JP2002355024A - Microbe assaying system - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a simple microbe assaying system enabling the presence disposition, count and the like of aquatic microbes to be assayed by flow injection technique continuously in a maintenance-free condition for a long time and speedily in high sensitivity without being affected by hindering factors. SOLUTION: This microbe assaying system of flow injection design comprises various pipings, various solenoid valves 7, 16, 20 and 29, mixing sections 12 and 25, coiled reaction sections 23 and 31, a thermostat 24, peristaltic pumps 11, 19 and 28, and detective section 32. This system works as follows: a reaction product formed by reaction between microbes and a reaction solution is detected at the detective section 32, the resultant output signal is transmitted as the data into a computer 43 where an arithmetic processing is made to determine the count of the microbes; wherein the solenoid valves 7, 16, 20 and 29 and the peristaltic pumps 11, 19 and 28, etc., are operated under automatic control by a sequencer 45.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to the presence embodiment or the number of microorganisms contained in the sample to be measured (sample solution), or the degree or the like of the microbial contamination of the sample to be measured quickly with high accuracy and low cost it relates microorganism measuring apparatus capable of detecting or measuring.

[0002]

As a method of measuring the number or concentration of microorganisms BACKGROUND ART are contained in the measurement sample liquid, conventionally, to form microbial colonies (colonies) on the agar medium, counting the number of colonies how such measuring the number of microorganisms (hereinafter, referred to as "colony counting method".) it has been widely used by. The colony counting method, for example a dilute solution of the sample itself or 該被 measurement sample to be measured, the microorganism was applied as uniformly distributed on an agar medium, or perform pour the microorganism on a medium containing nutrients Is cultured to grow each microorganism to a colony of a size that can be distinguished, the number of colonies is counted, and the number of microorganisms is obtained from the number of colonies. However, the colony counting method, since generally require 20 hours of incubation time, there is a problem that the measurement of the number of microorganisms quickly and automatically is not possible.

[0003] In recent years, to measure the activity of an enzyme microbial species specifically with microorganisms measurement method to quickly perform counting the number of microorganisms based on the measurement result has been proposed. For example, target microorganisms is a bacterium belonging to Escherichia coli group (hereinafter, abbreviated as "coliform."), The coliform by measuring the hydrolysis characteristics of the beta-D-galactosidase is an enzyme having the coliform bacteria Measurement methods such as counting the number have been proposed.

[0004] Specifically, light absorption by the yellow coloring of the beta-D-galactosidase ortho-nitrophenol produced by enzymatic hydrolysis of the ortho-nitrophenol -β- galactopyranoside elicited by enzymatic catalysis (ONPG) Measurement methods such as detecting luminous intensity, and β
Elicited by enzymatic catalysis of -D- galactosidase, it is a fluorescent enzyme substrate 4-methylumbelliferyl -
beta-D-galactopyranoside (4-MUG) enzymatic hydrolysis produced by 4-methylumbelliferone (4
Measurement method such measuring the fluorescence intensity of -MU) have been proposed. It can measure the number of coliform by measuring the rate of any enzymatic reaction.

[0005] Fluorescence enzyme substrate in the conventional coliforms determination methods using, for example, US as disclosed in Patent No. 5,292,644 or U.S. Pat. No. 5,518,894, etc., to be measured in a sample coliform which are contained in, is held and concentrated by filtration or the like on the filter to which the coliform bacteria can not pass through the filter that holds the coliforms, is contacted with a reagent solution containing a fluorescent enzyme substrate, E. coli by measuring the amount of the decomposed fluorescent substance by an enzyme group holds, it is possible to measure the number of coliform. In this measurement method, so that to improve the detection sensitivity by concentrating to capture and coliforms on the filter.

[0006] The measuring method (analysis method) such for measuring the substance concentration in water using flow injection method is also known. For example, Japanese Patent Application Laid-Open No. 9-90366 discloses a measurement method in which the ammonium ion concentration is quantified by using a flow injection method. In this measurement method, while a reagent solution (sodium hypochlorite) is continuously caused to flow down in a flow channel by injection of a reaction reagent, sample water containing ammonium ions in the flow is driven by a constant flow pump. Continuous feeding by
At the air is continuously charged by the driving of the constant flow pump at the same time, so that the reaction of these in a mixing coil. Then, the reaction product obtained, after separated as a gas by vaporization separator is oxidized to nitric by heating oxidation furnace it. Further, the nitric oxide is detected chemiluminescence intensity generated when reacted with ozone by converting and computing the output of the signal strength, to convert the signal strength ammonium ion concentration.

