Korean Journal of Microbiology (2013) Vol. 49, No. 4, pp. 343-352 DOI http://dx.doi.org/10.7845/kjm.2013.3043 Copyright c 2013, The Microbiological Society of Korea 보문 매몰지조기안정화를위한유용미생물의효과에관한연구 김현숙 1 박수정 2 정원화 2 Sathiyaraj Srinivasan 3 이상섭 3 * 1 경기대학교생명공학과, 2 국립환경과학원, 3 경기대학교생명과학과 The Study on the Effect of Efficient Microorganism for Early Stabilization of the Burial Sites Hyun-Sook Kim 1, Sujung Park 2, Weonhwa Jheong 2, Sathiyaraj Srinivasan 3, and Sang-Seob Lee 3 * 1 Department of Bioengineering, Kyonggi University, Suwon 443-760, Republic of Korea 2 Environmental Infrastructure Research Department, Water Supply & Sewerage Research Division, National Institute of Environmental Research, Incheon 404-708, Republic of Korea 3 Department of Life Science, Kyonggi University, Suwon 443-760, Republic of Korea (Received June 26, 2013 / Accepted November 11, 2013) In this study, we have evaluated the effect of efficient microorganisms on odor-removal efficiency and early stabilization of the burial sites. We have developed an efficient microorganism designated as KEM which have the ability to degrade organic compounds and remove odor effectively. Other efficient microorganisms already used on site, such as EM and Bacillus sp., were also compared. We preceded these experiment using lab-scale reactors under three conditions (control, only media and only body) and comparing the effect of with or without the application of tree efficient microorganisms separately. Analysis was focused on eight components (ammonia, TMA, H 2S, methyl mercaptan, dimethyl sulfide, dimethyl disulfide, CO 2 and CH 4), and as a result, efficient microorganisms were shown efficiency in the removal of ammonia and methyl mercaptan. The applied KEM decayed up to 71.2% of the buried meat. We were unable to observe significant differences in microbial communities between efficient microorganisms-treated and non-treated reactors due to the large presence of microorganisms in both soil and carcasses. However, it was possible to observe the effect on odor control and decay rate through the application of efficient microorganisms. Keywords: efficient microorganism, FMD (Foot and Mouth Disease), KEM, odor removing efficiency 최근발생한구제역과조류인플루엔자 (AI) 로인해전국약 4,600개소의매몰지에돼지 213만마리, 소 14만마리, 조류 380 만마리의동물사체를매몰하였고, 그에따른 2차환경오염이우려되고있다 (McBean and Ro Farquhar, 1995; Banks and Wang, 1999; Higgins et al., 2002; Fuller and Manning, 2004). 구제역발생 3년뒤, 지금까지도매몰인근지역은악취와침출수의발생이이어지고있다 (Oberbremer et al., 1990; Angelidaki and Ahring, 1992; Castaldi and Ford, 1992; Cassidy and Hudak, 2001; Wang and Vipulanander, 2001; Hanyang University, 2002; Fava et al., 2004; Ireri et al., 2008). 