JP2769168B2 - Medium for detecting beer harmful bacteria - Google Patents
Medium for detecting beer harmful bacteriaInfo
- Publication number
- JP2769168B2 JP2769168B2 JP63296097A JP29609788A JP2769168B2 JP 2769168 B2 JP2769168 B2 JP 2769168B2 JP 63296097 A JP63296097 A JP 63296097A JP 29609788 A JP29609788 A JP 29609788A JP 2769168 B2 JP2769168 B2 JP 2769168B2
- Authority
- JP
- Japan
- Prior art keywords
- bacteria
- medium
- beer
- oleic acid
- harmful bacteria
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 241000894006 Bacteria Species 0.000 title claims description 49
- 235000013405 beer Nutrition 0.000 title claims description 30
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 claims description 26
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 claims description 26
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 claims description 26
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 claims description 26
- 239000005642 Oleic acid Substances 0.000 claims description 26
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 claims description 26
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 claims description 26
- 239000002609 medium Substances 0.000 description 20
- 230000000694 effects Effects 0.000 description 14
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 9
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 9
- 238000012360 testing method Methods 0.000 description 7
- 241000193830 Bacillus <bacterium> Species 0.000 description 4
- 241000589516 Pseudomonas Species 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 240000001929 Lactobacillus brevis Species 0.000 description 3
- 235000013957 Lactobacillus brevis Nutrition 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 241000588914 Enterobacter Species 0.000 description 2
- 241000589565 Flavobacterium Species 0.000 description 2
- 241000588731 Hafnia Species 0.000 description 2
- 241000588748 Klebsiella Species 0.000 description 2
- 241000588915 Klebsiella aerogenes Species 0.000 description 2
- 241000192001 Pediococcus Species 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- CJNBYAVZURUTKZ-UHFFFAOYSA-N hafnium(IV) oxide Inorganic materials O=[Hf]=O CJNBYAVZURUTKZ-UHFFFAOYSA-N 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- 241000589291 Acinetobacter Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000186216 Corynebacterium Species 0.000 description 1
- 244000078127 Eleusine coracana Species 0.000 description 1
- 235000013499 Eleusine coracana subsp coracana Nutrition 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 241000589538 Pseudomonas fragi Species 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 235000002079 ragi Nutrition 0.000 description 1
- 238000011160 research Methods 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Distillation Of Fermentation Liquor, Processing Of Alcohols, Vinegar And Beer (AREA)
Description
【発明の詳細な説明】 ［産業上の利用分野］ 本発明はビール有害菌の検出用培地に関する。Description: TECHNICAL FIELD The present invention relates to a medium for detecting harmful bacteria of beer.
［従来の技術、発明が解決しようとする課題］ ビール有害菌としてグラム陽性菌のほかグラム陰性菌
も知られている。[Prior Art and Problems to be Solved by the Invention] Gram-positive bacteria as well as gram-negative bacteria are known as beer harmful bacteria.
ビール製造過程において見出されるビール有害菌は通
常、細胞活性が低下しているため、その増殖が遅く、検
出するまでに長時間を要する。Beer harmful bacteria found in the process of beer production usually have a low cell activity, so their growth is slow and it takes a long time to detect them.
ビール有害菌を特異的に、かつ迅速に検出する手段に
ついて従来は十分な検討がなされていなかった。そのた
め、これらビール有害菌を効果的に検出するための培地
の開発が望まれていた。A means for specifically and rapidly detecting harmful beer bacteria has not been sufficiently studied. Therefore, development of a medium for effectively detecting these beer harmful bacteria has been desired.
［課題を解決するための手段］ 本発明者らは、上記課題を解決すべく検討を重ねた結
果、特定の高級脂肪酸を添加した培地を用いると、ビー
ル有害菌の増殖が促進されることを見出し、かかる知見
に基いて本発明に到達した。[Means for Solving the Problems] As a result of repeated studies to solve the above problems, the present inventors have found that the use of a medium containing a specific higher fatty acid promotes the growth of beer harmful bacteria. The present invention has been accomplished based on the findings and the findings.
すなわち、本発明はオレイン酸５〜100ppmを含有せし
めたビール有害菌の検出用培地を提供するものである。That is, the present invention provides a culture medium for detecting harmful beer bacteria containing 5 to 100 ppm of oleic acid.
