second cropping period rather than the first period. At the start of the third cropping period the activity decreased slowly. Protease activity was highest after mycelial propagati, after which it gradually decreased. The pH of the two media decreased as the cultivation progressed. 7) In both media little change was observed in the content of the components of mushroom cropped from the first to the third period, but slight decrease was noticed at the fourth cropping.
Biomass is originally photosynthesized from inorgainic compounds such as $CO_2$, minerals, water and solar energy. Recent studies have shown that anaerobic bacteria have the ability to convert recalcitrant biomass such as cellullosic or chitinoic materials to useful compounds. The biomass containing agricultural waste, unutilized wood and other garbage is expected to utilize as feed, food and fuel by microbial degradation and other metabolic functions. In this study we isolated several anaerobic, cellulolytic and chitinolytic bacteria from rumen fluid, compost and soil to study their related enzymes and genes. The anaerobic and cellulolytic bacteria, Clostridium thermocellum, Clostridium stercorarium, and Clostridium josui, were isolated from compost and the chitinolytic Clostridium paraputrificum from beach soil and Ruminococcus albus was isolated from cow rumen. After isolation, novel cellulase and xylanase genes from these anaerobes were cloned and expressed in Escherichia coli. The properties of the cloned enzymes showed that some of them were the components of the enzyme (cellulase) complex, i.e., cellulosome, which is known to form complexes by binding cohesin domains on the cellulase integrating protein (Cip: or core protein) and dockerin domains on the enzymes. Several dockerin and cohesin polypeptides were independently produced by E. coli and their binding properties were specified with BIAcore by measuring surface plasmon resonance. Three pairs of cohesin-dockerin with differing binding specificities were selected. Two of their genes encoding their respective cohesin polypeptides were combined to one gene and expressed in E. coli as a chimeric core protein, on which two dockerin-dehydrogenase chimeras, the dockerin-formaldehyde dehydrogenase and the dockerin-NADH dehydrogenase are planning to bind for catalyzing $CO_2$ reduction to formic acid by feeding NADH. This reaction may represent a novel strategy for the reduction of the green house gases. Enzymes from the anaerobes were also expressed in tobacco and rice plants. The activity of a xylanase from C. stercorarium was detected in leaves, stems, and rice grain under the control of CaMV35S promoter. The digestibility of transgenic rice leaves in goat rumen was slightly accelerated. C. paraputrificum was found to solubilize shrimp shells and chitin to generate hydrogen gas. Hydrogen productivity (1.7 mol $H_2/mol$ glucos) of the organism was improved up to 1.8 times by additional expression of the own hydrogenase gene in C. paraputrficum using a modified vector of Clostridiu, perfringens. The hydrygen producing microflora from soil, garbage and dried pelletted garbage, known as refuse derived fuel(RDF), were also found to be effective in converting biomass waste to hydrogen gas.
Kim, Hyun-Joo;Cho, Eun-Jin;Lee, Kwang-Ho;Kim, Su-Bae;Bae, Hyeun-Jong
Journal of the Korean Wood Science and Technology
/
v.39
no.1
/
pp.95-103
/
2011
We have investigated pretreatment of waste wood using milling refinery combined with poping method, which can save energy for pretreatment and enzyme loading for enzymatic hydrolysis. The chemical analysis of holocellulose of non and popping treated waste wood showed 65.9% and 58.8%, and the lignin, organic extracts and ash were increased by 3%, 4% and 0.7% after pretreatment, respectively. The reducing sugar yields of pretreated waste wood were increased four times more than non-pretreated one and the synergistic effect of cellulase and xylanase were evaluated compare with individual enzyme treatment. Especially, enzyme cocktail (cellulase 50 U and xylanase 50 U) treatment was very efficient in 1% substrate (50 mg). Also, glucose and xylose conversion rate of pretreated waste wood by GC analysis were 45.9% and 38.7%, respectively.