국민들의생활수준과환경에대한관심이증가함에따라생활환경문제에대한관심과민원역시증가하고있다. 현재안정화효과가검증되지않은유용미생물제제들이매몰지에무분 *For correspondence. E-mail: sslee@kyonggi.ac.kr; Tel.: +82-31-249-9642; Fax: +82-31-245-8868 별하게사용되고있으며 (Ministry of Environment, 2011), 이에대한기본연구가되어있지않은것이실정이다. 따라서본연구에서는랩스케일의반응기를통하여매몰지조기안정화및악취제어에대한유용미생물의효과를알아보려한다 (Pinelli et al., 1997; Zeev and Aharon, 2000; Zhang et al., 2000; Salminen and Rintala, 2001; Janikowski et al., 2002). 재료및방법미생물제제선정및대량배양유용미생물제제로는혼합균주 KEM을개발하였다. KEM은 닉시안과환경미생물은행에서분양받았으며유기물및질소분해능이있는홍색비황혼합광합성박테리아와질소및황화합물악취제어능이뛰어난균주와발효균주를혼합하여만들었다. 또한대조군으로써 EM과바실러스를선정하였으며, 현장에서가장많이사용되고있는유용미생물제제로알려져있다
344 Kim et al. Table 1. The list of efficient microorganism Efficient microorganism KEM EM Bacillus sp. Microorganism Rhodopseudomonas capsulate, Rhodococcus sp., Achromobacter sp., Alcaligenes sp., Paracoccus sp., Thiobacillus sp., Fermented mixture Photosynthetic bacteria, Pichia deserticola, Lactobacillus sp. Bacillus alcalophylus (Table 1). EM과바실러스의배양을위해멸균한 nutrient broth (NB, Difco, USA) 배지 1 L에 1% 계대배양하였다. 바실러스는알칼리조건에서최적배양의조건을보이는것으로확인되어 ph 8.0 으로조정하여배양하였다. EM은통성혐기조건에서, 바실러스는호기조건에서 24 48시간동안배양하였고, 이후대량배양을위해 20 L의배양통에계대하여추가로 3 5일간더배양하였으며모든배양은 28 에서하였다. KEM의증식배양을위해광합성박테리아는 27M media (yeast extract 1.0 g, ethanol 0.5 ml, di-sodium succinate 1.0 g, 0.5% ferric citrate solution 1 ml, KH 2PO 4 0.5 g, MgSO 4 7H 2O 0.4 g, NaCl 0.4 g, NH 4Cl 0.4 g, CaCl 2 2H 2O 0.05 g, 1X Trace element solution SL-6 1 ml), 질소화합물악취제거균주는 ATCC 412 media (NH 4Cl 5.0 g, KH 2PO 4 2.0 g, NaCl 0.5 g, MgSO 4 7H 2O 0.2 g, FeSO 4 0.002 g, MnCl 2 0.002 g, yeast extract 0.2 g), 황화합물악취제거균주는 Thiobacillus selective media (yeast extract 2.0 g, Na 2S 2O 3 5.0 g, KH 2PO 4 1.5 g, Na 2HPO 4 4.5 g, MgSO 4 7H 2O 0.1 g, NH 4Cl 0.3 g), 발효균주는 NB를사용하였다. 각미생물은배지 1 L에 1%(v/v) 을계대배양하였다. 광합성박테리아와발효균주는통성혐기조건으로 48 72시간동안배양하였고, 혼합균주와질소및황화합물악취제거균주는호기조건으로배양하였으며 24 48시간동안배양하였다. 이후대량배양을위해 20 L의배양통에계대하여 3 5일간동안 28 에서배양하였다. 균주는초기주입후, 200일이경과한뒤로는한달에한번씩미생물제제 (1 g/l, w/v) 와수분 500 ml을추가주입해주었다. Fig. 1. Diagram of reactor and lab-scale of reactor. Table 2. The condition of analysis for sulfur compounds Temp. ( ) Temp. ( ) Column DB-dioxin Oven 200 Inlet 280 Detector 300 Table 3. The condition of analysis for CO 2 and CH 4 Column Eluent Flow Rate Detector Column HP-plot Q Oven 23 Inlet 280 Detector 300 Table 4. The condition of analysis for ammonia Table 5. The condition of analysis for TMA Temp. ( ) Waters IC-Pak Cation M/D 0.1 mm EDTA / 3.0 mm Nitric Acid 1 ml/min Conductivity Column HP-5 Oven 220 Inlet 280 Detector 270 가축사체선정및반응기제작가축사체는부분사체를사용하였으며, 실제가축의육질과내장 (3:2) 비율, 혈액의비율 ( 사체무게의 5%) 을고려하여매몰하였다. 