本発明が適用されるビール有害菌には特に制限はない
が、具体的にはグラム陽性菌であるラクトバチルス（La
ctobacillus）属に属する細菌、例えばラクトバチルス
・ブレビス（Lactobacillus brevis）AHU 1508,コリネ
バクテリウム（Corynebacterium）属に属する細菌，ペ
ディオコッカス（Pediococcus）属に属する細菌、例え
ばペディオコッカス・セレビシア（Pediococcus cerev
isiae），バチルス（Bacillus）属に属する細菌、例え
ばバチルス・ノンスポラ（Bacillus nonspore）等、グ
ラム陰性菌であるエンテロバクター（Enterobacter）属
に属する細菌，ハフニア（Hafnia）属に属する細菌，ク
レブシエラ（Klebsiella）属に属する細菌、例えばクレ
ブシエラ・アエロゲネス（Klebsiella aerogenes）ATC
C 15050,フラボバクテリウム（Flavobacterium）属に属
する細菌，シュードモマス（Pseudomonas）属に属する
細菌、例えばシュードモナス・フラギ（Pseudomonas f
ragi）IFO 3458,アシネトバクター（Acinetobacter）属
に属する細菌等を挙げることができる。No particular limitation is imposed on the harmful beer bacteria to which the present invention is applied, and specifically, Lactobacillus ( La
Bacteria belonging to the genus ctobacillus , such as Lactobacillus brevis AHU 1508, bacteria belonging to the genus Corynebacterium , bacteria belonging to the genus Pediococcus , such as Pediococcus cerevia
Isiae), Bacillus (Bacillus) bacteria belonging to the genus, for example, Bacillus Nonsupora (Bacillus nonspore) or the like, bacteria belonging to the Enterobacter (Enterobacter) genus are Gram-negative bacteria, hafnia (Hafnia) bacterium belonging to the genus Klebsiella (Klebsiella) Bacteria belonging to the genus, for example, Klebsiella aerogenes ATC
C 15050, Flavobacterium (Flavobacterium) bacteria belonging to the genus Shudomomasu (Pseudomonas) bacterium belonging to the genus, for example, Pseudomonas fragi (Pseudomonas f
ragi ) IFO 3458, bacteria belonging to the genus Acinetobacter .
また、本発明においてオレイン酸を添加する培地につ
いてもビール有害菌の培養ないし増殖のために従来より
使用されているものを任意に用いることができる。例え
ば、ブイヨン，ハートインフュージョンブロス,S,BCP,N
BB,GAM,BL,MMなどの既知の培地が用いられる。In the present invention, as the medium to which oleic acid is added, any medium conventionally used for culturing or growing beer harmful bacteria can be used. For example, bouillon, heart infusion broth, S, BCP, N
A known medium such as BB, GAM, BL, MM is used.
培地に対するオレイン酸の添加量は５〜100ppmが適当
であり、20〜60ppmが好適である。ここで、オレイン酸
の添加量（ppm）は１に対して加えたオレイン酸の量
（mg）を意味する。オレイン酸の添加量が100ppmを超え
ても相応する効果が得られない。The amount of oleic acid added to the medium is suitably 5 to 100 ppm, and preferably 20 to 60 ppm. Here, the added amount (ppm) of oleic acid means the amount (mg) of oleic acid added to 1. Even if the added amount of oleic acid exceeds 100 ppm, a corresponding effect cannot be obtained.
上記の条件にてオレイン酸を添加した培地を使用する
と、ビール有害菌の増殖が選択的に促進され、一般細菌
や酵母に対する増殖効果は認められない。なお、オレイ
ン酸を添加したことによるビール有害菌の培養条件に変
化はなく、常法に従って培養を行えばよい。When a medium supplemented with oleic acid under the above conditions is used, the growth of beer harmful bacteria is selectively promoted, and no growth effect on general bacteria and yeast is observed. There is no change in the culture conditions of beer harmful bacteria due to the addition of oleic acid, and the culture may be performed according to a conventional method.
［実施例］ 次に、本発明を試験例および実施例により説明する。Next, the present invention will be described with reference to Test Examples and Examples.
試験例１ ビール有害菌の増殖促進に及ぼすオレイン酸の濃度に
ついて検討した。すなわち、液体NBB培養地にオレイン
酸を各種濃度で加えたものを試験管に分注し、ビール有
害菌としてラクトバチルス・ブレビスAHU 1508およびシ
ュードモナス・フラギIFO 3458を接種し、30℃で19時間
振とう培養し、増殖倍率を求めた。また、参考のため
に、ビール酵母についても同様に試験を行った。結果を
第１表に示す。Test Example 1 The concentration of oleic acid that affects the growth of beer harmful bacteria was examined. That is, oleic acid added at various concentrations to a liquid NBB culture area was dispensed into test tubes, and Lactobacillus brevis AHU 1508 and Pseudomonas flagii IFO 3458 were inoculated as beer harmful bacteria, and shaken at 30 ° C. for 19 hours. After culturing, the growth rate was determined. In addition, a test was similarly performed on brewer's yeast for reference. The results are shown in Table 1.
