[논문]어병 세균에 대한 소나무과 잎 정유의 항세균 효과

HAM, Youngseok; YANG, Jiyoon; CHOI, Won-Sil; AHN, Byoung-Jun; PARK, Mi-Jin

doi:10.5658/wood.2020.48.4.527

어병 세균에 대한 소나무과 잎 정유의 항세균 효과
Antibacterial Activity of Essential Oils from Pinaceae Leaves Against Fish Pathogens 원문보기

목재공학 = Journal of the Korean wood science and technology, v.48 no.4, 2020년, pp.527 - 547

HAM, Youngseok (Division of Wood Chemistry, Department of Forest Products, National Institute of Forest Science) , YANG, Jiyoon (Division of Wood Chemistry, Department of Forest Products, National Institute of Forest Science) , CHOI, Won-Sil (National Instrumentation Center for Environmental Management, Seoul National University) , AHN, Byoung-Jun (Division of Wood Chemistry, Department of Forest Products, National Institute of Forest Science) , PARK, Mi-Jin (Division of Wood Chemistry, Department of Forest Products, National Institute of Forest Science)

초록
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어병 세균은 어류 양식업의 경제적 피해 뿐만 아니라, 인수공통감염원으로 알려진 전염성 병원균이다. 어병 세균을 제어하기 위해 지속적인 항생제의 사용은 부작용이 수반되기 때문에, 천연 유래 소재의 개발이 요구된다. 본 연구에서는 항생제대체제 개발을 위해 항세균효과가 우수한 침엽수 정유를 발굴하고자 하였다. 소나무과에 속하는 전나무 (Abies holophylla), 곰솔 (Pinus thunbergii), 섬잣나무 (Pinus parviflora), 솔송 (Tsuga sieboldii), 리기테다소나무 (Pinus rigitaeda)의 잎에서 hydro-distillation법을 이용하여 정유를 추출하였으며, 추출된 정유는 어병 세균인 Edwardsiella tarda, Photobacterium damselae, Streptococcus parauberis, Lactococcus garivieae에 대하여 항균력을 평가하였다. 그 결과, 전나무와 곰솔 잎 정유가 그람 음성 세균인 E. tarda와 P. damselae에 대하여 선택적으로 강한 항균력을 나타냈다. GC-MS 분석 결과, 전나무 잎 정유의 주요 성분은 (-)-bornyl acetate (29.45%), D-limonene (20.47%), camphene (11.73%)이고, 곰솔 잎 정유의 주요 성분은 α-pinene (59.81%)으로 각각 확인되었다. 또한, 미량으로 존재하지만 정유와 동일한 효능을 나타내는 유효 성분으로 oxygenated monoterpenes인 neryl acetate, (-)-borneol, (-)-carveol의 세가지 화합물을 구명하였다. 따라서 그람 음성어병세균의 생장억제효과가 우수한 전나무와 곰솔 잎 정유는 사료 첨가제, 수산용 의약품 등 생물학적 제제로 활용 될 수 있을 것으로 사료된다.

Abstract ▼ AI-Helper

Fish pathogens cause not only economic damages to fish farming but also infectious pathogens known as a zoonotic agent. Since the continued use of antibiotics to control fish pathogens entails side effects, materials of natural origin need to be developed. The purpose of this study is to discover coniferous essential oils with excellent antibacterial effects in order to develop antibiotic alternatives. We have extracted essential oils using hydro-distillation from the leaves of Abies holophylla, Pinus thunbergii, Pinus parviflora, Tsuga sieboldii, and Pinus rigitaeda, which are all Pinaceae family. And, we have evaluated antibacterial activity with the extracted essential oils against Edwardsiella tarda, Photobacterium damselae, Streptococcus parauberis, and Lactococcus garvieae, which are fish pathogens. As a result, the essential oils from A. holophylla and P. thunbergii showed the selectively strong antibacterial activity against E. tarda and P. damselae, which are gram-negative bacteria. From GC-MS analysis, it was identified that main component of A. holophylla essential oils are (-)-bornyl acetate (29.45%), D-limonene (20.47%), and camphene (11.73%), and that of P. thunbergii essential oils is α-pinene (59.81%). In addition, we found three compounds: neryl acetate, (-)-borneol, and (-)-carveol, which are oxygenated monoterpenes. These exist in a very small amount but exhibit the same efficacy as essential oil. Therefore, we expect that A. holophylla and P. thunbergii essential oils having excellent growth inhibitory effect against gram-negative fish pathogens can be used as biological products such as feed additives and fishery products.