반응기는매몰지의축소형모형 (1/35비율로축소 ) 으로제작하였으며, 반응기의상층부에는발생되는가스를포집할수있도록 tedlar bag을연결하였고, 하층부에는발생되는침출수를모을수있도록관을연결하였다 (Fig. 1). 유용미생물적용군에대한대조군으로는미생물을적용하지않은경우 (control), 배지만적용한경우 (only media), 토양에매몰하지않고사체만적용한경우 (only body) 로나누어준비하였다. 가스성분분석배출되는가스성분은가스포집장치와가스를액상으로포집하는장치, 두가지방법으로포집후분석하였다. 기체포집장치와 tedlar bag을사용하여기체를포집한후, 황화합물악취성분인황화수소, 메틸머캅탄 (methyl mercaptane), DMS (dimethyl sulfide), DMDS (dimethyl disulfide) 및탄화수소계성분 (CO 2, CH 4) 은농축시킨후 GC/MS (Gas Chromatography/Mass Spectroscopy) 로분석하였다 (Tables 2 and 3). 질소화합물악취성분인암모니아, TMA (trimethylamine) 은펌프와 impringer 장치를이용하여액상흡수액에포집하였으며, 암모니아는 IC (Ion Chromatography) 로분석하였고 (Table 4), TMA는 GC-FID (Gas Chromatography- Flame Ionization Detector) 로분석하였다 (Table 5). 미생물의군집분포도변화분석미생물의군집분포의변화를분석하기위하여다양한분자
유용미생물의매몰지조기안정화효과 345 Table 6. Molecular methods for microbial community analysis Method Category Format of result Database construction Cost DGGE (denaturing gradient gel electrophoresis) Fingerprint Gel/band image No Low TGGE (temperature gradient gel electrophoresis) Fingerprint Gel/band image No Low SSCP (single strand conformation polymorphism) Fingerprint Gel/band image No Low Clone library Sequencing Sequences Yes High Pyrosequencing Sequencing Sequences Yes Medium 생물학적방법이개발되었다. 대부분의경우 16S rdna를 PCR 로증폭해서이를분석에사용하며 (Table 6), 본연구에서는 DGGE (Denaturing gradient gel electrophoresis) 와 pyro-sequencing 을통해분석하였다 (Amann et al., 1995). Pyrosequencing은 16S rrna 유전자의 V1~ 3 부분을분석하였으며, primer는 V1-9F (5 -X-ACGAGTTTG TCMTGGCTCAG-3 ) 와 V3-541R (5 -X-ACWTTACCGCGG TGCTGG-3 ) 을사용하였다 (Chun et al., 2010). 각시료에서 FastDNA spin kit for soil of fecal (MP Biomedicals, USA) 을이용하여 DNA를추출하였고, 천랩연구소에 Pyrosequencing 분석을의뢰하였다. 또한, DGGE의미생물군집분포도를확인하기위하여, 추출한 DNA를미생물의일반적인 primer 341F (5 -CCTACGGGAGGCAGCAG-3 ) 와 907R (5 -CCGTCAATTC TTTGAGTTT-3 ) 를이용하여 1차 PCR을수행하였다. 그후, GC-clamp가달린 341F (5 -CGCCCGGGG CGCGCCCCGGG CGGGGCCCTAC GGGAGGCAGCA-3 ) 와 907R를이용하여 2차 PCR을수행하였다. 2차 PCR 결과물로 DGGE를수행하였다. DGGE의변성제농도는각 30% 와 60% 을사용하였다. 미생물을적용하지않은경우 5.72 g/day이었고, 유용미생물 EM, KEM, 바실러스를적용하였을경우각각 6.28, 6.20, 5.47 g/day인것으로나타났다. 또한배지만적용하였을경우에는 5.70, 사체만적용한반응기에서는 3.26 g/day이었다. 반응기내의남은가축사체의양은적은량의지방질과소량의부분사체만이관찰되었다. 가축사체분해율에서는유용미생물에따른효과가크게나타나지않았는데이는반응기에함께매몰한토양에존재하는토양미생물에의한작용으로사료된다. 또한가축사체제거율은사체만적용한반응기를제외하고모두약 30% 를보였으며, 유용미생물 EM과 KEM을적용한반응기에서 40, 45.8% 의제거율을보였다. 주기적인수분공급후에는제거율이 67% 까지증가하였으며, EM과 KEM을적용하였을경우 76, 71.2% 까지증가한것을확인하였다. 결과및고찰 다양한조건의모형매몰지의가축사체분해율조사연구가축사체분해속도를간접적으로확인하기위해반응기의전제무게를 2주에한번씩측정하였다. 200일후부터주기적으로수분및미생물제제를공급해준결과확연하게사체분해속도가빨라진것을확인할수있었다 (Fig. 2). 사체분해속도는 Fig. 2. Average weight decreasing rate of in vessel meat. ( ) Control, ( ) Only media, ( ) Only body, (X) EM, (Ж) KEM, and ( ) Bacillus Fig. 3. Tendency of CO 2 emission (A) and CH 4 emission (B). CO 2 was generated continuously with high concentration, CH 4 emission level was increased after 200 days. ( ) Control, ( ) Only media, ( ) Only body, (X) EM, (Ж) KEM, and ( ) Bacillus
