• 한국초지조사료학회지
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• 제28권3호
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• pp.229-236
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• 2008

본 시험은 사료용 총체 벼 사일리지 전용 미생물 첨가제 개발을 위하여 2003년부터 2005년까지 축산연구소에서 수행되었다. 새로운 총체 벼 전용 첨가제 개발을 위해 수집된 28점의 총체 벼 사일리지로부터 미생물을 선발하여 전체 5개 균주(R4-1, R7-1, R7-2, R10-1 및 R12-1)를 분리하였다. 분리된 균주중 4종(R4-1, R7-1, R7-2 및 R10-1)은 Lactobacillus plantarum으로 동정되었고 R12-1은 L. pentosus로 동정되었다. 황숙기에 수확된 총체 벼는 사일리지를 제조할때 선발된 5개 균주 이외에 시중에서 판매되는 첨가제 3종을 접종을 하여 60일간 실온에서 저장한 후 평가한 결과는 다음과 같다. 저장된 사일리지의 pH와 초산 함량은 첨가제를 처리한 구에서 낮았고 젖산 함량은 높았다(p<0.05). 특히 R7-1은 초산 함량이 가장 낮았고 젖산 함량이 가장 높았다. 조단백질 함량은 R7-2 첨가제 처리구에서 가장 높았으며 ADF 및 NDF 함량은 R7-1에서 가장 낮았다(p<0.05). 선발된 균주들은 대부분 사일리지 품질개선효과를 나타내었으나 R7-1이 그 중에서 가장 효과적이었다. 본 미생물은 한국농용미생물 협회에 NLRI 401로 등록되었다.

• Animal Bioscience
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• 제35권9호
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• pp.1379-1389
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• 2022

Objective: This study identified the major lactic acid bacteria (LAB) strains from different fermented total mixed rations (FTMRs) via metataxonomic analysis and evaluated the ability of their standard strain as ensiling inoculants for corn stover silage. Methods: The bacterial composition of eight FTMRs were analyzed by 16S rDNA sequencing. Corn stover was ensiled without LAB inoculation (control) or with 1×10⁶ cfu/g LAB standard strain (Lactobacillus vaginalis, Lactobacillus reuteri, Lactobacillus helveticus, or Lactobacillus paralimentarius) selected from the FTMRs or 10 g/t commercial silage inoculant (CSI) around 25℃ for 56 days. For each inoculation, a portion of the silage was sampled to analyze ensiling characteristics at time intervals of 0, 1, 3, 7, 14, 28, and 56 days, gas production (GP), microbial crude protein and volatile fatty acids as the measurements of rumen fermentation characteristics were evaluated in vitro with the silages of 56 days after 72 h incubation. Results: Lactobacillus covered >85% relative abundance of all FTMRs, in which L. pontis, L. vaginalis, L. reuteri, L. helveticus, and L. paralimentarius showed >4% in specific FTMRs. CSI, L. helveticus, and L. paralimentarius accelerated the decline of silage pH. Silage inoculated with L. paralimentarius and CSI produced more lactic acid the early 14 days. Silage inoculated with L. paralimentarius produced less acetic acid and butyric acid. For the in vitro rumen fermentation, silage inoculated with CSI produced more potential GP, isobutyric acid, and isovaleric acid; silage inoculated with L. helveticus produced more potential GP and isovaleric acid, silage inoculated with L. paralimentarius or L. reuteri produced more potential GP only. Conclusion: The standard strain L. paralimentarius (DSM 13238) is a promising ensiling inoculant for corn stover silage. The findings provide clues on strategies to select LAB to improve the quality of silage.

• 생명과학회지
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• 제17권12호
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• pp.1682-1688
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• 2007

