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Kafe 바로가기주관연구기관 | 국립농업과학원 National Institute of Agricultural Sciences |
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보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 | 한국어 |
발행년월 | 2014-01 |
과제시작연도 | 2010 |
주관부처 | 농촌진흥청 Rural Development Administration(RDA) |
등록번호 | TRKO201400011014 |
과제고유번호 | 1395019999 |
사업명 | 농업생물연구 |
DB 구축일자 | 2014-07-05 |
DOI | https://doi.org/10.23000/TRKO201400011014 |
Ⅳ. 연구개발결과
□ 제1세부과제: 동애등에 유래 변환산물 이용기술 개발(박관호)
○ 동애등에를 이용한 음식물쓰레기 퇴비화 기술 개발
○ 동애등에 유래 분변토 이용 기술 개발
○ 동애등에를 이용한 사료화 기술 개발
□ 제2세부과제: 동애등에를 이용한 음식물쓰레기 처리 현장적용기술 개발(최영철)
○ 농가현장접목에 따른 대량증식 기술 개발
○ 농가현장의 교미산란장의 규격화 및 산란량 조절
○ 동애등에를 이용한 부산물의 농가 소득원 발굴
□ 제3세부과제: 동애등에 유래 소화효소 특성성분 및
Ⅳ. 연구개발결과
□ 제1세부과제: 동애등에 유래 변환산물 이용기술 개발(박관호)
○ 동애등에를 이용한 음식물쓰레기 퇴비화 기술 개발
○ 동애등에 유래 분변토 이용 기술 개발
○ 동애등에를 이용한 사료화 기술 개발
□ 제2세부과제: 동애등에를 이용한 음식물쓰레기 처리 현장적용기술 개발(최영철)
○ 농가현장접목에 따른 대량증식 기술 개발
○ 농가현장의 교미산란장의 규격화 및 산란량 조절
○ 동애등에를 이용한 부산물의 농가 소득원 발굴
□ 제3세부과제: 동애등에 유래 소화효소 특성성분 및 산업적 이용연구(박관호)
○ 동애등에 유래 효소 분석
○ 소화효소의 분자유전학적 특성 및 소화효소의 발현벡터 구축
○ 소화효소의 활성 분석 동애등에 유래 유용소재 개발 및 이용
□ 제4세부과제: 동애등에 장내 공생미생물 관련 유용유전자 탐색 및 이용(이성재)
○ 단백질 효소 분비능이 뛰어난 Bacillus amyloliquefaciens 특성분석
○ 신 소화효소 분비 세균의 동정을 통한 생물자원 확보(2종)
: Bacillus amyloliquefaciens KBL091, Chryseobacterium sp. nov. KBL621
□ 제5세부과제: 동애등에 유래 신소재 항균활성 물질 개발(전복실)
○ 폐렴 간균에 대한 동애등에 유래 항균 조성물: Adipic acid 등(특허출원)
○ 핵산이산 Hexanedioic acid 체외 in vitro 실험에서 항 폐렴간균(Klebsiella pneumoniae) 효능확인
□ 제6세부과제: 동애등에의 현장적용에 따른 환경 및 생태계 영향 평가(최한수)
○ 동애등에 자연 서식지에서 생태학적 특성 조사(출현 시기, 서식지 특성 등)
○ 동애등에 분변토에 대한 영향 예측 및 저감방안 수립
Black Soldier fly casts produced by eco-friendly treatment of food waste with black soldier fly was composting cast mixed food-waste compost and soldier fly pupa. This study was conducted to evaluate a possibility of SFC as soil amendment by investigating soil properties and zoysiagrass growth. Amen
Black Soldier fly casts produced by eco-friendly treatment of food waste with black soldier fly was composting cast mixed food-waste compost and soldier fly pupa. This study was conducted to evaluate a possibility of SFC as soil amendment by investigating soil properties and zoysiagrass growth. Amendments were selected cocopeat, compost and SFC and their ratio mixed with sand were non-amendment(NA), 7% cocopeat(Coco), 7% compost(Comp), 3.5% soldier fly casts (1/2 SFC) and 7% soldier fly casts(SFC). In soil properties, SFC was increased CEC than NA and T-N, Av-P2O5 and exchangeable K in soil than Coco. Compared with Coco and Comp, turf quality such as turf color index and chlorophyll index in SFC was higher and root length of zoysiagrass longer. Growth of shoot, runner and root in SFC was increase than that of Coco and Comp. These results indicated that 7% SFC applied as soil amendment was improved growth and quality of zoysiagrass by prompting soil productivity and nutrient availability in soil and root growth.