346 Kim et al. 탄화수소계성분분석 CO 2 : CO 2 는연구기간동안지속적으로발생되었으며, 사체부패정도에따라배출량이증가되는것을확인하였다 (Fig. 3A). CO 2 의배출잔량은사체만적용한반응기에서 14344.60 mg으로가장높은값을보였고, 사체적용반응기를제외한다른모든반응기에서는모두비슷한값을가지는것으로확인되었다 (Fig. 4A). CH 4 : 초기에발생되지않았으나, 주기적으로수분과미생물제제를주입해준이후부터발생량은점차증가후다시감소되는것을확인하였다 (Fig. 3B). 바실러스를적용하였을경우 27.10 mg으로배출잔량이다른유용미생물을적용한반응기의배출잔량보다가장낮은값을나타내었다. 그러나대조군중, 미생물을적용하지않은반응기와의배출잔량을비교하였을때, 유용미생물을적용한반응기에서의배출잔량이더높은것으로나타났다 (Fig. 4B). 질소화합물악취성분분석암모니아 : 암모니아의발생량은시간이경과될수록전체적으로크게증가하였으며, 균과배지의추가주입에따라발생량의증감추세가모든반응기에서관찰되었다. 초기에모든반응기에서암모니아의발생농도는모두비슷하였고, 200일이지난후에 3개의대조군 ( 미생물을적용하지않은경우, 배지만적용한경우, 사체만적용한경우 ) 의반응기에서많은양의암모니아를발생하였다. 또한, 미생물을적용하지않은반응기에서발생된암모니아의총량이가장많았다 (Fig. 5A). EM, KEM, 바실러스를각각적용하였을경우, 배출잔량은 0.26, 0.33, 0.38 mg으 로나타났다 (Fig. 6A). TMA: TMA는사체가부패될때발생되는악취이다. 초기모든반응기에서사체의분해가활발히일어나며높은농도의 TMA가검출되었으나, 시간이경과할수록발생양은감소하는것을확인하였다 (Fig. 5B). KEM을적용하였을경우 3.45 mg의 TMA 배출잔량을확인하였지만, 다른유용미생물을적용한경우에는 EM과바실러스적용한반응기에서각각 7.53, 7.23 mg 으로대조군반응기보다많은양의 TMA가잔존하고있음을확인하였다 (Fig. 6B). 황화합물악취성분분석황화수소 : 황화수소는아미노산중황을함유하고있는시스테인과메티오닌이분해될때생성된다 (Eweis et al., 1998). 초기한달동안은사체분해가활발히일어나면서많은양의황화수소를발생하였지만, 그이후급격히발생량이감소하여약 40 일이후부터는거의발생되지않는것으로확인되었다 (Fig. 7A). KEM을적용한반응기에서황화수소의잔존량은 0.37 mg으로가장낮은값을보였으나, 사체만매몰한경우를제외한다른대조군반응기에서도각각 0.44, 0.64 mg으로낮은잔존량이확인되었다 (Fig. 8A). Fig. 4. According to decomposed rate of carcass, remained the CO 2 level (A) and CH 4 level (B). Efficient microorganism cannot show the removing efficiency for CO 2 and CH 4. Fig. 5. Tendency of ammonia emission (A) and TMA emission (B). At initial time ammonia emission level very low, however after 200 days emission level increased all vessels. At initial time TMA emission level very high all vessels, but according to time emission level decreased. ( ) Control, ( ) Only media, ( ) Only body, (X) EM, (Ж) KEM, and ( ) Bacillus
유용미생물의매몰지조기안정화효과 347 메틸머캅탄 : 메틸머캅탄은연구기간동안지속적으로발생하는추이를보였으며, 균주의추가주입을하였을때발생량이증가되었다감소되는현상을보였다 (Fig. 7B). 또한, 유용미생물을적용한반응기에서대조군에비해발생량이높은것으로나타났다. 이는주입한미생물에의해사체분해가진행되면서가스가발생되었다가, 이를다시균이제거함으로써가스의발생량이줄어든것으로사료된다. 메틸머캅탄의배출잔량은유용미생물 EM, KEM, 바실러스를적용하였을때각각 0.90, 0.61, 1.41 mg으로나타났다 (Fig. 8B). DMS: DMS는미생물에의해사체분해가일어나, 발생된황화수소의메틸화를통해발생하게되며, 이는황화수소의발생량이줄어드는시점에, DMS의발생량은반대로증가하는것을통해알수있다. 약한달후부터모든조건에서 DMS의발생이측정되었으며바실러스를적용한반응기에서는높은농도의 DMS를발생하였다 (Fig. 7C). 유용미생물을적용한반응기에서의배출잔량은각각 EM은 0.34 mg, KEM은 0.51 mg, 바실러스는 1.05 mg으로대조군보다높게나타났다 (Fig. 8C). 