JP4886122B2 - Vibrio parahaemolyticus detection medium - Google Patents
Vibrio parahaemolyticus detection medium Download PDFInfo
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- JP4886122B2 JP4886122B2 JP2001165144A JP2001165144A JP4886122B2 JP 4886122 B2 JP4886122 B2 JP 4886122B2 JP 2001165144 A JP2001165144 A JP 2001165144A JP 2001165144 A JP2001165144 A JP 2001165144A JP 4886122 B2 JP4886122 B2 JP 4886122B2
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Description
【０００１】
【発明の属する技術分野】
本発明は、食中毒の原因菌として重要な腸炎ビブリオを高精度かつ簡便に検出することのできる培地及び腸炎ビブリオ検出法に関する。
【０００２】
【従来の技術】
腸炎ビブリオ（Vibrio parahaemolyticus）による食中毒は、日本で年間に発生する細菌性食中毒の大きな割合を占める。腸炎ビブリオは海水中に常在し、魚介類を生食することにより中毒が発生する。この食中毒は、急性胃腸炎であり、発熱、嘔吐、上腹部の腹痛、下痢を特徴とする。
【０００３】
このような食中毒原因菌である腸炎ビブリオの検出培地としては、ＴＣＢＳ（Thiosulfate citrate bile salts sucrose）寒天培地が広く用いられている。このＴＣＢＳ寒天培地は、白糖とpH指示薬を含むものであり、腸炎ビブリオが白糖を利用しないのに対し、ビブリオ属に属し、かつ食中毒の原因菌であるビブリオ・アルギノリティカス（Vibrio alginolyticus）やビブリオ・コレラ（Vibrio cholerae）等が白糖を利用して酸を生成するという性質を利用するものである。すなわち、発育してきた菌は菌体中にpH指示薬を取り込む。従って白糖を利用しない腸炎ビブリオ等はpH指示薬の色調変化がないため濃緑色のコロニーを形成するが、一方、白糖を利用し酸を生成する菌はpH指示薬が黄色に変色するため黄色のコロニーとなる、というものである。
【０００４】
【発明が解決しようとする課題】
しかし、ＴＣＢＳ寒天培地においては、検体中に腸炎ビブリオと上記の類縁菌が共存した場合、腸炎ビブリオの濃緑色のコロニーが他の色に変色してしまうため、正確な検出ができなかった。
従って、本発明の目的は腸炎ビブリオと同じビブリオ属に属する類縁菌が共存しても色の変化等を生じずに、正確かつ簡便に腸炎ビブリオを検出できる培地を提供することにある。
【０００５】
【課題を解決するための手段】
そこで本発明者は、腸炎ビブリオを他の属に属する菌だけでなく、同じ属に属する類縁菌とも正確に区別して検出できる培地組成について種々検討したところ、グラム陽性菌抗菌物質と検出可能な遊離性基を有するβ−グルコシダーゼ基質とを含有する培地を用いれば、グラム陽性菌は生育せず、かつビブリオ・アルギノリティカスやビブリオ・コレラは白色のコロニーを形成し、腸炎ビブリオが選択的にβ−グルコシダーゼ基質由来の特性を有するコロニーを形成するので、腸炎ビブリオが正確かつ簡便に検出できることを見出し、本発明を完成するに至った。
【０００６】
すなわち、本発明は、検出可能な遊離性基を有するβ−グルコシダーゼ基質及びグラム陽性菌抗菌物質を含有する腸炎ビブリオ検出用培地を提供するものである。
また、本発明は上記培地に、検体を接種して培養した後、当該培地上の検出可能なコロニーの有無を検出することを特徴とする腸炎ビブリオの検出法を提供するものである。
【０００７】
【発明の実施の形態】
本発明の培地に用いられる検出可能な遊離性基を有するβ−グルコシダーゼ基質とは、β−グルコシダーゼの作用によって色原体化合物、蛍光性化合物等の検出可能な化合物を培地中に遊離するものであり、例えば色原体化合物又は蛍光性化合物とβ−グルコースとが結合した化合物すなわち、色原体化合物又は蛍光性化合物を遊離し得るβ−グルコピラノシド又はその塩であればよい。腸炎ビブリオの類縁菌であるビブリオ・アルギノリティカスやビブリオ・コレラは、β−グルコシダーゼを分泌しないのに対し、腸炎ビブリオはβ−グルコシダーゼを分泌するので、このβ−グルコシダーゼ基質は、腸炎ビブリオによって分解され、色原体化合物、蛍光性化合物等の検出可能な化合物を遊離するため、腸炎ビブリオは色原体化合物に基づく着色コロニー、蛍光性化合物に基づく蛍光コロニー等の検出可能なコロニーを形成する。