다기능성 농업용 미생물 제제를 개발하기 위하여 생물방제 및 생물비료 활성을 가지는 미생물을 탐색하였다. 본 연구실에서 분리 및 동정된 균주가운데 Pantoea agglomerans 및 Bacillus megaterium을 실험군주로 선정하였으며, 경남 밀양에 위치하는 양계장 부근 부엽토로부터 새로운 다목적 세균 MF12를 분리하였다. 형태학적, 배양적, 생화학적 특성 및 16S rDNA 염기서열을 분석한 결과, MF12는 Bacillus pumilis로 동정되었다. 이 균주들의 불용성 인산 가용능, IAA 및 siderophore 생성능, ammonification ability, 식물병원성 진균 세포성분 분해효소 생성능 및 항진균능을 조사하였다. P. agglomerans는 고체배지에서 불용성 인산을 가용화할 수 없었으나 액체배지에서는 가용성 인산을 생성하였다. 상기 모든 균주들은 배양시간에 따라 $3{\sim}639{\mu}g/ml$의 IAA를 생성하였으며, P. agglomerans만이 siderophore를 생성하였다. 이 균주는 pectinase와 lipase를 생성하였다. B. megaterium은 amylase, pretense 및 lipase를 생성한 반면 B. pumilisr는 protease와 lipase를 생성하였다. P. agglomerans는 Fusarium oxysporum과 Colletotrichum gloeosporioides의 생육을 억제하였으며, B. pumilis는 Botrytis cinerea, Sclerotinia sclerotiorum 및 Phythium ultimum의 생육을 억제하였다.

• Animal Bioscience
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• 제34권4호
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• pp.642-651
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• 2021

Objective: This study aimed to determine the effects of different roughages in total mixed ration (TMR) inoculated with or without coculture of Lactobacillus acidophilus (L. acidophilus) and Bacillus subtilis (B. subtilis) on in vitro rumen fermentation and microbial population. Methods: Three TMRs formulations composed of different forages were used and each TMR was grouped into two treatments: non-fermented TMR and fermented TMR (F-TMR) (inoculated with coculture of L. acidophilus and B. subtilis). After fermentation, the fermentation, chemical and microbial profile of the TMRs were determined. The treatments were used for in vitro rumen fermentation to determine total gas production, pH, ammonianitrogen (NH3-N), and volatile fatty acids (VFA). Microbial populations were determined by quantitative real-time polymerase chain reaction (PCR). All data were analyzed as a 3×2 factorial arrangement design using the MIXED procedure of Statistical Analysis Systems. Results: Changes in the fermentation (pH, lactate, acetate, propionate, and NH3-N) and chemical composition (moisture, crude protein, crude fiber, and ash) were observed. For in vitro rumen fermentation, lower rumen pH, higher acetate, propionate, and total VFA content were observed in the F-TMR group after 24 h incubation (p<0.05). F-TMR group had higher acetate concentration compared with the non-fermented group. Total VFA was highest (p<0.05) in F-TMR containing combined forage of domestic and imported source (F-CF) and F-TMR containing Italian ryegrass silage and corn silage (F-IRS-CS) than that of TMR diet containing oat, timothy, and alfalfa hay. The microbial population was not affected by the different TMR diets. Conclusion: The use of Italian ryegrass silage and corn silage, as well as the inoculation of coculture of L. acidophilus and B. subtilis, in the TMR caused changes in the pH, lactate and acetate concentrations, and chemical composition of experimental diets. In addition, F-TMR composed with Italian ryegrass silage and corn silage altered ruminal pH and VFA concentrations during in vitro rumen fermentation experiment.

• Asian-Australasian Journal of Animal Sciences
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• 제33권8호
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• pp.1252-1264
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• 2020

Objective: To effectively utilize crop by-product resources to address the shortage of animal feed during the dry season in Africa, the community of natural lactic acid bacteria (LAB) of corn stover and sugarcane tops and fermentation characteristics of silage were studied in Mozambique. Methods: Corn stover and sugarcane tops were obtained from agricultural field in Mozambique. Silage was prepared with LAB inoculant and cellulase enzyme and their fermentation quality and microbial population were analyzed. Results: Aerobic bacteria were the dominant population with 10⁷ colony-forming unit/g of fresh matter in both crops prior to ensiling, while 10⁴ to 10⁷ LAB became the dominant bacteria during ensiling. Lactobacillus plantarum was more than 76.30% of total isolates which dominated silage fermentation in the LAB-treated sugarcane top silages or all corn stover silages. Fresh corn stover and sugarcane tops contain 65.05% to 76.10% neutral detergent fiber (NDF) and 6.52% to 6.77% crude protein (CP) on a dry matter basis, and these nutrients did not change greatly during ensiling. Corn stover exhibits higher LAB counts and water-soluble carbohydrates content than sugarcane top, which are naturally suited for ensiling. Meanwhile, sugarcane tops require LAB or cellulase additives for high quality of silage making. Conclusion: This study confirms that both crop by-products contain certain nutrients of CP and NDF that could be well-preserved in silage, and that they are potential roughage resources that could cover livestock feed shortages during the dry season in Africa.