이는적용한미생물에의해사체부패가활발하게일어나 DMS가지속적으로발생됨에따른현상으로사료된다. DMDS: DMDS의경우초기부터고농도로발생하였으며, 시간이경과할수록사체만매몰되어있는반응기에서 DMDS의발생이지속되는것을확인하였다 (Fig. 7D). DMDS의배출잔량은사체만적용한경우에 4.87 mg으로가장높은값을나타냈고, 다른반응기에서는큰차이없이모두비슷한값을가지는것 Fig. 6. According to decomposed rate of carcass, remained the ammonia level (A) and TMA level (B). KEM showed the good removing efficient for TMA, ammonia also decreased by KEM a little. Fig. 7. Tendency of H 2S emission(a), MM emission(b), DMS emission(c) and DMDS emission(d). Usually sulfur compounds were occurred at initial time with high concentration. ( ) Control, ( ) Only media, ( ) Only body, (X) EM, (Ж) KEM, and ( ) Bacillus
348 Kim et al. 으로 확인되었다(Fig. 8D). 미생물의 군집 분포도 변화 분석 DGGE: 분석 결과의 대조군으로 사용하기 위하여 혼합균주 KEM에 대하여 DGGE 분석을 실시하였다(Fig. 9). 광합성 박테 리아, 질소 화합물 악취 제거 균주, 황 화합물 악취 제거 균주와 Fig. 9. The DGGE results from KEM mixture (M, marker; N, negative; 1, fermented mixture I; 2, fermented mixture II; 3, sulfur odor removing mixture; 4, nitrogen odor removing mixture; and 5, photosynthetic bacteria mixture). (A) (B) (C) Fig. 8. According to decomposed rate of carcass, remained the H2S level (A), MM level (B), DMS level (C), and DMDS level (D). KEM showed odor removing efficiency for H2S and MM. Fig. 10. The DGGE results from reactor at initial time (A), after 214 days (B) and after 314 days (C). According to the period of the time, species diversity was reduced (M, marker; C, Control; D, only media; O, only body; E, EM; K, KEM; B, Bacillus; and N, negative).
유용미생물의매몰지조기안정화효과 349 발효균주의 gel상에서의밴드의위치를확인하여다른시료들의 DGGE 분석에있어대조군으로활용하였다. 균의분포도분석결과황화합물악취제거균주를기준으로발효균주 I은 71.8%, 발효균주 II는 59.0%, 광합성박테리아는 65.4%, 그리고질소화합물악취제거균주와는 72.5% 의유사도를보였다. 가축사체의부패가진행됨에따라각반응기내미생물의분포도의종다양성이감소되는것을확인하였으며 (Fig. 10), 밴드상의빨간사각형은 KEM 균주의위치를나타낸다. 실험초기 DGGE 분석결과, 반응기내에서의미생물의종다양성을확인할수있었다. 또한미생물을적용하지않은반응기와각유용미생물을적용한반응기에서의균분포도는상이하게다른것을확인하였다. TMA와황화수소의발생량이증가되는가축사체부패의초기단계에서는 DGGE 분석후대조군의밴드위치와비교해본 결과, 유용미생물의밴드가뚜렷하게관찰되어미생물분포도내의경쟁에서효율적으로생존하고있음을확인하였다. 주기적으로수분과미생물제제를공급해준이후의 DGGE 결과에서는미생물의분포도가전체적으로다양성이감소되는것을확인할수있었다. 이는추가주입한유용미생물제제가우점화가이루어진것으로사료된다. Pyrosequencing: Pyrosequencing 결과사체부패전에는사체만적용한반응기와바실러스적용반응기를제외한모든반응기의초기샘플에서는 uncultured bacteria (39.2%), Bacillus sp. (18.7%), Enterobacter sp. (40.1%), Acinetobacter sp. (24.6%) 등이대부분을차지하고있었다 (Fig. 11). 또한사체만적용반응기와바실러스적용반응기의대부분은병원성미생물로확인되었다. 사체부패가시작된시점에서는 Acinetobacter sp. (31.4%) 와 Enterobacter sp. (21.1%) 와 Pseudomonas sp. (49.4%) 등이우 (A) (B) (C) (D) (E) (F) Fig. 11. The pyrosequencing results from control (A), only media (B), only body (C), EM (D), KEM (E), and Bacillus sp. (F) at initial time.