具体的なβ−グルコシダーゼ基質としては、５−ブロモ−４−クロロ−３−インドキシル−β−Ｄ−グルコピラノシド（Ｘ−β−グルコシド、青色）、５−ブロモ−６−クロロ−３−インドキシル−β−Ｄ−グルコピラノシド（ＭＡＧＥＮＴＡ−β−グルコピラノシド、赤紫）、５−ブロモ−６−クロロ−３−インドリル−β−Ｄ−グルコピラノシド（赤紫）、６−クロロ−３−インドリル−β−Ｄ−グルコピラノシド（ピンク）、５−ブロモ−３−インドリル−β−Ｄ−グルコピラノシド、４−メチル−ウンベリフェリル−β−Ｄ−グルコピラノシド、ｏ−ニトロフェニル−β−Ｄ−グルコピラノシド、フェニル−β−Ｄ−グルコピラノシド、３−ニトロフェニル−β−Ｄ−グルコピラノシド、４−ニトロフェニル−β−Ｄ−グルコピラノシド、３−インドキシル−β−グルコピラノシドトリハイドレート、ｎ−ヘプティル−β−Ｄ−グルコピラノシド、５−ブロモ−４−クロロ−３−インドキシル−２−アセトアミド−２−デオキシ−β−Ｄ−グルコピラノシド等が挙げられ、このうち鑑別性の点から５−ブロモ−４−クロロ−３−インドキシル−β−Ｄ−グルコピラノシド（Ｘ−β−グルコシド）が好ましい。
【０００８】
かかるβ−グルコシダーゼ基質の培地中の含有量は、基質により異なるが、０．０１〜１０ｇ／Ｌ、特に０．０１〜０．６ｇ／Ｌであるのが検出性の点で好ましい。
【０００９】
グラム陽性菌抗菌物質は、検体中のグラム陽性菌の生育を阻害する目的で培地に添加される。これにより、ビブリオ属等を含むグラム陰性菌のみが生育し得ることとなる。当該グラム陽性菌抗菌物質としては、バンコマイシン、リストセチン、及びこれらの塩等が挙げられ、このうち、培地中における安定性の点からバンコマイシン又はその塩が好ましい。
【００１０】
グラム陽性菌抗菌物質の培地中の含有量は、グラム陽性菌の生育が阻害される量であればよく、用いる抗菌物質の種類によって異なるが、０．０００１〜１ｇ／Ｌ、特に０．００１〜０．１ｇ／Ｌが好ましい。
【００１１】
本発明の培地には、腸炎ビブリオの生育を促進し、他の菌の生育を抑制することによって、さらに正確に腸炎ビブリオを検出する目的で、無機塩類及び／又は糖類を含有させるのが好ましい。ここで無機塩類としては、塩化ナトリウム、チオ硫酸ナトリウム等の無機酸金属塩；クエン酸鉄アンモニウム、クエン酸ナトリウム等の有機酸金属塩等が挙げられる。このうち、塩化ナトリウム等の無機酸金属塩は、腸炎ビブリオが好塩性細菌であるため、生育を促進するうえで特に好ましい。また、他の無機塩類は、胆汁末、コール酸ナトリウム、デオキシコール酸ナトリウム等の胆汁酸類と併用することによりビブリオ属以外のグラム陰性菌の生育を抑制するため好ましい。ここで、塩化ナトリウム等の無機酸金属塩は本発明培地中に１．００〜５０．０ｇ／Ｌ、特に５．００〜３０．０ｇ／Ｌ含有するのが好ましい。有機酸金属塩は、本発明培地中に０．０１〜２０．０ｇ／Ｌ、特に０．５０〜１０．０ｇ／Ｌ含有するのが好ましい。また胆汁酸類は、本発明培地中に０．００１〜５．００ｇ／Ｌ、特に０．１０〜１．００ｇ／Ｌ含有するのが好ましい。
【００１２】
糖類としては、単糖類及びオリゴ糖類が使用でき、例えばラクトース、シュークロース（白糖）、キシロース、セロビオース、マルトース等が挙げられる。これらの糖類の添加により、腸炎ビブリオ以外の菌によって前記β−グルコシダーゼ基質が利用されてしまうのを防止でき、より正確な検出が可能となるため、好ましい。これら糖類は、本発明培地中に１．００〜５０ｇ／Ｌ、特に５〜３０ｇ／Ｌ含有させるのが好ましい。
【００１３】
また、本発明の培地には、ペプトン、酵母エキス、獣肉エキス、魚肉エキス等の栄養成分を含有させるのが好ましい。さらに、ピルビン酸ナトリウム、硫酸塩等を添加することもできる。また、腸炎ビブリオは、アルカリ条件下でよく生育するので、本発明培地にはpHを８．０〜９．０にするためのアルカリ剤を含有するのが好ましい。アルカリ剤としては、炭酸ナトリウム、水酸化ナトリウム、ほう酸ナトリウム、３−［（１，１−ジメチル−２−ヒドロキシエチル）アミノ］−２−ヒドロキシプロパン−スルホン酸、Ｎ，Ｎ−ビス（２−ヒドロキシエチル）グリシン、Ｎ−シクロヘキシル−３−アミノプロパンスルホン酸、シクロヘキシルアミノエタンスルホン酸、Ｎ−トリス（ヒドロキシメチル）メチル−３−アミノ−プロパンスルホン酸等が挙げられる。
【００１４】
本発明培地の形態は特に限定されず、通常の寒天培地の他、シート状簡易培地（例えばメッシュを有する繊維状吸水シートに担持させた構造（特開２０００−３２５０７２））とすることもできる。
【００１５】
本発明培地を用いて、被検体中の腸炎ビブリオを検出するには、当該培地に検体を添加して３０〜４５℃で１８〜２４時間培養した後に、コロニーの着色、蛍光等の特性を観察すればよい。
【００１６】
本発明培地に適用される検体としては、魚介類等の生鮮食料品、海水、調理場などのふき取り検体等が挙げられるが、これらの検体を予めトリプトソーヤブイヨン培地で培養した培養液やさらにこれを増菌用培地で培養した培養液も用いることができる。