[0007]

However, for example, US Pat. No. 5,292,644 or US Pat.
In conventional coliforms measuring method disclosed in 518,894 No. etc., for performing the measurement is reacted with a fluorescent enzyme substrate in after being held temporarily concentrated on the filter coliform measurement operation is extremely In addition, there is a problem that the configuration of the measuring device becomes complicated and continuous measurement becomes difficult.

[0008] In the case of using a fluorescent enzyme substrate method, the life of the xenon lamp is generally used as a light source for exciting the fluorescent substance in the detection unit for detecting fluorescence 1
Only about 000 hours. Therefore, when performing continuous measurements, frequently it is necessary to replace the lamp, there is a problem becomes a large load maintenance surface.

[0009] When using flow injection method for the measurement of the number of microorganisms, components contained in the reaction reagent, for example a fluorescent enzyme substrate typically water, such as 4-methylumbelliferyl-beta-D-galactopyranoside since the a sparingly soluble substance, be mixed in a simple mixing section this, it is difficult to dissolve the sample water. Thus hardly occur enzymatic hydrolysis reactions, sensitivity there is a problem decreases.

In addition, fluorescent enzyme substrates are generally weak to heat or high temperatures, while liquids containing high concentrations of surfactants may condense at low temperatures. For this reason, when using a solution in which a fluorescent enzyme substrate and a surfactant are mixed in advance, depending on the environment in which the measurement device is installed, the reaction reagent is degraded and cannot perform a predetermined reaction, and sensitivity is reduced. There is a problem.

Generally, the temperature at which the catalytic enzyme reaction is carried out is 3
0 ° C or higher, but the sample water temperature is lower than 25 ° C
In many cases, the temperature of the sample water must be raised when mixing with the reaction reagent because: However, since the reaction portion not increase suddenly the temperature of the sample water can not immediately carry out the catalytic enzymatic reaction. For this reason, there is a problem that the sensitivity of the microorganism measurement is reduced or the measurement time is lengthened.

[0012] In addition, an enzyme with E. coli group β-D-
Microorganisms in the conventional measurement method to measure the enzymatic hydrolysis characteristics of galactosidase, with beta-D-galactosidase other than coliforms when measuring for example sewage treated water, for example, fungi (including molds) since or filamentous fungi are present in the treated water, when they are present in the water sample, there is a problem measurement errors.

Furthermore, turbidity components that interfere with the measurement of microorganisms may be mixed in the sample water, but measurement of the sample water in which these components are mixed causes measurement errors. In addition, since dirt adheres to the flow cell of the detection unit, it is necessary to frequently correct the sensitivity. Therefore, there is a problem loading maintenance surface increases.

[0014] The present invention was made to solve the conventional problems described above, the presence aspect of microorganisms in the liquid by a flow injection method, when measuring the Suto, continuous maintenance free over a long period of time measurements are possible, without being affected by disturbance factors such as turbidity component, the problem to be objective or solved to provide a simple microorganism measuring apparatus capable of performing fast measurements with high sensitivity.

[0015]

The microorganism measuring apparatus according to the first aspect of the present invention, which has been made to solve the above-mentioned problems, comprises: (i) a pump for pumping a first reaction reagent and a sample solution containing microorganisms; A first mixing unit that mixes by driving, a reaction unit that causes a reaction between the first reaction reagent and the substance of the microorganism in a stage (downstream) downstream of the first mixing unit, and a downstream stage (downstream) of the reaction unit (Ii) a heat retaining means for maintaining the reaction unit at a predetermined temperature, and (iii) a first temperature control means for detecting the reaction product on the side. Control means for controlling the flow of the mixture so that the mixture of the reaction reagent and the sample solution is allowed to stay in the reaction section for a predetermined time and the mixture is sent to the detection section in a predetermined amount. It is assumed that.

[0016] In the microorganism measuring apparatus according to the second aspect of the present invention, the control means controls the flow of the mixture so that a part of the mixture is intermittently (intermittently) or continuously sent to the detection unit. It is characterized in that it is adapted to

In the microorganism measuring apparatus according to a third aspect of the present invention, the first reaction reagent may be a substance that produces a substance that emits fluorescence by the reaction, a buffer solution that keeps the hydrogen ion concentration constant, and a surfactant. It is characterized by containing at least one of them.

