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연구개발결과
1세부과제에서는 12종 사료원료의 메탄발생기여도 차이를 in vitro batch culture 방법으로 조사하였다. 60ml serum bottle에 30ml 배양액과 200mg의 원료를 넣어 24시간 조사한 결과, 사료원료들 간의 메탄발생량은 3개의 그룹으로 구분되었다. 고매탄군은 corn powder과 corn germ meal과 같은 고에너지 사료원료들이었고, 중메탄군은 고단백질 사료원료로서 에너지 부족에 의한 단백질 발효효율이 낮아졌기 때문으로 보인다. 저메탄군은 timothy, whole crop ric

연구개발결과
1세부과제에서는 12종 사료원료의 메탄발생기여도 차이를 in vitro batch culture 방법으로 조사하였다. 60ml serum bottle에 30ml 배양액과 200mg의 원료를 넣어 24시간 조사한 결과, 사료원료들 간의 메탄발생량은 3개의 그룹으로 구분되었다. 고매탄군은 corn powder과 corn germ meal과 같은 고에너지 사료원료들이었고, 중메탄군은 고단백질 사료원료로서 에너지 부족에 의한 단백질 발효효율이 낮아졌기 때문으로 보인다. 저메탄군은 timothy, whole crop rice straw, rice straw와 같은 조사료원이었고, in vivo 실험결과에서 메탄발생량이 가장 많은 것과는 상반되는 보고로서, 결과 해석에 주의가 요구된다. 향후 저메탄사료의 배합에 활용하기 위해서는 단백질은 물론 상대적으로 에너지 함량도 높은 단백질추출 부산물들의 메탄발생기여도를 in vitro batch culture 방법을 이용해 추가 비교한 결과, soybean meal과 lupin seed가 가장 높았고, rapeseed meal과 corn gluten feed가 다음, luten, cotton seed meal가 가장 낮았다. 다음 실험으로 6개의 다른 수입 모선에서 채취한 총 40종의 사료원료를 총 7회에 걸쳐 in vitro batch culture 방법으로 평가한 결과, 밀>옥수수>congluten feed>soybean meal>rapeseed meal>palm kernel meal> pineapple meal의 순으로 메탄발생량이 낮아졌다. 다음 in vitro batch culture 실험에서는 주요 에너지사료인 옥수수, 보리와 밀의 메탄발생기여도를 비교한 결과, 보리가 옥수수와 밀보다 메탄발생량이 높았고 in vivo 실험에서도 보리, 밀, 옥수수가 각각 42.7, 40.0,38.8kg/head/year 이었다. 이상의 시험결과들을 기초로 배합된 저메탄 예측사료와 고메탄 예측사료를 거세한우에게 급여한 결과, 25%의 메탄이 저감하는 것을 확인하였다. 본 과제는 국내에서 처음으로 수행된 저메탄사료 개발연구로서, 향후 축산 현장에 미칠 영향력을 생각할 때, 추가적인 실험들이 반드시 필요한 분야로 생각된다.
제2세부과제는 축산물품질평가원의 등급 통계자료를 분석하여 2008∼2010년 3년간의 육질 등급의 분포와 거세우 비육농가에서 문제가 되는 육량 C등급의 비율을 비교하여 어느 시기에 고온의 영향을 받는지 추정하여 본 결과, 해발고도가 300~800m로 여름철 고온기가 상대적으로 짧은 중산간지대인 평창지역의 거세우 1등급이상 출현율은 3년간 평균 86.9%로 전국 평균 약 78%에 비해 높은 편이었으며, 10월부터 11월 사이에 4~5% 가량 하락하여 고온기 피해가 전국 데이터와 다소 다른 것으로 추정되었다. 다음으로 실제 현장에서 고운기에 스트레스가 심할 것으로 추정되는 경북 영주시와 고온스트레스가 없는 강원도 평창군에서의 사양실험을 동시에 수행하였다. 농후사료 급여량은 2004년 국립축산과학원에서 제시한 고급육생산 사양프로그램을 적용하여 육성기인 11개월령까지 농후사료 체중비 1.6%, 비육전기인 15개월령까지 체중의 1.7%, 이후 16개월령 비육중기부터 28개월령 비육후기까지 1.8% 내외로 농협사료에서 생산된 시판 배합사료를 급여하였고, 육성기는 이탈리안 라이그라스, 티모시 건초, 비육전기 이후 볏짚을 자유채식 시킨 결과, 출하시 체중은 영주와 대관령지역 각각 700.3 및 708.7㎏이었고 등지방 두께는 영주지역게서 사육된 시험축이 2㎜정도 두꺼워 육량 C 등급이 약간 높게 나왔으나 큰 차이는 아니었고, 육색이나 지방색은 대관령 지역에서 사육된 시험축이 다소 엷은 경향이었다. 한편 근내지방은 두 지역간에 차이가 거의 없었고 육질 1등급 이상 출현 비율은 대관령지역에서 사육된 시험축에서 1두가 나왔으나 1++ 등급이 영주는 없고 대관령에서 1두가 있어 전반적인 경제성에서는 큰 차이가 없는 것으로 분석되었다.