In order to identify probiotic microorganisms, 25 isolates of Lactobacillus sp. were selected from kimchi based on their growth rates, lactic acid production and salt tolerance. The isolate JK-01 was identified as Lactobacillus plantarum by the API kit and 16S rDNA analysis (99.9% of homology), and named as L. plantarum JK-01. The maximum number of L. plantarum JK-01 was reached at 18 hr fermentation in MRS broth and the pH gradually decreased to 4.5. L. plantarum JK-01 showed high enzyme activities for xylanase, amylase, protease, and phytase on MRS agar plates containing each substrate. L. plantarum JK-01 showed high resistance to acidic pH and bile salts, and grew well even at pH 2.0 and 1.0% bile salt. In particular, L. plantarum JK-01 showed high heat stability as shown by $3.3{\times}10^3$ CFU/mL at $60^{\circ}C$. The isolate showed remarkable antimicrobial activity against E. coli in MRS broth based on its disappearance after 18 hr and clear zone formation using a paper disk assay. These results suggest that L. plantarum JK-01 may be probiotic in nature.
Oh, Youn-Lee;Nam, Youn-Keol;Jang, Kab-Yeul;Kong, Won-Sik;Oh, Min ji;Im, Ji-Hoon
Journal of Mushroom
/
v.15
no.3
/
pp.134-138
/
2017
The white button mushroom, Agaricus bisporus, is commercially the fifth most important edible mushroom, accounting for the production of 9,732 tons of mushrooms in Korea in 2015. The genus Agaricus has been known for its potential to degrade lignocellulosic materials. Chemical analyses carried out during the cultivation of A. bisporus indicated that the cellulose, hemicellulose, and lignin fractions were changed preferentially for both vegetative growth and sexual reproduction. We screened A. bisporus strains for effective biodegradation through extracellular enzyme activity using cellulase, xylanase, and ligninolytic enzymes. The enzyme biodegradations were conducted as follows: mycelia of collected strains were incubated in 0.5% CMC-MMP (malt-mops-peptone), 0.5 Xylan-MMP, and 0.5% lignin-MMP media for 14 days. Incubated mycelia were stained with 0.2% trypan blue. Eighteen strains were divided into 8 groups based on different extracellular enzyme activity in MMP media. These strains were then incubated in sterilized compost and compost media for 20 days to identify correlations between mycelial growth in compost media and extracellular enzyme activity. In this study, the coefficient of determination was the highest between mycelial growth in compost media and ligninolytic enzyme activity. It is suggested that comparison with ligninolytic enzyme activity of the tested strains is a simple method of screening for rapid mycelial growth in compost to select good mother strains for the breeding of A. bisporus.
To search probiotic microorganisms, we isolated Lactobacillus sp. from kefir, The Lactobacillus sp. strain showed $99.5\%$ of identity to species Lactobacillus rhamnosus by API kit. Lactobacillus rhamnosus showed high resistances to acidic environment, which grew well even at pH 2.0 and $1.0\%$ bile salt Enzyme activity of Lactobacillus rhamnosus was higher in amylase ($0.673\;{\mu}mol/min/mg$) than that in xylanase ($0.288\;{\mu}mol/min/mg$), cellulase($0.117\;{\mu}mol/min/mg$) and phytase($0.269\;{\mu}mol/min/mg$). Especially, the Lactobacillus rhamnosus showed high heat stability which remained $1{\times}10^6\;CFU/ml$ at $60^{\circ}C$. The maximum numbers of Lactobacillus rhamnosus on growth owe was reached at 24 h fermentation and pH was decreased to 4.6. The resistances of Lactobacillus rhamnosus to acidic pH and bile salt were better than that of Lactobacillus acidophilus used as control. When Lactobacillus rhamnosus was cultured with E. coli in MRS broth, E. coli was disappeared after 18 h. These result suggest that the isolated Lactobacillus rhamnosus has a useful probiotics properties.
To search direct fed microbials, we isolated a Candida sp. from kefir grain. The isolated Candida sp. strain showed 99.8% of identity to the species of Candida kefyr by API 20C kit. Enzyme activity of Candida kefyr was higher in amylase (0.33±1.12μmol/min/mg) than that in phytase (0.052±0.98μmol/ min/mg) cellulase(0.051±μmol/min/mg) and xylanase (0.011±0.98mol/min/mg). The maximum numbers of Candida kefyr in growth curve were reached at 30 h fermentation. Candida kefyr showed high resistances to acidic environment, which was not perfectly extincted even at pH 2.0. And it showed high tolerance to bile salt which had almost 97.2% of survival in the presence of 1.0% bile salt.Especially, Candida kefyr showed high heat stability which remained 10% of initial microorganisms at 60℃. Candida kefyr was not generally inhibited by most of 11 antibiotic agent which contained tetracycline groups. These results suggest that the isolated Candida kefyr has a useful properties as probiotics.
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