[0018] Microorganisms measuring apparatus according to the fourth aspect of the present invention is the reaction unit is characterized in that it is formed by a coiled tube.

The microorganism measuring apparatus according to a fifth aspect of the present invention, the detection unit is a fluorometer, as an excitation light source, and characterized by using any one of an ultraviolet lamp, a black light or xenon flash lamp To do.

In the microorganism measuring apparatus according to a sixth aspect of the present invention, a second mixing section for injecting a second reaction reagent for increasing the sensitivity of the detection section is provided between the reaction section and the detection section. It is characterized by having.

The microorganism measuring apparatus according to a seventh aspect of the present invention, between the reaction section and the detecting section, a second reagent to stop the reaction between the substances in which the first reaction reagent microorganism has Characterized in that a second mixing section for injecting is provided.

The microorganism measuring apparatus according to an eighth aspect of the present invention, in the first mixing section or reaction section, its pipe diameter, and the flow rate of the mixture of the first reaction reagent and the sample liquid, the Reynolds number There 1000 or more (preferably more than 2000)
It is characterized in that the set or controlled to be.

The microorganism measuring apparatus according to a ninth aspect of the present invention, in the reaction section, the flow rate of the mixture of the first reaction reagent and the sample liquid, the Reynolds number at a reaction temperature of 100
0 or more (preferably 2000 or more) is characterized in being set to be.

The microorganism measuring apparatus according to a tenth aspect of the present invention includes a reservoir for temporarily storing the sample solution in the preceding stage of the first mixing section, 該貯 Tomeso and a predetermined temperature of at least one of the first mixing section it is characterized in that the means for holding is provided.

[0025] The microorganism measuring apparatus according to an eleventh aspect of the present invention is characterized in that a vessel keeping means for keeping the vessel holding the first reaction reagent at a predetermined temperature is provided.

The microorganism measuring apparatus according to a twelfth aspect of the present invention, the container for heat insulation means, the temperature of the first reaction reagent 1
And it is characterized in that is adapted to hold the 0 ℃ than 20 ° C. or less.

The microorganism measuring apparatus according to a thirteenth aspect of the present invention, in front of the first mixing section, while passing the microorganisms to be measured, does not pass microorganisms or buoyant particulate matter out of the measurement object first it is characterized in that the first filter portion is provided.

The microorganism measuring apparatus according to a fourteenth aspect of the present invention, in front of the first mixing section, a second filter portion which does not pass through the all microorganisms, and the second filter portion Betsuryuro And a switching valve for switching a flow path between the second filter portion and the piping.

The microorganism measuring apparatus according to a fifteenth aspect of the present invention, in front of the first mixing section, the second filter unit and first filter unit which does not pass through the all microorganisms without separate channel When, it is characterized in that a switching valve for switching the flow path for the first filter portion and second filter portion is provided.

[0030] In the microorganism measuring apparatus according to a sixteenth aspect of the present invention, the reaction in the reaction section is a catalytic enzyme reaction, and the first reaction reagent does not inhibit the catalytic enzyme reaction of the microorganism to be measured. and it is characterized in that it contains an antibiotic to inhibit the catalytic enzymatic reaction of unmeasured microorganisms.

[0031]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiment 1 Hereinafter, Embodiment 1 of the present invention will be described. FIG. 1 shows Embodiment 1 of the present invention.
1 is a system configuration diagram of the microorganism measuring device according to the first embodiment. The microorganism measuring apparatus, using water before disinfection with chlorine or ozone in sewage treatment as the sample water, so as to measure the concentration of coliform water sample. Incidentally, the microorganism measuring apparatus according to the first embodiment, although the main purpose of measuring the concentration of coliform in the sewage, the measurement target is not limited thereto, for example, general non-coliform The present invention is also applicable when measuring the number or concentration of microorganisms.