제1협동과제는 고온스트레스 저감제제가 반추위 발효에 미치는 영향 및 저감제제의 적정 혼합 비율을 구명하기 위해 실시된 실험으로 첫째, 고온스트레스 저감제제가 반추위 발효에 미치는 영향 및 저감제제의 적정 혼합 비율을 in vitro 실험으로 구명한 결과, 보호지방과 콜린의 혼합첨가가 반추위 내 pH 하강을 완충시키고 반추위 발효 가스발생량을 조절할 수 있는 것으로 판단되었다. 한우 고온스트레스 저감형 최적 사료 영양조건 구명을 위한 두 번째 in vitro 발효 실험에서는 보호지방 0.05%, 저감제제 0.05% 수준의 급여가 반추위 발효에 대한저해현상 없이 발효열을 감소시킬 것으로 판단되었고, 이러한 결과를 바탕으로 in vivo 대사시험을 실시한 결과에서도 체온 변화에 영향을 주는 것을 확인하였고, 간접적으로는 반추위 발효 성상에도 긍정적인 효과가 있었다. 최종실험에서는 육성기, 비육기(비육전기, 비육후기)의 거세우를 공시하여 고온스트레스 저감제제의 하절기 현장 기호성 테스트를 실시하였다. 육성기 및 비육기의 growth performance (사료섭취량, 일당증체량 및 사료요구율)는 고온스트레스 저감제제 급여로 통계적 유의성은 없었으나 개선되는 경향을 보였으며(p>0.05), 특히 비육후기에는 유의성 있게 저감제제 급여구가 사료섭취량 부분에서 뚜렷한 효과를 나타내었다. 위 결과를 바탕으로 고온스트레스 저감제제가 기호성을 개선하여 사료섭취량을 개선 할 수 있으며, 이에 따라 증체율에도 간접적으로 기여할 수 있는 가능성을 확인하였다.
제2협동과제에서는 지구온난화 대비 고온기 한우 영양소 요구량 설정 연구를 수행하기 위한 환경 제어 장치 구축를 구축하였고, 현재 한국사양표준 내 유지를 위한 한우의 TDN요구량 재설정응 위한 실험을 수행한 결과, 유지를 위한 한우 번식우의 TDN 요구량의 100%를 급여한 대조구에서 일당증체량이 -31.25g이 나타났고 (체중 감소), 유지를 위한 한우 육성우의 TDN 요구량의 100%를 급여한 대조구에서 일당증체량이 -46.4g이 나타났다(체중 감소). 또한 유지를 위한 한우 비육우의 TDN 요구량 실험에서도 100%를 급여한 대조구에서 일당증체량이 –69.6g이 나타났다(체중 감소). 따라서 지구온난화(고온기) 시 한우 번식우 유지를 위한 TDN 요구량은 현재 한국사양표준 TDN 요구량 대비 11.24%를 증량, 지구온난화(고온기) 시한우 육성우 유지를 위한 TDN 요구량은 현재 한국사양표준 TDN 요구량 대비15.0%를 증량, 지구온난화(고온기) 시 한우 비육우 유지를 위한 TDN 요구량은 현재 한국사양표준 TDN 요구량 대비 11.5%를 증량하여 급여하는 방향으로 한국사양 표준의 영양소요구량을 재 설정하여야 한다고 판단된다.