【００１７】
【実施例】
次に実施例を挙げて本発明を詳細に説明するが、本発明はこれら実施例に何ら限定されるものではない。
【００１８】
実施例１
ペプトン１０ｇ、酵母エキス５ｇ、塩化ナトリウム１０ｇ、クエン酸ナトリウム１０ｇ、クエン酸鉄アンモニウム１ｇ、チオ硫酸ナトリウム６．４ｇ、白糖末２０ｇ、乳糖１０ｇ、ピルビン酸ナトリウム１ｇ、牛胆汁末０．２５ｇ、Ｘ−β−グルコシド０．３ｇ、寒天１５ｇを１リットルの精製水に加え２０％炭酸ナトリウム水溶液にてpHを８．８に合わせた後、１００℃、２０分間加温溶解する。溶解後培地が冷めたことを確認し、ろ過滅菌したバンコマイシン塩酸塩を０．０２ｇ／Ｌになるように加え良く撹拌した後、プラスチックシャーレ（９０φmm）に２０mLずつ分注して培地が固まるまで静置し、本発明の腸炎ビブリオ検出用培地を作製した。
【００１９】
試験例１
供試菌株はトリプトソイブイヨン（腸炎ビブリオ及びビブリオ・アルギノリティカスは３％食塩加トリプトソイブイヨン）で２４時間前培養したものを用い、これを滅菌生理食塩水（腸炎ビブリオ及びビブリオ・アルギノリティカスは滅菌３％食塩水）で希釈し本発明の腸炎ビブリオ検出用培地に接種した。
接種後３５℃で２４時間培養したところ、表１に示すように本発明培地上では腸炎ビブリオは青色の発色コロニーを形成し、ビブリオ・アルギノリティカス、ビブリオ・コレラ等は白色のコロニーを形成し、グラム陽性球菌はその発育が阻止された。これにより、本発明培地を用いれば、腸炎ビブリオが正確かつ簡便に検出できることがわかる。
【００２０】
【表１】
実施例２
実施例１の組成の培地成分を用いて、メッシュを有する繊維状吸水シートに担持させた簡易培地（特開２０００−３２５０７２公報）実施例１（ヒドロキシプロピルセルロース０．１重量％におけるブドウ糖２ｇ／Ｌとテトラゾリウムクロライド０．０２ｇ／Ｌを塩化ナトリウム１０ｇ／Ｌ、クエン酸鉄アンモニウム１ｇ／Ｌ、チオ硫酸ナトリウム６．４ｇ／Ｌ、白糖末２０ｇ／Ｌ、乳糖１０ｇ／Ｌ、ピルビン酸ナトリウム１ｇ／Ｌ、牛胆汁末０．２５ｇ／Ｌ、Ｘ−β−グルコシド０．３ｇ／Ｌに代えて調製）に試験例１と同様にして供試菌株を適用した。その結果、表２に示すように本発明簡易培地によっても、腸炎ビブリオが正確かつ簡便に検出できた。なお、表２には実施例１の寒天培地による結果も併せて示した。
【００２２】
【表２】
【発明の効果】
本発明培地を用いれば食中毒原因菌として重要な腸炎ビブリオが正確かつ簡便に検出できる。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a medium and a method for detecting Vibrio parahaemolyticus that can detect Vibrio parahaemolyticus important as a causative agent of food poisoning with high accuracy and ease.
[0002]
[Prior art]
Food poisoning caused by Vibrio parahaemolyticus accounts for a large proportion of bacterial food poisoning that occurs annually in Japan. Vibrio parahaemolyticus is resident in seawater, and poisoning occurs by eating seafood. This food poisoning is acute gastroenteritis and is characterized by fever, vomiting, upper abdominal pain, and diarrhea.
[0003]
A TCBS (Thiosulfate citrate bile salts sucrose) agar medium is widely used as a detection medium for Vibrio parahaemolyticus which is a food poisoning causative bacterium. This TCBS agar medium contains sucrose and a pH indicator, whereas Vibrio parahaemolyticus does not use sucrose, whereas Vibrio alginolyticus ( Vibrio alginolyticus ) and Vibrio which belong to the genus Vibrio and cause food poisoning.