表から明らかなように、オレイン酸の添加はビール有
害菌の増殖に有効であるが、ビール酵母に対しては、殆
んど効果が認められない。 As is clear from the table, the addition of oleic acid is effective for the growth of beer harmful bacteria, but has little effect on brewer's yeast.
液体MM培地にオレイン酸30ppmを添加したものをＬ字
管に分注し、ビール有害菌としてラクトバチルス・ブレ
ビスAHU 1508,クレブシエラ・アエロゲネスATCC 15050
およびシュードモナス・フラギIFO 3458を接種し、30℃
で24時間振とう培養後の菌数を測定し、世代時間を計算
した。また、参考のため、ビール酵母、枯草菌および大
腸菌についても同様に試験を行った。結果を第２表に示
す。A liquid MM medium supplemented with 30 ppm of oleic acid was dispensed into an L-shaped tube, and Lactobacillus brevis AHU 1508, Klebsiella aerogenes ATCC 15050 was used as a beer harmful bacterium.
And Pseudomonas flagi IFO 3458 at 30 ° C
The number of bacteria after shaking culture for 24 hours was measured, and the generation time was calculated. For reference, brewer's yeast, Bacillus subtilis and Escherichia coli were similarly tested. The results are shown in Table 2.
表から明らかなように、オレイン酸の添加によりビー
ル有害菌の世代時間は大幅に短縮される。しかし、ビー
ル酵母や一般細菌に対しては効果が認められない。 As is evident from the table, the addition of oleic acid significantly reduces the generation time of beer harmful bacteria. However, it has no effect on brewer's yeast or general bacteria.
試験例３ 各種微生物の菌液からメンブランフィルターで集菌
し、これをオレイン酸30ppmを添加したMM培地にて30℃
で19時間培養し、ミクロコロニー20個の直径（μｍ）を
顕微鏡で測定し、該コロニーの平均直径（μｍ）を求め
た。結果を第３表に示す。Test Example 3 Cells were collected from bacterial liquids of various microorganisms using a membrane filter, and the collected cells were then added to an MM medium containing 30 ppm of oleic acid at 30 ° C.
For 19 hours, the diameter (μm) of 20 microcolonies was measured with a microscope, and the average diameter (μm) of the colonies was determined. The results are shown in Table 3.
表から明らかなように、ビール有害菌のコロニーの直
径はオレイン酸添加培地において、約1.7〜1.9倍とな
り、観察が非常に容易になる。一方、ビール酵母や一般
細菌ではオレイン酸添加による影響が認められない。 As is clear from the table, the diameter of the colonies of beer harmful bacteria is about 1.7 to 1.9 times in the medium supplemented with oleic acid, which makes observation very easy. On the other hand, in brewer's yeast and general bacteria, the effect of oleic acid addition is not recognized.
実施例1,比較例１ 第４表に示すように、５種類の培地にオレイン酸を25
ppm加えたもの（実施例）とオレイン酸を加えないもの
（比較例）を調製し、それぞれの培地20mlをシャーレに
流し固化した後、第４表に示す４種のビール有害菌の菌
液（細胞数約100個）を塗布し、これを28℃で培養して
コロニーが目視による検出が可能な日数を測定した。結
果を第４表に示す。Example 1, Comparative Example 1 As shown in Table 4, oleic acid was added to five types of culture media.
A solution containing 4 ppm of beer harmful bacteria shown in Table 4 (Example) and a solution containing no oleic acid (Comparative Example) were prepared, and 20 ml of each medium was poured into a petri dish and solidified. (Approximately 100 cells) were applied and cultured at 28 ° C., and the number of days in which colonies could be visually detected was measured. The results are shown in Table 4.
表から明らかなように、培地にオレイン酸を添加する
ことによりビール有害菌の増殖を促進させることがで
き、ビール有害菌の検出に要する日数を短縮させること
ができる。As is clear from the table, by adding oleic acid to the medium, the growth of beer harmful bacteria can be promoted, and the number of days required for detection of beer harmful bacteria can be shortened.