제3협동과제는 장내발효 메탄저감을 위한 메탄지표 및 사료첨가제개발을 위해 첫 번째로 현장에서 가장 많이 사용되는 15종 사료에 대한 메탄발생기여도를 in vitro batch culture 방법으로 조사한 결과, 발효될 수 있는 기질이 풍부한 곡류에서 methane 발생량이 높았다. 박류와 같이 단백질 함량이 높은 원료사료의 경우 반추위 미생물에 의한 분해, 생성된 NH₄⁺가 Methane의 기질인 이산화탄소와 결합함으로써 methane 생성량이 감소하였다. 볏짚의 세포벽을 구성하는 성분은 반추위 미생물에 의해 쉽게 분해되기 어려운 lignin 성분을 많이 포함하고 있어서 이용성이 낮아 methane 발생량이 낮았다. 다음 실험으로서 주요 경제사료 5종의 in vitro batch culture 실험을 수행한 결과, 단백피, 옥배아박, 타피오카전분박처럼 원료 내 전분함량이 높은 사료들이 팜박, 채종박같이 전분함량이 거의 없는 원료들보다 유의적으로 메탄발생량이 높았다. 본 실험의 특징은 in vitro continuous culture system을 이용하여 주요 축우 원료 9종의 메탄발생량을 측정하고, 이를 NIRs system에 적용하여 원료사료별 메탄발생 예측이 가능하도록 메탄발생예측 지수 도출시도한 것이다. 그 결과, 소맥 > 옥수수 > 대두박 > 알팔파 > 볏짚 > 채종박 > 야자박 > 팜박 > 단백피 순의 평균값을 보였고, NIR system을 이용한 예측값과 비교하였을 때, R square가 0.76로 정량가능 수준이기는 하나, 메탄발생지수 예측 오차범위가 넓어 보다 정확한 예측 모델을 도출하기 위해서는 각 원료당 모선 n 수를 100~1000 개 이상의 데이터를 축적해야 할 필요성이 있다. 최종적으로 Laser methane detector를 이용하여 착유우 농가의 메탄모니터링 정확성을 검증한 결과, 메탄저감 첨가제나 그린사료의 효과를 검증하는 방법으로 사용하기에 적절하다고 사료된다

Abstract ▼

An experiment was conducted to evaluate the potential producing methane for 12 feed materials in batch cultures of mixed rumen microorganisms. Serum bottles containing 200mg of the feed material tested and 30ml of buffered rumen fluid (24ml

An experiment was conducted to evaluate the potential producing methane for 12 feed materials in batch cultures of mixed rumen microorganisms. Serum bottles containing 200mg of the feed material tested and 30ml of buffered rumen fluid (24ml rumen fluid + 6ml culture medium) were incubated at 39℃ for 24hr. Tested feeds have different CH4 production. The major methane production arised from energy-rich feedstuffs, such as corn powder and corn germ meal. The moderate methanogenesis was observed in protein sources caused by the low energetic efficiency of protein metabolization. In contrast to the results of in vivo experiment, forage such as the timothy, whole crop rice straw and rice straw showed low methane production.
The next experiment aimed to identify the in vitro fermentation characteristics of protein feed sources. The major methane production arised from soybean meal and lupin seed. The moderate methanogenesis was observed from rapeseed meal and corn gluten feed. on the contrary luten, cotton seed meal showed the low methane production. The third experiment was conducted to evaluate the potential producing methane for 40 feed materials in batch cultures of mixed rumen microorganisms. Among the groups, wheat and corn were highest in methane production, followed by congluten feed, soybean meal, rapeseed meal, palm kernel meal, pineapple meal.
The fourth study was conducted to investigate the effects of grain sources on the characteristics of ruminal fermentation, gas production and microbial population. In the first in vitro experimet, ruminal fluid was collected from two cannulated Hanwoo steers(average BW 374kg) before their morning feeding on the day. Treatments were Corn, Barley, Wheat and Mix(Corn : Barley : Wheat = 1 : 1 : 1). Total gas production was lowest(P < 0.05) for Corn than other treatments at 6 and 12h. Methane production was higher(P < 0.05) for Barley than corn and wheat at 12h. The concentration of total VFA was lowest(P < 0.05) for Corn than other treatments at 6 and 12h. The acetate : propionate ratio was higher(P < 0.05) for barley than other treatments at 12h.