・ Cholera ( Vibrio cholerae ), etc. uses the property of producing acid using sucrose. That is, the growing bacteria take up a pH indicator into the cells. Therefore, Vibrio parahaemolyticus, etc. that does not use sucrose forms dark green colonies because there is no change in the color tone of the pH indicator, whereas bacteria that produce acid using sucrose change to yellow colonies because the pH indicator turns yellow. It will be.
[0004]
[Problems to be solved by the invention]
However, in the TCBS agar medium, when Vibrio parahaemolyticus and the above-mentioned related bacteria coexist in the specimen, the dark green colonies of Vibrio parahaemolyticus discolor to other colors, so that accurate detection could not be performed.
Accordingly, an object of the present invention is to provide a medium capable of accurately and simply detecting Vibrio parahaemolyticus without causing a color change or the like even when related bacteria belonging to the same Vibrio genus as Vibrio parahaemolyticus coexist.
[0005]
[Means for Solving the Problems]
Therefore, the present inventor has conducted various studies on medium compositions that can accurately detect Vibrio parahaemolyticus not only from bacteria belonging to other genera but also from related bacteria belonging to the same genus. If a medium containing a β-glucosidase substrate having a sex group is used, Gram-positive bacteria do not grow, Vibrio arginoliticus and Vibrio cholera form white colonies, and Vibrio parahaemolyticus is selectively β -Since a colony having characteristics derived from a glucosidase substrate is formed, it was found that Vibrio parahaemolyticus can be detected accurately and simply, and the present invention has been completed.
[0006]
That is, the present invention provides a medium for detecting Vibrio parahaemolyticus containing a β-glucosidase substrate having a detectable free group and an antibacterial substance of Gram-positive bacteria.
The present invention also provides a method for detecting Vibrio parahaemolyticus characterized by detecting the presence or absence of detectable colonies on the medium after inoculating the sample with the specimen and culturing.