As shown in FIG. 1, in this organism measuring apparatus, the sample water staying or flow within the tank 1,
Through a pipe 2a and the pipe 2b, and is supplied to the water tank 4 by the pump 3. Here, the water storage tank 4
Has a mechanism for overflowing the sample water, and the excess sample water in the water storage tank 4 is returned to the tank 1 via the pipe 5. As can be understood from Bernoulli's theorem, the internal pressure of the pipes 2a and 2b around the pump 3 changes depending on the height at which the sample water in the tank 1 is collected. Therefore, if the pipe 2b is directly connected to the measuring device main body without providing the water storage tank 4, the sample water flow rate in the measuring device may not be maintained at a preset value. Therefore, in order to place the sample water once under the atmospheric pressure, the water storage tank 4 open to the atmospheric air type is provided.

[0033] sample water in the water tank 4, one end of the pipe 6 (lower end) is inserted, the other end of the pipe 6 (upper end) is connected to the first connection end of the first three-way electromagnetic valve 7 . And
The second connection end of the first three-way solenoid valve 7 is connected to one end of a pipe 8, and the other end of the pipe 8 is connected to an ion-exchanged water storage container 9. The third connection end of the first three-way electromagnetic valve 7,
Peristaltic pump 11 is connected to one end of a pipe 10 which is interposed, the other end of the pipe 10 is connected to the first inlet of the first mixing section 12.

The ion-exchanged water stored in the ion-exchanged water storage container 9 is used when calibrating the sensitivity of the microorganism measuring apparatus and cleaning the inside of the pipe. The first three-way solenoid valve 7
In the normal state where no voltage is applied, the connection is set so that the ab side is connected. However, when the voltage is applied and activated, the ab 'side is connected. First mixing section 12
As usually, T-shaped or Y-shaped three-way pipe is used. Here, an orifice in order to improve the mixing characteristics may be utilized, but in this case the pressure inside the pipe is higher. Therefore, one of the three-way pipe and the orifice may be selected in consideration of the balance between the mixing characteristics of the first mixing unit 12, the torque of the peristaltic pump 11, and the pressure resistance of the piping system.

Further, this microorganism measuring apparatus, a mixed reagent containers 13 and calibration mixed reagent container 14 is provided. Mixed reagent stored in mixed reagent container 13, 4-methylumbelliferyl-beta-D as a fluorescent enzyme substrate
- galactopyranoside (. Hereinafter referred to as "4-MUG") containing, hydrogen ion concentration in the reaction contain a phosphate buffer to hold constant, and to soften the cell membranes of the microorganisms in the cell containing sodium lauryl sulfate as a surfactant to fulfill the role of improving the sensitivity when measuring the fluorescence intensity in roles and subsequent stage makes it possible to react with some β- galactosidase readily 4-MUG enzyme .
One end of a pipe 15 is connected to the mixed reagent container 13, and the other end of the pipe 15 is connected to a first connection end of a second three-way solenoid valve 16.

The calibration mixed reagent container 14 stores a mixture of a fluorescent substance, a buffer and a surfactant for calibration of the microorganism measuring apparatus. Here, 4 as a fluorescent substance
-Methylumbelliferone has been used. The buffer and the surfactant are the same as in the case of the mixed reagent in the mixed reagent container 13. The calibration mixture reagent container 14 at one end of the pipe 17 is connected, the other end of the pipe 17 is connected to the second connecting end of the second three-way solenoid valve 16. Also, the second
Third connecting end of the three-way electromagnetic valve 16, a peristaltic pump 19
There is connected to one end of a pipe 18 which is interposed, the other end of the pipe 18 is connected to the first connection end of the two-way electromagnetic valve 20.
The second three-way electromagnetic valve 16 is in a normal state where no voltage is applied is set so as to communicate the c-d side, when a voltage is actuated is applied communicating c-d 'side.

The two-way solenoid valve 20 is closed in a normal state where no voltage is applied, it is set to open when a voltage is applied. The second connection end of the two-way solenoid valve 20 is connected to one end of the pipe 21, the other end of the pipe 21 is first
It is connected to the second inlet of the mixing section 12. The outlet of the first mixing section 12 is connected to one end of a pipe 22,
The other end is connected to the upstream end of the first reaction unit 23.

[0038] Incidentally, the is a material of the pipes, pipes 2a the peristaltic pump 11, 19 is not interposed, 2b,
6, 8, 15, 17, and 21 may be a general and inexpensive silicon tube or Tygon tube. However,
Piping peristaltic pump 11, 19 are interposed 10,1
8, a dedicated tube having a thickness specified by the peristaltic pump is used. The first reaction part 23 is generally intended a tube consisting of Teflon called fluorine-based resin was molded in a coil shape, most of which are housed in a thermostatic bath 24. The inner diameter of the tube is 1.6 mm.