The fifth in vivo experiment focused on comparison in digestibility, rumen fermentation, methane production and microbial population from growing Hanwoo. Twelve Hanwoo steers(BW = 355±5.35kg) were used in a 3 × 3 Latin square design with four treatments. Treatments were corn(60 %), barley(60 %), wheat(60 %) and mix(corn : barley : wheat = 20 : 20 : 20%). Nutrient intake were higher(P < 0.05) for Wheat and Mixed than Corn and Barley. Nutrient digestibility were lowest(P < 0.05) for Corn than other treatment on CP. However grain sources did not affect digestibilities of DM, Crude fiber and Crude ash. Ruminal pH was higher(P < 0.05) for Corn and Barley than Wheat and Mixed. NH3-N was higher(P< 0.05) for Wheat than other treatments. Total VFA was lowest(P < 0.05) for Corn than other treatments and acetate : propionate ratio was high for Barley than other treatments. Ruminal microbial population of methanogens was high for Barley than other treatments. Methane emission averaged 38.8, 42.7 and 40.0kg/head/year for corn, Barley and Wheat, respectively. The final in vivo experiment was performed to evaluate low and high methane emission feeds formulated by the results obtained from the previous experiments. Hanwoo fed low methane emission feed showed low methane production up to 25% rather than that for high methane emission feed.
< Development of technology for improving productivity of Hanwoo steer in high temperature>
This study was conducted to investigate differences by simultaneous raising in high or moderate climate region on Hanwoo steers. In experiment 1, effective supplementary materials were screened and selected. Then, subsequent experiment was employed to figure out the optimum mixture condition using Box Behnken Design which is one of fractional factorial designs. Stevioside, cinnamon and organic zinc which were used as a fixed factor and protected fat, choline and yeast used as variables. The optimal mixture condition was simulated based on volatile fatty acid production which is related to rumen fermentative parameter. Mixed additive give better result for decreasing rumen fermentative heat compared to single additive supplementation based on total gas and volatile fatty acid productions in the rumen fermentation that was performed with in vitro technique. In addition, the desirable supplementary level without any negative effect on rumen microbial fermentation was found. Feed intake was particularly improved during late fattening period rather than early and mid fattening period and this might be caused by its palatability. As a result, the present results suggested that the developed additive could relive heat stress effectively and show a possibility to improve growth performance.

This study was conducted to develop a dietary supplement to relieve high temperature stress in Hanwoo using protected fat, probiotics, choline, stevioside, cinnamon and organic zinc. In experiment 1, effective supplementary materials were screened and selected. Then, subsequent experiment was employed to figure out the optimum mixture condition using Box Behnken Design which is one of fractional factorial designs. Stevioside, cinnamon and organic zinc which were used as a fixed factor and protected fat, choline and yeast used as variables. The optimal mixture condition was simulated based on volatile fatty acid production which is related to rumen fermentative parameter. Mixed additive give better result for decreasing rumen fermentative heat compared to single additive supplementation based on total gas and volatile fatty acid productions in the rumen fementation that was performed with in vitro technique. In addition, the desirable supplementary level without any negative effect on rumen microbial fermentat ion was found at 0.1%. And we conducted in vivo experiment to evaluate the effect of additives with different forage : concentration ration (6:4 and 4:6) on body temperature and the rumen fermentation. Four rumen cannulated Hanwoo steers (body wt. 400 kg±30 kg) were used in 4x4 latin square experimental design. The dietary supplementation significantly decreased internal rectum and rumen temperatures (p<0.05), and increased propionate production (p<0.05). In farm experiment, it was found that the dietary supplementation had positive effects on growth performance such as feed intake, body weight and feed efficiency during summer season even though no statical differences were found. Feed intake was particularly improved during late fattening period rather than early and mid fattening period and this might be caused by its palatability. As a result, the present results suggested that the developed additive could relive heat stress effectively and show a possibility to improve growth performance.

Ⅰ. 1st year (2010)
- Establishment of environmental modulation system for high temperature stress related research
A global warming research building including environmental modulation system with 6 metabolism cages for large animal and 2 independent chambers modulating temperature and humidity was established at Daegu University Research Farm.