[0007]
DETAILED DESCRIPTION OF THE INVENTION
The β-glucosidase substrate having a detectable free group used in the culture medium of the present invention releases a detectable compound such as a chromogenic compound or a fluorescent compound into the medium by the action of β-glucosidase. Yes, for example, a compound in which a chromogen compound or a fluorescent compound and β-glucose are combined, that is, a β-glucopyranoside or a salt thereof that can liberate the chromogen compound or the fluorescent compound. Vibrio arginolyticus and Vibrio cholera, which are related to Vibrio parahaemolyticus, do not secrete β-glucosidase, whereas Vibrio parahaemolyticus secretes β-glucosidase, so this β-glucosidase substrate is degraded by Vibrio parahaemolyticus. In order to release detectable compounds such as chromogenic compounds and fluorescent compounds, Vibrio parahaemolyticus forms detectable colonies such as colored colonies based on chromogenic compounds and fluorescent colonies based on fluorescent compounds. Specific β-glucosidase substrates include 5-bromo-4-chloro-3-indoxyl-β-D-glucopyranoside (X-β-glucoside, blue), 5-bromo-6-chloro-3-indoxyl. -Β-D-glucopyranoside (MAGENTA-β-glucopyranoside, red purple), 5-bromo-6-chloro-3-indolyl-β-D-glucopyranoside (red purple), 6-chloro-3-indolyl-β-D -Glucopyranoside (pink), 5-bromo-3-indolyl-β-D-glucopyranoside, 4-methyl-umbelliferyl-β-D-glucopyranoside, o-nitrophenyl-β-D-glucopyranoside, phenyl-β-D -Glucopyranoside, 3-nitrophenyl-β-D-glucopyranoside, 4-nitrophenyl-β-D-glucopyranoside, 3 And indoxyl-β-glucopyranoside trihydrate, n-heptyl-β-D-glucopyranoside, 5-bromo-4-chloro-3-indoxyl-2-acetamido-2-deoxy-β-D-glucopyranoside, and the like. Of these, 5-bromo-4-chloro-3-indoxyl-β-D-glucopyranoside (X-β-glucoside) is preferable from the viewpoint of distinguishability.
[0008]
The content of the β-glucosidase substrate in the medium varies depending on the substrate, but is preferably 0.01 to 10 g / L, particularly 0.01 to 0.6 g / L from the viewpoint of detectability.
[0009]
The antibacterial substance of gram positive bacteria is added to the medium for the purpose of inhibiting the growth of gram positive bacteria in the specimen. As a result, only gram-negative bacteria including Vibrio can be grown. Examples of the antibacterial substance of Gram-positive bacteria include vancomycin, ristocetin, and salts thereof, among which vancomycin or a salt thereof is preferable from the viewpoint of stability in the medium.
[0010]
The content of the antibacterial substance of Gram-positive bacteria in the medium may be an amount that inhibits the growth of Gram-positive bacteria, and varies depending on the type of antibacterial substance used, but is 0.0001-1 g / L, especially 0.001- 0.1 g / L is preferred.
[0011]
The medium of the present invention preferably contains inorganic salts and / or saccharides for the purpose of more accurately detecting Vibrio parahaemolyticus by promoting the growth of Vibrio parahaemolyticus and suppressing the growth of other bacteria. Examples of inorganic salts include inorganic acid metal salts such as sodium chloride and sodium thiosulfate; organic acid metal salts such as ammonium iron citrate and sodium citrate. Of these, inorganic acid metal salts such as sodium chloride are particularly preferable for promoting growth because Vibrio parahaemolyticus is a halophilic bacterium. In addition, other inorganic salts are preferable because they inhibit the growth of gram-negative bacteria other than Vibrio spp. By using them together with bile acids such as bile powder, sodium cholate and sodium deoxycholate. Here, the inorganic acid metal salt such as sodium chloride is preferably contained in the medium of the present invention in an amount of 1.00 to 50.0 g / L, particularly 5.00 to 30.0 g / L. The organic acid metal salt is preferably contained in the medium of the present invention in an amount of 0.01 to 20.0 g / L, particularly 0.50 to 10.0 g / L. Bile acids are preferably contained in the culture medium of the present invention in an amount of 0.001 to 5.00 g / L, particularly 0.10 to 1.00 g / L.