• 한국지하수토양환경학회지:지하수토양환경
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• 제17권3호
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• pp.32-38
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• 2012

This study was conducted to evaluate the slurping process affecting the variation of free product and VOCs concentration and the bioaugmentation effect on bioremediation process. Free products and soil gas were extracted from 30 extraction wells installed in a petroleum contaminated area. The extraction system was operated for 10 hours per day with 1 hour on-and-off mode. The thickness of free product in extraction well was decreased from 11.7 cm to 4.5 cm and the VOCs concentration was increased from 10.37 ppm to 30.78 ppm during the operation period. After the slurping process for 2 months, contaminated soil was treated with bioremediation process in 2 cells, $15{\times}40$ m, biologically enhanced with adjusting oxygen, moisture and nutrients concentration. Total 1,400 L of microbial inoculant, Naturesys. (Dong Myung Ent. Co.) was added to the pile B, which has an outstanding ability for degrading petroleum hydrocarbons. The results showed that bioremediaton effect in soil with the microorganisms solution is 33% higher than that in soil with only residual bacteria.

• Journal of Microbiology and Biotechnology
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• 제19권11호
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• pp.1431-1438
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• 2009

The role of plant growth-promoting rhizobacteria (PGPR) in the phytoremediation of heavy-metal-contaminated soils is important in overcoming its limitations for field application. A plant growth-promoting rhizobacterium, Serratia sp. SY5, was isolated from the rhizoplane of barnyard grass (Echinochloa crus-galli) grown in petroleum and heavy-metal-contaminated soil. This isolate has shown capacities for indole acetic acid production and siderophores synthesis. Compared with a non-inoculated control, the radicular root growth of Zea mays seedlings inoculated with SY5 can be increased by 27- or 15.4-fold in the presence of 15 mg-Cd/l or 15 mg-Cu/l, respectively. The results from hydroponic cultures showed that inoculation of Serratia sp. SY5 had a favorable influence on the initial shoot growth and biomass of Zea mays under noncontaminated conditions. However, under Cd-contaminated conditions, the inoculation of SY5 significantly increased the root biomass of Zea mays. These results indicate that Serratia sp. SY5 can serve as a promising microbial inoculant for increased plant growth in heavy-metal-contaminated soils to improve the phytoremediation efficiency.

• 한국가금학회지
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• 제32권4호
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• pp.291-300
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• 2005

남은 음식물에서 Aspergillus oryzae(AO)의 유용한 배양조건(시간, 수분함량, 접종균수)을 구하기 위하여 2가지 실험과 닭에서 그 배양물의 영양소 이용율을 구명하기 위하여 본 시험을 수행하였다. <시험 1> 적정 배양시간과 접종균수의 측정을 위하여 수분함량을 30, 40, 50, $60\%$로 정하였다. 잔반 사료에 접종하는 seed로서 1mL Aspergillus oryzae(AO) $(1.33\times10^5\;CFU/mL)$를 이용하였다. 배양을 위한 적정 수분함량은 $40\~50\%$였으며 배양시간은 72시간 이상이었다. 계속하여 AO 균수에 의한 효과를 결정하기 위해 AO seed 0.25, 0.50, 0.75, 1.00mL를 남은음식물 사료에 접종하였다. AFW의 AO균수를 비교하기 위하여 72시간과 96시간 배양하였으나, 접종량에 따른 유의차는 없었다. 다시 수준을 낮추어 0.01, 0.05, 0.10mL를 접종하고 72시간과 96시간동안 배양한 결과, 72시간 배양 AO colony 수는 0.1 mL 접종 처리 구에서 가장 많았다. <시험 2> FW의 배양조건을 결정한 후에, 20수의 5주령 Hubbard종을 공시하여 소화율 시험을 수행하였다. 처리구는 T1-대조구, T2-기초사료 $60\%$+건조잔반 $40\%$, T3-기초사료 $60\%$+건조잔반 $20\%$+AO균 접종 건조잔반 $20\%$, T-4-기초사료 $60\%$+AO균 접종 건조잔반 $40\%$ 처리구 등 총 4처리구, 처리구당 5반복씩 20수를 인공항문 장착하여 소화율을 조사하였다. 영양소 소화율에 있어 일반성분과 아미노산은 대조구에 비해 T2 처리구가 전성분에서 낮았고(p<0.05), AFW(T3, T4)처리구에 비해 조지방과 인의 소화율에서 차이를 보였다(p<0.05). T3와 T4는 T2에 비해 조섬유 및 조회분의 소화율이 증가하였다(p<0.05). 대조구는 T3와 T4에 비해 필수아미노산인 arginine, leucine, phenylalanine의 소화율이 높았으나(p<0.05), T3와 T4 처리구는 T2 처리구에 비해 필수아미노산 arginine, lysine 및 thereonine과 비필수아미노산 alanine의 소화율이 개선되었다(p<0.05). 이러한 결과들로부터 건조 잔반의 AO 배양조건 구명과 영양소 이용율 시험을 통해 FW사료의 품질이 기초사료에 비해 영양소 이용율이 낮고, AFW 사료가 AO에 의해 개선될 수 있음을 시사한다