And the other study was conducted to evaluate the feeding value of black soldier fly pupa(BSFP) on the egg production in layer. One hundred twenty laying hens were assigned to 4 treatments (0, 5, 7 and 10% BSFP) with 3 replicates for 8 weeks. The formulae of 4 experimental diets were modified to contain 18% CP and 2,900 kcal/kg. All the treated groups differed in feed intake, egg production, egg weight and egg mass compared to those of a control group. The egg weight and production were increased (p< 0.05) in BSFP groups compared to the control. The HDL-C was increased (p< 0.05) in BSFP groups compared to the control. The level of IgG and Haugh unit were increased (p< 0.05) in BSFP groups compared to the control. These result suggested that the addition of BSFP to the layer diet could improve the egg quality and the laying performance.
Also, the other experiment was conducted to determine the value of black soldier flylarvae and pupae as feed supplements for fresh-water fish (Carassius auratus). Black soldier fly larvae grown on food waste were fed to freshwater fish, live larvae alone, dried larvae, dried pupae and combination with commercial fish diet. A 16-week feed trial was conducted to determine whether black soldier fly larvae could be used as a feed supplement in fresh-water fish. The growth performance indicated that fish fed the live larvae ranked consistently higher, and in combination with commercial fish diet and dried larvae and pupae were next higher. The finding of this study suggest that black soldier fly larvae and pupae may be valuable feedstuff in commercial fish production.
We investigated the biochemical properties of digestive enzymes released from the salivary gland and gut of the black soldier fly. Because the gut extracts of the black soldier fly larvae had high amylase, lipase and protease activities, we suggested that the black soldier fly might belong to the polyphagous insect group. In addition, a strong trypsin-like protease activity was observed in the gut extracts of the black soldier fly larvae. Higher activities of leucine arylamidase, αgalactosidase, βgalactosidase, αJmannosidase and αfucosidase were observed from the gut extracts of the black soldier fly larvae compared with those of house fly larvae. These findings may explain previous reports that the black soldier fly larvae can digest food wastes and organic materials more efficiently than any other known species of fly.
To investigate the molecular scavenging capabilities of the larvae of H . illucens, two serine proteases (SPs) were cloned and characterized. Multiple sequence alignments and phylogenetic tree analysis of the deduced amino acid sequences of Hi-SP1 and Hi-SP2 were suggested that Hi-SP1 may be a chymotrypsin- and Hi-SP2 may be a trypsin-like protease. Hi-SP1 and Hi-SP2 3-D homology models revealed that a catalytic triad, three disulfide bonds, and a substrate-binding pocket were highly conserved, as would be expected of a SP. E. coli expressed Hi-SP1 and Hi-SP2 showed chymotrypsin or trypsin activities, respectively. Hi-SP2 mRNAs were consistently expressed during larval development. In contrast, the expression of Hi-SP1 mRNA fluctuated between feeding and molting stages and disappeared at the pupal stages. These expression pattern differences suggest that Hi-SP1 may be a larval specific chymotrypsin-like protease involved with food digestion, while Hi-SP2 may be a trypsin-like protease with diverse functions at different stages.
The antibacterial effects of larval extract from Hermetia illucens, commonly known as the black soldier fly, was investigated in the present study. Gas chromatography–mass spectrometry analysis identified the active compound within this larval extract as hexanedioic acid. The antibacterial effects of hexanedioic acid were investigated in mice infected with Klebsiella pneumoniae. After administration of hexanedioic acid, infected mice showed decreased lung bacterial loads and lower rates of body weight loss compared to those in the infection-only control group. Based on lung bacterial loads,oral hexanedioic acid treatment showed better protection than intraperitoneal treatment. Administration of hexanedioic acid by either method significantly decreased body weight loss when compared to the control group.
Histopathology confirmed that daily administration of hexanedioic acid for 10 days showed zero toxicity to the kidneys or livers of mice. Therefore, hexanedioic acid may be a novel candidate antibacterial agent.
As it is known that food waste can be reduced by the larvae of Hermetia illucens (Black soldier fly, BSF), the scientific and commercial value of BSF larvae has increased recently.
We hypothesised that the ability of catabolic degradation by BSF larvae might be due to intestinal microorganisms. Herein, we analysed the bacterial communities in the gut of BSF larvae by pyrosequencing of extracting intestinal metagenomic DNA from larvae that had been fed three different diets. The 16S rRNA sequencing results produced 9,737, 9,723, and 5,985 PCR products from larval samples fed food waste, cooked rice, and calf forage, respectively. A BLAST search using the EzTaxon program showed that the bacterial community in the gut of larvae fed three different diets was mainly composed of the four phyla with dissimilar proportions. Although the composition of the bacterial communities depended on the different nutrient sources, the identified bacterial strains in the gut of BSF larvae represented unique bacterial species that were unlike the intestinal microflora of other insects. Thus, our study analysed the structure of the bacterial communities in the gut of BSF larvae after three different feedings and assessed the application of particular bacteria for the efficient degradation of organic compounds.