Ⅱ. 2nd year (2011)
- Establishment of nutrient requirement for Hanwoo cow under high temperature against global warming
This study was carried out to establish nutrient requirements for Korean native cows under high temperature for global warming. Four Hanwoo cows weighing 491±85.6kg were used within 4×4 Latin square design. Hanwoo cows were fed four different energy level diets; 100% (control), 100%, 115% and 130% of TDN levels of Hanwoo cows based on Korea Feeding Standard for Hanwoo using rice straw and commercial concentrate. Hanwoo cows in the control were housed with no stress (temperature 24℃ and humidity 60%; temperature and humidity index (THI) =71.46), whereas Hanwoo cows in the other groups were under 30℃ in temperature and 70% in humidity (THI = 81.38). Apparent digestibility of dry matter was significantly (p<0.05) higher in control (81.64%) and T100 (81.39%) than T115 (78.23%) and T130 (78.15%). High THI and different energy intake levels did not affect rectal temperature of Hanwoo cows. Blood cortisol concentration was not affected by the treatment, except for T130 being numerically higher than the other groups. Average daily gain for T100(-56.00g) was lower (p<0.05) than control (-31.25g), T115(35.50g) and T130(77.25g), respectively, indicating that TDN requirement for maintenance of Hanwoo cows under high temperature condition should be increased to 11.24% based on TDN requirement for Hanwoo cows in the Korean Feeding Standard for Korean native cows.
Ⅲ. 3rd year (2012)
- Establishment of nutrient requirement for growing Hanwoo cattle under high temperature against global warming
Four growing Hanwoo cattle weighing 200±11.7kg were used within 4×4 Latin square design to establish nutrient requirements for growing Hanwoo cattle under high temperature. The growing Hanwoo cattle were fed four different energy level diets; 100% (control), 100%, 115% and 130% of TDN levels of growing Hanwoo based on Korea Feeding Standard for Hanwoo using timoth hay and commercial concentrate. The growing Hanwoo in the control were housed with no stress (temperature 24℃ and humidity 60%; temperature and humidity index (THI) =71.46), whereas the growing Hanwoo in the other groups were under 30℃ in temperature and 70% in humidity (THI = 81.38). True digestibility of dry matter was significantly (p<0.05) higher in control (81.48%) and T100 (79.07%) than T115 (76.96%) and T130 (75.97%). High THI and different energy intake levels did not affect rectal temperature of the growing Hanwoo. Blood cortisol concentration was not affected by the treatment, except for T115 being numerically higher than the other groups. Average daily gain for T100 (-68.8g) was lower than control (-46.4g, numerically), T115(20.5g, p<0.05) and T130(47.9g, p<0.05), respectively, indicating that TDN requirement for maintenance of growing Hanwoo cattle under high temperature condition should be increased to 15.0% based on TDN requirement for growing Hanwoo cattle in the Korean Feeding Standard for Hanwoo.
Ⅳ. 4th year (2013)
- Establishment of nutrient requirement for fattening Hanwoo cattle under high temperature against global warming
Four fattening Hanwoo cattle weighing 247±13.5kg were used within 4×4 Latin square design to establish nutrient requirements for fattening Hanwoo cattle under high temperature. The fattening Hanwoo cattle were fed four different energy level diets; 100% (control), 100%, 115% and 130% of TDN levels of fattening Hanwoo based on Korea Feeding Standard for Hanwoo using rice straw and commercial concentrate. The fattening Hanwoo in the control were housed with no stress (temperature 24℃ and humidity 60%; temperature and humidity index (THI) = 71.46), whereas the fattening Hanwoo in the other groups were under 30℃ in temperature and 70% in humidity (THI = 81.38). True digestibility of dry matter was not significantly (p>0.05) differ between control and the treated groups (69.04 ∼72.30%). High THI and different energy intake levels did not affect rectal temperature (38.23∼38.32℃) of the fattening Hanwoo. Blood IgG and thyroxine concentrations were not affected by the treatments. Average daily gain for T100 (-69.6g) was lower than control (-44.6g, numerically), T115(44.6g, p<0.05) and T130(83.3g, p<0.05), respectively, indicating that TDN requirement for maintenance of fattening Hanwoo cattle under high temperature condition should be increased to 11.5% based on TDN requirement for fattening Hanwoo cattle in the Korean Feeding Standard for Hanwoo.