350 Kim et al. 점화를이루었다 (Fig. 12). 미생물미적용반응기와사체만적용반응기에서는여전히병원성미생물이관찰되었으나, 유용미생물 KEM을적용한경우병원성미생물은확인되지않았다. 사체부패가완료된시점에서는 Planococcaceae (48.1%), Clostridium sp. (62.8%), uncultured bacteria (39.0%) 등이우점화를이루고있었으며균다양성이많이줄어든것을확인하였고, 사체만적용한반응기를제외하고병원성미생물의분포가줄어든것을확인하였다 (Fig. 13.) 결론본연구에서는매몰지모형반응기에유용미생물을적용하 여악취저감효율에따른유용성평가를목적에두고실험하였다. 반응기내의가축사체분해속도는주기적인수분공급이이루어졌을때증가되었으며유용미생물 EM과 KEM을적용하였을때사체가더빠르게분해되는것으로나타났다. 유용미생물을적용하였을경우암모니아와메틸머캅탄의배출잔량이대조군에비해적은것을통하여악취저감효율을확인하였다. 그러나 DMS의경우대조군반응기의배출잔량보다유용미생물적용반응기의배출잔량이많은것으로확인되어저감효율을확인할수없었다. 연구에사용된일반토양자체에이미많은토양미생물이포함되어있고, 또한가축사체내에도미생물들이존재하므로유용미생물을투입하였을때뚜렷한변화는확인할수없었다. 그 (A) (B) (C) (D) (E) (F) Fig. 12. The pyrosequencing results from control (A), only media (B), only body (C), EM (D), KEM (E), and Bacillus sp. (F) after decomposed carcasses.
유용미생물의매몰지조기안정화효과 351 러나일정부분악취제어와부패속도등에효과를증대시켰다. 또한, 악취종류와적용하는유용미생물의종류에따라차이가있지만, 유용미생물을적용하였을때악취저감효율은높은것으로나타났다. 모든매몰지에유용미생물사용은경제성과효율성을고려해야하며, 조기안정화가필요하나토양환경이좋지않을때유용미생물의투입이필요하다. 이때는매몰지침과유용미생물사용지침을새로이만들어야할것이다. 적요본연구를통해유용미생물을적용하여악취저감효과및조기안정화에미치는영향을알아보고자하였다. 유기물분해능 과악취제어능이뛰어난유용미생물 KEM을개발하고, 이미현장에사용중인 EM과바실러스를각각적용하였다. 랩스케일의매몰지모형반응기를제작하여유용미생물을각각적용한경우와적용하지않은경우 ( 미생물을적용하지않고사체만매몰한경우, 배지만적용한경우, 사체만적용한경우 ) 로나누어실험을진행하였다. 주기적인수분공급을해줌으로써가축사체의분해속도가빨라진것을확인하였고, 유용미생물을적용한반응기의사체분해속도가대조군보다빠른것을확인하였다. 가스는총 8개의성분 ( 암모니아, TMA, 황화수소, 메틸머캅탄, DMS, DMDS, CO 2, CH 4) 을중점적으로분석하였으며, 그결과유용미생물을적용하였을경우암모니아와메틸머캅탄에대하여악취저감효율을보였다. (A) (B) (C) (D) (E) (F) Fig. 13. The pyrosequencing results from control (A), only media (B), only body (C), EM (D), KEM (E), and Bacillus sp. (F) after finished the decomposed carcasses.