주제어

표/그림 (5)

표 Table 1. Antibacterial activity of essential oils by the paper disc assay
그림 Fig. 1. Effect of various concentration two essential oils on growth of gram-negative bacteria (A and B) and gram-positive bacteria (C and D).
표 Table 2. Comparative chemical composition of essential oil from five Pinaceae leaves (%)
표 Table 2. (Continued)
그림 Fig. 2. Effect of three oxygenated monoterpenes on growth of gram-negative bacteria (A and B) and gram-positive bacteria (C and D).

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제안 방법

05, and cultured at 28 ˚C for 24 hours. After incubation for 24 hours, the absorbance was measured at 600 nm using an Epoch microplate spectrophotometer (BioTek Instruments, Inc., US) to evaluate the antibacterial activity of essential oils. The control group was treated with 1% DMSO in which the sample was dissolved, and the positive control group was tetracycline (HPLC grade, Product number: 87128, Sigma-Aldrich, Korea), an antibiotic.
For that reason, in this study, we evaluate anti-bacterial activity of essential oils from Pinaceae leaves, which are A. holophylla, P. thunbergii, P. parviflora, T. sieboldii, and P. rigitaeda, against fish pathogens and investigate their own active ingredients. Through this, we tried to investigate whether essential oils from Pinaceae leaves can be used as a prevention of fish pathogens or as a medicine for fisheries.
In this study, we evaluated the activity of the terpene-based single compound constituting the essential oil, and sought to identify an active ingredient exhibiting similar physiological activity to the essential oils. We carried out antibacterial screening of various terpene-based single compounds (0.
72%, and the antibacterial effect was evaluated with the paper disc diffusion method. The presence of antibacterial activity of essential oils was confirmed by measuring the size of the ring formed by exposing 4 types of fish pathogens to a paper disc containing essential oils for 2 days (Table 1). A clear zone arising around the paper disc is formed because bacteria cannot grow in the area, so the size of the growth inhibiting ring indicates an index of antibacterial activity (Djabou et al.
The purpose of this study was to evaluate the possibility of substituting essential oil, a natural product, for antibiotics used to treat infectious fish diseases caused by bacteria to compensate for side effects such as the emergence of resistant strains, which have been pointed out as a disadvantage of using antibiotics. We evaluated antibacterial activity of the five essential oils extracted from the leaves of pine family, and noticed that A.

대상 데이터

Edwardsiella tarda FP5060, Photobacterium damselae FP4101, Lactococcus garvieae FP5245, and Streptococcus parauberis FP3287 were received from Marine and Fisheries Life Resources, National Institute of Fisheries Sciences, and they were stored in 25% glycerol stock at -40˚C. BHI agar was prepared by adding 15 g/L agar to BHI broth (Bacto^TM brain heart infusion, Product number: 237500, BD Biosciences Korea Ltd.

데이터처리

, Chicago, Illinois, USA). Significance levels were set at 95% confidence intervals for Tukey test or paired-t-test.
Standard deviation was calculated from four independent experiments. Statistical analysis was performed using a Tukey test. Values that differ from the control with the 95% confidence level are marked with a star (A.
Standard deviation was calculated from four independent experiments. Statistical analysis was performed using a paired-t-test. Values that differ from the control with the 95% confidence level are marked with a star on the top of bars.