• 한국환경농학회지
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• 제17권3호
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• pp.254-259
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• 1998

농가에서 실행되고 있는 유기농업의 실증적 연구를 위하여 돈분, 또는 돈분과 계분을 혼합하여 만든 발효퇴비를 10a 당 3, 6, 12ton씩 시비하고 다시 미생물제를 처리한 후 토마토의 생육, 수량 및 토양 중 양분의 변화를 조사하였던 바 다음과 같은 결과를 얻었다. 1. 토마토 초장은 퇴비 시용구에서 관행구에 비하여 길었으며 엽폭은 관행구에서 더 긴 것으로 나타났다. 엽록소 함량은 관행구에서 계속적으로 높은 값을 보였으며 퇴비 시용량의 차이에 따른 엽록소 함량의 차이는 정식 45일 후부터 아주 낮아지는 것으로 나타났다. 2. 미생물제를 사용하여 충분히 발효시킨 경우 12ton/10a의 시용에서 토마토의 수량이 관행구의 50-60%에 불과하였으나, 재배도중 추비하였을 경우에는 6ton/10a 시용구에서 관행구의 82%정도에 달하는 수량을 얻었다. 3. 일차 발효된 퇴비의 시용 후에 미생물제의 토양 살포는 질소성분의 빠른 유실을 초래하였으며 토양중 미생물의 밀도를 상당히 높혀주는 것으로 나타났다. 4. 과실의 색깔은 모든 퇴비 시용구에서 관행구에 비하여 붉은 색이 선명하게 나타났으며, 당도와 산도는 퇴비 시용량이 많은 12ton/10a 처리구에서 가장 높게 나타났으며, 경도는 관행구에서 유기물 시용구에 비하여 높게 나타났다. 5. 퇴비 시용구에서 $NO_3-N$와 $NH_4-N$의 함량은 시용 직후에는 많은 양이 검출되었으나 급속히 감속되어 4-5주 후부터 아주 미량이 검출되었다. 그러나 인산, 칼륨, 마그네슘 및 칼슘의 함량은 서서히 감소되는 경향을 보였다.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.711-717
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• 2008

To speed up the conversion of rice straw into feeds in a low-temperature region, a start culture used for ensiling rice straw at low temperature was selected by continuous enrichment cultivation. During the selection, the microbial source for enrichment was rice straw and soil from two places in Northeast China. Lab-scale rice straw fermentation at $10^{\circ}C$ verified, compared with the commercial inoculant, that the selected start culture lowered the pH of the fermented rice straw more rapidly and produced more lactic acid. The results from denatured gradient gel eletrophoresis showed that the selected start culture could colonize into the rice straw fermentation system. To analyze the composition of the culture, a 16S rRNA gene clone library was constructed. Sequencing results showed that the culture mainly consisted of two bacterial species. One (A) belonged to Lactobacillus and another (B) belonged to Leuconostoc. To make clear the roles of composition microbes in the fermented system, quantitative PCR was used. For species A, the DNA mass increased continuously until sixteen days of the fermentation, which occupied 65%. For species B, the DNA mass amounted to 5.5% at six days of the fermentation, which was the maximum relative value during the fermentation. To the authors' best knowledge, this is the first report on ensiling rice straw with a selected starter at low temperature and investigation of the fermented characteristics.