352 Kim et al. 연구에사용되었던토양내에많은토양미생물과가축사체에도많은미생물들이존재하고있어, 유용미생물을투입하였을때의뚜렷한차이는볼수없었다. 그러나일정부분에한하여악취저감효과와부패속도를증가시키는효과를확인할수있었다. 감사의말 이논문은국립환경과학원 (20120527134) 과미래창조과학부 / 연구재단 (2010-0007473) 의연구비로수행되었습니다. 참고문헌 Amann, R.I., Ludwig, W., and Schleifer, K.H. 1995. Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol. Rev. 59, 143 169. Angelidaki, I. and Ahring, B.K. 1992. Effects of free long-chain fatty acids on thermophilic anaerobic digestion. Appl. Microbiol. Biotechnol. 37, 808 812. Banks, C.J. and Wang, Z. 1999. Development of a two phase anaerobic digester for the treatment of mixed abattoir wastes. Water Sci. Technol. 40, 67 76. Cassidy, D.P. and Hudak, A.J. 2001. Microorganism selection and biosurfactant production in a continuously and periodically operated bioslurry reactor. J. Hazard Mat. 84, 253 264. Castaldi, F.J. and Ford, D.L. 1992. Slurry bioremediation of petrochemical waste sludges. Water Sci. Technol. 25, 207 212. Chun, J., Kim, K.Y., Lee, J.-H., and Choi, Y. 2010. The analysis of oral microbial communities of wild-type and toll-like receptor 2-deficient mice using a 454 GS FLX Titanium pyrosequencer. BMC Microbiol. 10, 101. Eweis, J.B., Ergas, S.J., Chang, D.P.Y., and Schroeder, E.D. 1998. Bioremediation Principles pp. 207 236, McGraw-Hill, USA. Fava, F., Berselli, S., Conte, P., Piccolo, A., and Marchetti, L. 2004. Effects of humic substances and soya lecithin on the aerobic bioremediation of a soil historically contaminated by polycyclic aromatic hydrocarbons (PAHs). Biotechnol. Bioeng. 88, 214 223. Fuller, M.E. and Manning, J.F. 2004. Microbiological changes during bioremediation of explosives-contaminated soils in laboratory and pilot-scale bioslurry reactors. Bioresour. Technol. 91, 123 133. Hanyang University. 2002. Simultaneous Odors and Nutrients Removal Process Development by Soil Origin Microorganisms. Report for National Institute of Environmental Research. Higgins, M.J., Murthy, S.N., Striebig, B., Hepner, S., Yamani, S., Yarosz, D.P., and Toffey, W. 2002. Factors affecting odor production in Philadelphia Water Department Biosolids. Proceedings of Water Environment Federation Odors and Toxic Air Emissions. Ireri, V.R.G., Fabio, F., and Hector, M.P.V. 2008. A review on slurry bioreactors for bioremediation of soils and sediments. Microb. Cell Fact. 7, 5. Janikowski, T.B., Velicogna, D., Punt, M., and Daugulis, A.J. 2002. Use of a two-phase partitioning bioreactor for degrading polycyclic aromatic hydrocarbons by a Sphingomonas sp. Appl. Microbiol. Biotechnol. 59, 368 376. McBean, E.A. and Ro Farquhar, G.J. 1995. Solid waste landfill engineering and design. Prentice Hall, New Jersey, USA. Ministry of environment. 2011. 정부, 전국매몰지일제조사후보완 정비착수. http://www.me.go.kr/web/286/me/common/board/detail. do?boardid=notice_02&decorator=me&idx=176629 Oberbremer, A., Muller-Hurtig, R., and Wagner, F. 1990. Effect of the addition of microbial surfactants on hydrocarbon degradation in a soil population in a stirred reactor. Appl. Microbiol. Biotechnol. 32, 485 489. Pinelli, D., Fava, F., Nocentini, M., and Pasquali, G. 1997. Bioremediation of a polycyclic aromatic hydrocarbon-contaminated soil by using different aerobic batch bioreactor system. J. Soil Contam. 6, 243 256. Salminen, E. and Rintala, J. 2001. Anaerobic digestion of organics solid poultry slaughterhouse waste a review. Bioresour. Technol. 83, 13 26. Wang, S.Y. and Vipulanandar, C. 2001. Biodegradation of naphthalene contaminated soils in slurry bioreactors. J. Environ. Eng. 127, 748 754. Zeev, R. and Aharon, A. 2000. Anaerobic-aerobic process for microbial degradation of retrabromobisphenol. Appl. Environ. Microbiol. 66, 2372 2377. Zhang, C., Hughes, J.B., Nishino, S.F., and Spain, J.C. 2000. Slurry-phase biological treatment of 2,4-dinitrotoluene and 2,6-dinitrotoluene: role of bioaugmentation and effects of high dinitrotoluene concentrations. Environ. Sci. Technol. 34, 2810 2816.