성능/효과

24%). A total of 17 compounds were found in both essential oils of A. holophylla and P. thunbergii leaves, and 12 were monoterpene and 5 were sesquiterpene. From these results, we identified that (-)-bornyl acetate and α-pinene account for the largest proportion of the two essential oils extracted by hydro-distillation in this study.
As a result of the paper disc assay, E. tarda and P. damselae, the gram-negative bacteria, showed high sensitivity to essential oils of A. holophylla and P. thunbergii leaves. In contrast, the gram-positive bacteria L.
parauberis. From these results, we noticed that the essential oils of A. holophylla and P. thunbergii leaves had a strong antibacterial effect against fish pathogens, and were particularly effective against E. tarda and P. damselae.
, 2016). From these studies, it also showed a synergistic effect of antibacterial activity between neryl acetate, (-)-carveol, (-)-borneol, and other components in the essential oils of A. holophylla and P. thunbergii leaves, and it is presumed to have high antibacterial activity when compared to P. rigitaeda.
However, studies on fish pathogens with these compounds have not been conducted, and such previous studies have not provided results on specific antibacterial activity by species classification. Given these results, this study suggests the possibility of selectively controlling gram-negative fish pathogens by using neryl acetate, (-)-borneol and (-)-carveol, which are classified as oxygenated monoterpene.
In addition, 0.035% concentration of A. holophylla leaves essential oil showed inhibitory effect on the growth of about 93% and P. thunbergii leaves essential oil did about 81%.
Interestingly, however, the essential oils from A. holophylla, P. thunbergii, P. rigitaeda, and T. sieboldii leaves tested in this study were more effective and showed high antibacterial activity against gram-negative bacteria (E. tarda and P. damselae) than gram-positive bacteria (L. garvieae and S. parauberis) (Table 1). Particularly, MIC concentrations of E.
P. thunbergii leaves essential oil had the highest ratio of α-pinene at 59.81%, followed by caryophyllene oxide (5.44%), D-limonene (4.34%), and β-myrcene (4.24%).
parviflora (8 mm) in order. The antibacterial effect of tested essential oils against L. garvieae was lower than that of E. tarda and P. damselae, and among 5 essential oils, A. holophylla essential oil was 12.75 mm, which showed higher antibacterial activity than other essential oils. For S.
thunbergii leaves, which have a strong antibacterial activity against gram-negative bacteria, were analyzed as terpene-based compounds, including monoterpene and sesquiterpene. The main components of A. holophylla leaves essential oil were 22 monoterpene (79.82%) and 7 sesquiterpene (7.21%), and the main constituents of P. thunbergii leaves essential oil were 26 monoterpene (80.94%) and 9 sesquiterpene (14.00%). The main components of A.
The yields of each essential oil extracted from A. holophylla, P. thunbergii, P. parviflora, T. sieboldii, and P. rigitaeda leaves were found to be 2.99%, 0.93%, 0.08%, 0.15%, 0.72%, and the antibacterial effect was evaluated with the paper disc diffusion method.
The purpose of this study was to evaluate the possibility of substituting essential oil, a natural product, for antibiotics used to treat infectious fish diseases caused by bacteria to compensate for side effects such as the emergence of resistant strains, which have been pointed out as a disadvantage of using antibiotics. We evaluated antibacterial activity of the five essential oils extracted from the leaves of pine family, and noticed that A. holophylla and P. thunbergii leaves essential oils showed strong growth inhibitory effects against gram-negative fish pathogens, E. tarda and P. damselae. As a result of analyzing the components of these two essential oils by GC-MS, they mainly consisted of monoterpene-based compounds, and the main components were identified as (-)-bornyl acetate (29.

후속연구

In the case of gram-negative bacteria with high resistance to hydrophobic molecules, it has been reported that some hydrophobic compounds such as essential oils can slowly pass through the porin protein present in the cell wall (Plesiat and Nikaido, 1992; Bock and Sawers, 1996). Based on these previous studies, further studies are needed about additional mechanisms for the gram-negative fish pathogens of essential oil and three oxygenated terpenes obtained in this study. In this study, we found a material that is selectively applied only to a specific species among various fish pathogens that are problems in aquaculture.

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표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

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