The Bacillus amyloliquefaciens KBL091 was isolated from intestine of the larvae of Hermetia illucens (Black soldier fly, BSF) which is food waste reducing organism. Strain KBL091 has been identified as a strong decomposer that secretes protease, amylase, and lipase. Especially, the strain KBL091 has strong protease activity much more than other aerobic strains from BSF larval intestine. The strong protease activity started at mid-log phase of culture growth contrast other Bacillus strains which protease activity was enhanced at beginning of stationary phase. However, from secretome analysis by ammonium sulfate precipitation of extracellular culture medium, we not found new additional protease. The protease was purified by additional DEAE-sepharose chromatography and examined for enzymatic characteristics. The enzyme exhibited highest activity at temperature ranging at 45° and pH 6.0 - 9.0. The enzyme activity was 25% enhanced in the presence of 1 mM cobalt. It is the unique property from bacterial
originated proteases.
A novel Gram-negative, facultative anaerobic, non-motile and short rod-shaped bacterium, strain KBL009T,was isolated from the larval gut of Hermetia illucens. Strain KBL009T grew optimally at37°, at pH6.0 and with 1-2% (w/v) NaCl. The 16S rRNA gene sequence of strain KBL009T showed 97.6% similarity to that of Paenalcaligenes hominis CCUG 53761AT indicating its classification with the genus Paenalcaligenes. The major fatty acids were cyclo-C17:0 and C16:0 and summed features 2 (comprising C14:03-OH/iso-C16:1).
The respiratory quinones were ubiquinone-8 (Q-8), predominating and a minor amount of Q-7. The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, one unknown aminolipid and five unknown polar lipids. The polyamine pattern contained predominantly putrescine and relatively high amounts of spermidine. The betaproteobacteria-specific 2-hydroxyputrescine could only be detected in trace amounts.
The G+C content of genomic DNA was 56.1 mol%. Results from DNA-DNA hybridization with Paenalcaligenes hominis KCTC 23583T unambiguously demonstrated that strain KBL009T represents a novel species in the genus Paenalcaligenes. Based on phenotypic, genotypic and phylogenetic characterization, the novel species Paenalcaligenes hermetiae sp. nov. is proposed. The type strain is KBL009T(=KACC16840T=JCM18423T).An emended description of the genus Paenalcaligenes is also provided.
A novel, Gram-negative, facultative anaerobic, motile and short rod-shaped bacterium, strain KBL006T was isolated from the larval gut of Hermetia illucens, Black soldier fly. The 16S rRNA gene sequence of strain KBL006T showed 96.4% similarity to that of Wohlfahrtiimonas chitiniclastica S5T. Strain KBL006T grew optimally at 30°, at pH8.0 and in the presence of 1-2% (w/v) NaCl. Oxidase activity and catalase activity were positive.
The major fatty acids were C18:1ωc, C14:0, and C16:0. The major respiratory quinone was ubiquinone-8(Q-8).The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol, and two phospholipids. The G+C content of genomic DNA was 45.2 mol%. Based on these polyphasic data, for which the name Wohlfahrtiimonas larvae sp. nov. is proposed. The type strain is KBL006T(=KACC16839T=JCM18424T).
In this study, the ecological characteristics and the genetic characteristics of H . illucens collected in Korea and potential usage of the compost were conducted. H . illucens was confirmed to be less affected in effects of latitude, longitude, altitude. H. illucens CO1 specific primer set (BSFCO1-f: CAA CAA ATC ATA AAG ATA TTG GAA C BSFCO1-r: GTC CGA AGA ATC AAA ATA AAT GYT G) was used and this set worked very efficiently throughout the gradient range from 44 ˚ to 55 ˚. There were 9 CO1 haplotypes and the parsimony informative sites were 39 nucleotide sites. Among them haplotype 9 was most common (108 individuals, 52.4%), and it appeared at every collection site except Seoul, Danyang and Buyeo.Seven out of 9 haplotypes were found in South-central area based on biological floral region and consistently the rate of point mutations such as transition and transversion was relatively high. Genetic molecular diversity was high in each region at 35~40%. However, it was relatively low in the South-central region. This indicates each individual collected in the area was genetically similar. Chemical analysis of H. illucens compost was suitable as a fertilizer. When Phaseolus vulgaris is raised in culture soil treated by H. illucens compost, culture soil acted as a promoting factor of growth of leaf area, total dry weight, root dry weight, stem dry weight, and leaf dry weight. Treated by H. illucens compost, the relative growth rate of Phaseolus vulgaris acted as an increasing factor of relative growth rate from the beginning of growth and also had a great effect on the relative growth rate until the late stage of growth. There is highly significance statistically between NAR and RGR, but no significance statistically between RGR and LAR of treatment groups of H . illucens compost. In addition, the treatment of H. illucens compost acted as a factor of increasing the thickness of leaves from the beginning of growth and the tendency of thickening leaves from the beginning of growth continued the effect through the late period of
growth. Mixed with 50% of H. illucens compost rather than 25%, it showed a greater effect of promoting growth.
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