[0012]
As the saccharide, monosaccharides and oligosaccharides can be used, and examples thereof include lactose, sucrose (sucrose), xylose, cellobiose, and maltose. The addition of these saccharides is preferable because it can prevent the β-glucosidase substrate from being used by bacteria other than Vibrio parahaemolyticus and enables more accurate detection. These saccharides are preferably contained in the medium of the present invention at 1.00 to 50 g / L, particularly 5 to 30 g / L.
[0013]
The medium of the present invention preferably contains nutrient components such as peptone, yeast extract, animal extract, fish extract and the like. Further, sodium pyruvate, sulfate and the like can be added. Moreover, since Vibrio parahaemolyticus grows well under alkaline conditions, it is preferable that the culture medium of the present invention contains an alkaline agent for adjusting the pH to 8.0 to 9.0. Examples of the alkali agent include sodium carbonate, sodium hydroxide, sodium borate, 3-[(1,1-dimethyl-2-hydroxyethyl) amino] -2-hydroxypropane-sulfonic acid, N, N-bis (2-hydroxy And ethyl) glycine, N-cyclohexyl-3-aminopropanesulfonic acid, cyclohexylaminoethanesulfonic acid, N-tris (hydroxymethyl) methyl-3-amino-propanesulfonic acid, and the like.
[0014]
The form of the medium of the present invention is not particularly limited, and may be a sheet-like simple medium (for example, a structure supported on a fibrous water-absorbing sheet having a mesh (Japanese Patent Laid-Open No. 2000-325072)) in addition to a normal agar medium.
[0015]
In order to detect Vibrio parahaemolyticus in a subject using the culture medium of the present invention, the specimen is added to the medium and cultured at 30 to 45 ° C. for 18 to 24 hours, followed by observation of characteristics such as colony coloration and fluorescence. do it.
[0016]
Samples applied to the culture medium of the present invention include fresh foods such as seafood, seawater, wiping samples such as kitchens, etc., but these samples can be cultured in advance in a tryptosoya bouillon medium. A culture solution obtained by culturing this in a medium for enrichment can also be used.
[0017]
【Example】
EXAMPLES Next, although an Example is given and this invention is demonstrated in detail, this invention is not limited to these Examples at all.
[0018]
Example 1
Peptone 10 g, yeast extract 5 g, sodium chloride 10 g, sodium citrate 10 g, ammonium iron citrate 1 g, sodium thiosulfate 6.4 g, sucrose powder 20 g, lactose 10 g, sodium pyruvate 1 g, bovine bile powder 0.25 g, X- Add 0.3 g of β-glucoside and 15 g of agar to 1 liter of purified water, adjust the pH to 8.8 with 20% aqueous sodium carbonate solution, and dissolve by heating at 100 ° C. for 20 minutes. After dissolution, confirm that the medium was cooled, add vancomycin hydrochloride sterilized by filtration to 0.02 g / L and stir well, then dispense 20 mL each into a plastic petri dish (90 mm), and let stand until the medium solidifies. The medium for detecting Vibrio parahaemolyticus of the present invention was prepared.
[0019]
Test example 1
The test strain used was pre-cultured for 24 hours in Tryptosoy bouillon (Vibrio parahaemolyticus and Vibrio arginoliticus was 3% saline tryptic soy bouillon), and this was sterilized with physiological saline (Vibrio parahaemolyticus and Vibrio arginolyticus). Was diluted with sterile 3% saline) and inoculated into the Vibrio parahaemolyticus detection medium of the present invention.
When inoculated at 35 ° C. for 24 hours after inoculation, Vibrio parahaemolyticus forms a blue colored colony and Vibrio arginolyticus, Vibrio cholera and the like form white colonies on the medium of the present invention as shown in Table 1. Gram-positive cocci were prevented from growing. This shows that Vibrio parahaemolyticus can be detected accurately and simply by using the culture medium of the present invention.
