JPS60176598A - Culture medium for cultivating bacteria for detecting acid-producing bacteria - Google Patents

Abstract

PURPOSE:A culture medium for cultivating bacteria, obtained by adding eggshell powder in a culture medium for cultivating the bacteria containing sugar, and capable of judging the presence or absence of an acid produced from dissolution reaction of the eggshell powder by the bacteria. CONSTITUTION:A culture medium for cultivating bacteria, consisting of a culture medium prepared by adding eggshell powder to a culture medium for cultivating the bacteria containing a mixture of sucrose with glucose or either one sugar, and capable of judging the presence or absence of an acid produced from the bacteria by dissolution reaction of the eggshell dissolved by the acid in the culture medium on growing the bacteria capable of decomposing the sugar to produce the acid added thereto.

Description

[Detailed Description of the Invention] Point of the invention: A culture medium is prepared by adding an eggshell method to a bacterial culture medium to which a mixture of sucrose and glucose or either one of them is added. When bacteria that decompose sugar and produce acid are grown in this medium, the eggshells in the medium are dissolved by the produced acid. In other words, the presence or absence of acid produced by bacteria can be determined by the dissolution reaction of this eggshell method.The presence or absence of dissolution of bacteria can be determined using this eggshell method. This can be visually observed by checking whether or not the precipitate disappears using the eggshell method.

Proof 1. A medium having the composition of an experimental method using a solid medium was used. casein peptone 0.3 t,
Sodium chloride 0.5%, potassium phosphate-hydrogen 0.2
5%, 01-5% sucrose and glucose, eggshell method (2,6 by turbidity meter) were added to distilled water at a kernel ratio, and this was used as the basal medium. Since this basal medium is a liquid medium, in order to make it into a solid medium, one layer of agar was added and the same medium was incubated at 121°C for 15 minutes, and then dispensed into petri dishes. The medium was solidified at room temperature to form a solid plate medium.

Treptococcus mutan, a type of streptococcus that decomposes these sugars and produces acid, is placed in this solid medium.
s.

5treptococcus aalivariua,
5treptococcus m1tts and 5t
Reptococcus aanguia 'le was smeared and cultured and grown in an incubator at 37°C. As a result, it was found that both of these bacteria grow on the same medium and ferment the added sugar, and the acid released during this process gradually dissolves the eggshells contained in the medium. did.

The dissolution reaction of this eggshell method could be observed with the naked eye as a ring (band)-shaped clear zone and η were formed around the butterfly colony that had grown and formed on the medium.

When we observed the growth status of the zona pellucida formed by 5Treptococcus mutana, we found that the second day after the start of culture was 6, the 0.5 day was 10, and the 0.7 day was 16.
The area of the diameter of the zona pellucida expanded with the duration of 0-fight and culture time. Furthermore, the results of experiments conducted with other bacterial strains indicate that the formation of the zona pellucida differs depending on the strain.

On the other hand, even when these bacteria were grown in a control medium in which sucrose and glucose were removed from the same medium, no zones of pellucida were formed around the colonies. This is because acid production does not occur due to the absence of sugar. This is because there is no

From the results of this experiment, it is possible to determine the presence or absence and strength of the acid production ability of the strain based on the presence or absence of the lysis reaction of this eggshell method and the size of the lysis reaction.Proof 2. Experiment using a liquid medium In the above basal medium (liquid medium), Streptococcus
usmutana, 5treptococcus 5
alivarius, 5treptococcus
m1tts, and 5treptococcus, sa
When inoculated with C. nguis and continued culturing at 37°C, the eggshell method precipitated in the medium gradually disappears with the passage of culture days. Things became almost unrecognizable.

The disappearance of this eggshell method is due to these five treptococcus
This is the result of the decomposition of sugar in the medium and the reaction of the acid produced during the eggshell method.

On the other hand, in the experimental group in which the shield was removed from the same medium, no disappearance of the precipitate was observed even though these 5treptococcus grew in the medium. This was because there were no acid-derived sugars in the medium. Therefore, from this experimental result, by culturing bacteria in this medium, it becomes possible to select acid-producing bacteria.

Proof 3: Detection of eggshell-dissolving bacteria contained in oral cavity cleaning fluid.Using this medium, an attempt was made to detect acid-producing bacteria living in the oral cavity. That is, first, the inside of the oral cavity was washed for 10 seconds with 10-l physiological saline, and the washing solution was washed 10x.

Diluted 1.02x, 103x, 10'x.

Then, 0.1 microliters of each was placed on the above-mentioned solid plate medium, spread on the surface with a Conlage stick, and cultured in an incubator at 37°C.

As a result, some villages form a pellucid zone, and a pellucid zone forms [2].
The number of villages that formed a clear zone was 102×
The presence of acid-producing bacteria in the oral cavity can be regularly detected using this eggshell undissolved method based on the experimental results of 0. I was able to do that.

Patent applicant: Yuki Kafuji -

Claims (1)

[Claims] A medium that can be added to bacterial culture medium containing sucrose and glucose using the eggshell method.