実施例2,比較例２ ビール有害菌以外の細菌や酵母も増殖可能な培地とし
てＳ培地を選択し、これにオレイン酸を25ppm添加した
ものとオレイン酸を添加しないものを調製し、それぞれ
の培地にビール有害菌（実施例）または他の細菌，酵母
（比較例）を接種して増殖試験を行い、増殖コロニーが
目視可能で最も比較しやすい日数経過後、その増殖状態
を比較した。結果を第５表に示す。オレイン酸無添加の
対照Ｓ培地における菌株の増殖度（コロニーの大きさ）
を基準とし、これと同じ増殖度のものをオレイン酸添加
による増殖効果なし（−と表示）とし、増殖効果の認め
られるものについてはその効果の程度に応じて＋，，
で表わした。 Example 2 and Comparative Example 2 S medium was selected as a medium in which bacteria and yeasts other than beer harmful bacteria can also grow, and a medium to which 25 ppm of oleic acid was added and a medium to which no oleic acid was added were prepared. A beer harmful bacterium (Example) or another bacterium or yeast (Comparative Example) was inoculated into the cultivation test, and a proliferation test was performed. The results are shown in Table 5. Degree of growth of strain in control S medium without oleic acid (colony size)
With the same growth degree as the standard, the growth effect by the addition of oleic acid was regarded as no growth effect (indicated by-), and those with the growth effect were determined according to the degree of the effect.
Indicated by
表から明らかなように、オレイン酸の添加による増殖
効果は酵母に若干認められるものの、細菌には認められ
ず、ビール有害菌に対して優先的に増殖効果を示すこと
が明らかとなった。As is clear from the table, although the growth effect due to the addition of oleic acid was slightly observed in the yeast, it was not observed in the bacteria, and the growth effect was preferentially exerted on harmful beer bacteria.
なお、Ｓ培地の代りにブイヨン,BCP,NBB,ハートイン
フュージョンブロスその他の栄養培地を使用して上記と
同様の増殖効果を調べたところ、オレイン酸の添加によ
る増殖効果はビール有害菌にのみ顕著に認められた。When the same growth effect as above was examined using broth, BCP, NBB, heart infusion broth and other nutrient media instead of the S medium, the growth effect due to the addition of oleic acid was remarkable only for beer harmful bacteria. Was recognized.
［発明の効果］ 本発明の培地を用いれば、ビール有害菌を優先的に増
殖させることができるため、ビール有害菌の有無を短期
間で、かつ正確に判定することが可能である。特に、本
発明の培地はビール有害菌のうちグラム陰性菌について
も迅速に検出することができるという特色を有してい
る。 [Effect of the Invention] Since the harmful beer bacteria can be preferentially grown by using the medium of the present invention, the presence or absence of the harmful beer bacteria can be accurately determined in a short period of time. In particular, the culture medium of the present invention has a feature that gram-negative bacteria among beer harmful bacteria can be rapidly detected.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 国武 直之 静岡県焼津市岡当目10番地 サッポロビ ール株式会社醸造技術研究所内 (72)発明者 鎌田 耕造 愛知県名古屋市千種区千種２丁目16番15 号 サッポロビール株式会社名古屋工場 内 (58)調査した分野(Int.Cl.6，ＤＢ名) C12Q 1/04 C12N 1/20 C12C 11/00 ＢＩＯＳＩＳ（ＤＥＲＷＥＮＴ) ＷＰＩ（ＤＥＲＷＥＮＴ)──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Naoyuki Kunitake 10th Oka Tome, Okame, Yaizu City, Shizuoka Prefecture Inside Sake Brewery Research Institute (72) Inventor Kozo Kamata 2-16-15 Chikusa, Chikusa-ku, Nagoya City, Aichi Prefecture No. Sapporo Beer Co., Ltd. Nagoya Factory (58) Fields surveyed (Int. Cl. 6 , DB name) C12Q 1/04 C12N 1/20 C12C 11/00 BIOSIS (DERWENT) WPI (DERWENT)
Claims (1)
ル有害菌の検出用培地。1. A medium for detecting harmful bacteria of beer containing 5 to 100 ppm of oleic acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63296097A JP2769168B2 (en) | 1988-11-25 | 1988-11-25 | Medium for detecting beer harmful bacteria |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63296097A JP2769168B2 (en) | 1988-11-25 | 1988-11-25 | Medium for detecting beer harmful bacteria |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH02142497A JPH02142497A (en) | 1990-05-31 |
| JP2769168B2 true JP2769168B2 (en) | 1998-06-25 |
Family
ID=17829091
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63296097A Expired - Lifetime JP2769168B2 (en) | 1988-11-25 | 1988-11-25 | Medium for detecting beer harmful bacteria |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2769168B2 (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100753012B1 (en) | 1999-08-03 | 2007-08-30 | 가부시키가이샤 야쿠루트 혼샤 | Fermented milks and their production processes |
| TWI468513B (en) * | 2005-05-27 | 2015-01-11 | Yakult Honsha Kk | Lactic acid bacteria fermentation products and fermented milk foods |
| CN108603163A (en) * | 2015-12-03 | 2018-09-28 | 默克专利股份公司 | The culture medium that chemical composition for growing or detecting microorganism determines |
-
1988
- 1988-11-25 JP JP63296097A patent/JP2769168B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| JPH02142497A (en) | 1990-05-31 |
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