An experiment was conducted to evaluate the potential producing methane for 12 feed materials in batch cultures of mixed rumen microorganisms. Serum bottles containing 200mg of the feed material tested and 30ml of buffered rumen fluid (24ml rumen fluid + 6ml culture medium) were incubated at 39℃ for 24hr. Tested feeds have different CH4 production. The major methane production arised from energy-rich feedstuffs, such as corn powder and corn germ meal. The moderate methanogenesis was observed in protein sources caused by the low energetic efficiency of protein metabolization. In contrast to the results of in vivo experiment, forage such as the timothy, whole crop rice straw and rice straw showed low methane production.
The next experiment aimed to identify the in vitro fermentation characteristics of protein feed sources. The major methane production arised from soybean meal and lupin seed. The moderate methanogenesis was observed from rapeseed meal and corn gluten feed. on the contrary luten, cotton seed meal showed the low methane production. The third experiment was conducted to evaluate the potential producing methane for 40 feed materials in batch cultures of mixed rumen microorganisms. Among the groups, wheat and corn were highest in methane production, followed by congluten feed, soybean meal, rapeseed meal, palm kernel meal, pineapple meal.
The fourth study was conducted to investigate the effects of grain sources on the characteristics of ruminal fermentation, gas production and microbial population. In the first in vitro experimet, ruminal fluid was collected from two cannulated Hanwoo steers(average BW 374kg) before their morning feeding on the day. Treatments were Corn, Barley, Wheat and Mix(Corn : Barley : Wheat = 1 : 1 :1). Total gas production was lowest(P < 0.05) for Corn than other treatments at 6 and 12h. Methane production was higher(P < 0.05) for Barley than corn and wheat at 12h. The concentration of total VFA was lowest(P < 0.05) for Corn than other treatments at 6 and 12h. The acetate : propionate ratio was higher(P < 0.05) for barley than other treatments at 12h.
The fifth in vivo experiment focused on comparison in digestibility, rumen fermentation, methane production and microbial population from growing Hanwoo. Twelve Hanwoo steers(BW = 355±5.35kg) were used in a 3 × 3 Latin square design with four treatments. Treatments were corn(60 %), barley(60 %), wheat(60 %) and mix(corn : barley : wheat = 20 : 20 : 20%). Nutrient intake were higher(P < 0.05) for Wheat and Mixed than Corn and Barley. Nutrient digestibility were lowest(P < 0.05) for Corn than other treatment on CP. However grain sources did not affect digestibilities of DM, Crude fiber and Crude ash. Ruminal pH was higher(P < 0.05) for Corn and Barley than Wheat and Mixed. NH3-N was higher(P < 0.05) for Wheat than other treatments. Total VFA was lowest(P < 0.05) for Corn than other treatments and acetate : propionate ratio was high for Barley than other treatments. Ruminal microbial population of methanogens was high for Barley than other treatments. Methane emission averaged 38.8, 42.7 and 40.0kg/head/year for Corn, Barley and Wheat, respectively. The final in vivo experiment was performed to evaluate low and high methane emission feeds formulated by the results obtained from the previous experiments. Hanwoo fed low methane emission feed showed low methane production up to 25% rather than that for high methane emission feed.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 요약문 ... 6
• SUMMARY ... 11
• 목차 ... 18
• 제 1 장 서 론 ... 19
• 제 2 장 국내외 기술개발 현황 ... 20
• 제 3 장 연구개발수행 내용 및 결과 ... 23
• 제1절 가축의 장내발효 메탄 저감사료 개발 ... 23
• 제2절 고온기 한우거세우의 생산성 향상기술 개발 ... 63
• 제3절 고온기 한우대사생리 변화 구명 및 고온스트레스 저감형사료 이용기술 개발 ... 69
• 제4절 지구 온난화 대비 고온기 한우 영양소 요구량 설정 연구 ... 106
• 제 4 장 연구개발목표 달성도 및 대외기여도 ... 166
• 1절. 목표대비 달성도 ... 166
• 2절. 정량적 성과 ... 168
• 제 5 장 연구개발결과의 활용계획 ... 169
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 170
• 제 7 장 기타 중요 변동사항 ... 170
• 제 8 장 국가과학기술종합정보시스템에 등록한 연구장비 현황 ... 170
• 제 9 장 참고문헌 ... 171
• 끝페이지 ... 181