[0020]
[Table 1]
Example 2
Example 1 (Glucose 2 g / L in 0.1% by weight of hydroxypropyl cellulose) Simple medium (Japanese Patent Laid-Open No. 2000-325072) supported on a fibrous water-absorbing sheet having a mesh using the medium components having the composition of Example 1 And tetrazolium chloride 0.02 g / L, sodium chloride 10 g / L, ammonium iron citrate 1 g / L, sodium thiosulfate 6.4 g / L, sucrose powder 20 g / L, lactose 10 g / L, sodium pyruvate 1 g / L, The test strain was applied in the same manner as in Test Example 1 to 0.25 g / L of bovine bile powder and prepared in place of 0.3 g / L of X-β-glucoside. As a result, as shown in Table 2, Vibrio parahaemolyticus could be detected accurately and simply by the simple medium of the present invention. Table 2 also shows the results of the agar medium of Example 1.
[0022]
[Table 2]
【Effect of the invention】
If the culture medium of the present invention is used, Vibrio parahaemolyticus important as a food poisoning bacterium can be detected accurately and simply.
Claims (3)
（Ａ）検出可能な色原体化合物又は蛍光性化合物を遊離し得るβ−グルコピラノシド又はその塩を有するβ−グルコシダーゼ基質、及び
（Ｂ）バンコマイシン、リストセチン及びこれらの塩から選ばれるグラム陽性菌抗菌物質
（Ｃ）塩化ナトリウム及びチオ硫酸ナトリウム
（Ｄ）クエン酸ナトリウム又はクエン酸鉄アンモニウム
（Ｅ）胆汁酸類
を含有する腸炎ビブリオ検出用培地。The following components (A), (B), (C) , ( D) and (E)
(A) a β-glucosidase substrate having β -glucopyranoside or a salt thereof capable of releasing a detectable chromogenic compound or fluorescent compound , and (B) a gram-positive bacterial antibacterial substance selected from vancomycin, ristocetin and salts thereof (C) Sodium chloride and sodium thiosulfate (D) Sodium citrate or iron ammonium citrate
(E) A medium for detecting Vibrio parahaemolyticus containing bile acids.
［化合物群α］
５−ブロモ−４−クロロ−３−インドキシル−β−Ｄ−グルコピラノシド、５−ブロモ−６−クロロ−３−インドキシル−β−Ｄ−グルコピラノシド、５−ブロモ−６−クロロ−３−インドリル−β−Ｄ−グルコピラノシド、６−クロロ−３−インドリル−β−Ｄ−グルコピラノシド、５−ブロモ−３−インドリル−β−Ｄ−グルコピラノシド、４−メチル−ウンベリフェリル−β−Ｄ−グルコピラノシド、ｏ−ニトロフェニル−β−Ｄ−グルコピラノシド、フェニル−β−Ｄ−グルコピラノシド、３−ニトロフェニル−β−Ｄ−グルコピラノシド、４−ニトロフェニル−β−Ｄ−グルコピラノシド、３−インドキシル−β−グルコピラノシドトリハイドレート、ｎ−ヘプティル−β−Ｄ−グルコピラノシド、５−ブロモ−４−クロロ−３−インドキシル−２−アセトアミド−２−デオキシ−β−Ｄ−グルコピラノシドThe medium for detecting Vibrio parahaemolyticus according to claim 1 or 2 , wherein β-glucopyranoside or a salt thereof capable of releasing a chromogenic compound or a fluorescent compound is selected from the following compound group α.
[Compound group α]
5-bromo-4-chloro-3-indoxyl-β-D-glucopyranoside, 5-bromo-6-chloro-3-indoxyl-β-D-glucopyranoside, 5-bromo-6-chloro-3-indolyl- β-D-glucopyranoside, 6-chloro-3-indolyl-β-D-glucopyranoside, 5-bromo-3-indolyl-β-D-glucopyranoside, 4-methyl-umbelliferyl-β-D-glucopyranoside, o- Nitrophenyl-β-D-glucopyranoside, phenyl-β-D-glucopyranoside, 3-nitrophenyl-β-D-glucopyranoside, 4-nitrophenyl-β-D-glucopyranoside, 3-indoxyl-β-glucopyranoside trihydrate N-heptyl-β-D-glucopyranoside, 5-bromo-4-chloro-3-indoxy -2-acetamido-2-deoxy-beta-D-glucopyranoside
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