The reason why the first reaction section 23 is formed into a coil shape in this manner is to save the space of the first reaction section 23 and the thermostatic chamber 24 accommodating the first reaction section 23, and to mix and stir the sample water and the mixed reagent. Is promoted to improve the sensitivity. The thermostat 24 is maintained at a constant temperature. Here, the temperature is set to 37 ° C. because of the measurement of coliform bacteria.
It changes according to the microorganism to be measured. 1st reaction part 2
The downstream end of 3 is connected to the first inlet of the second mixing section 25.

Further, the alkaline solution container 26 is provided in the microorganism measuring apparatus, the alkaline solution is stored for this alkali solution container 26. Here, as the alkaline solution, 2M aqueous sodium hydroxide is used. The alkali solution vessel 26, peristaltic pump 28 is one end of a pipe 27 which is interposed (lower end) is connected, the other end of the pipe 27 (the upper end) is connected to the first connection end of the two-way electromagnetic valve 29 I have. The second connection end of the two-way solenoid valve 29 is connected to one end of the pipe 30, the other end of the pipe 30 and the second mixing unit 3
0 is connected to the second inlet. Two-way solenoid valve 29 is closed in a normal state where no voltage is applied, is set to open when a voltage is applied.

The structure of the second mixing section 25, first mixing section 12
Is the same as Further, as a material of the pipes 27 and 30, a material that can withstand a strong alkali having a hydrogen ion concentration (hereinafter, referred to as “pH”) of 13 or more is used. The upstream end of the second reaction section 31 is connected to the outlet of the second mixing section 25. The second reaction section 31 has a coil-like structure made of Teflon (registered trademark), similar to the first reaction section 23, but may have a length of 0.5 m or less. The second flow cell 33 of the downstream end in the detection portion 32 of the reaction portion 31
Is connected to the upper end.

[0042] In this detection unit 32, the excitation lamp 34 is an excitation light source, a xenon lamp is used. The detection unit 32 includes band-pass filters 35 and 36 (optical filters) that pass only valid wavelengths of the wavelength spectrum of the excitation light and the fluorescence, and a photomultiplier that detects the fluorescence generated in the flow cell 33. A tube 37 is provided. Note that a monochromator may be used instead of the filters 35 and 36. However, since it is not necessary that the filter characteristics be steep and close to a single color, a filter having a large brightness is preferable. The excitation wavelength of the fluorescence is set to 360 nm, and the fluorescence measurement wavelength is 45 nm.
It is set to 0 nm. The lower end of the flow cell 33 is connected one end of a pipe 38, the other end of the pipe 38 is connected to a waste container 39.

The output of the photomultiplier tube 37 is connected to a cable 40
Through is inputted to the AD conversion board 41, where it is converted into a digital signal, it is inputted into the computer 43 through the cable 42 after this. Further, the sequencer 45 is connected to the computer 43 is provided by the RS-232C format via a cable 44. The sequencer 45 is programmed to control the operation / stop of the pump 3, the three-way solenoid valves 7, 16, the two-way solenoid valves 20, 29, and the peristaltic pumps 11, 19, 28. Computer 43 performs the sending of the measurement start trigger to the sequencer 45, plays a role of a condition monitoring and data processing of the sequencer 45.

Hereinafter, the measuring operation of the microorganism measuring apparatus having the above configuration will be described. In this measuring operation, water before disinfection is sucked from the tank 1 by the pump 3 as sample water,
This sample water is temporarily held in the water storage tank 4. The sample water overflowing the water storage tank 4 is supplied to the tank 1 via a pipe 5.
Will be returned to Then, both three-way electromagnetic valve 7 and 16, respectively, and the a-b side and the c-d-side (being or set) switched so as to communicate, two-way solenoid valve 20 is opened, both peristaltic pump 11, 19 Is driven (operates). Thus, the sample water and the mixed reagent are mixed in the first mixing section 1
After being mixed in 2 or the pipe 22, the mixture is sent to the first reaction section 23. Since the sample water from the previous measurement remains in the pipe of the first reaction unit 23, the two peristaltic pumps 11 and 12 are used until the sample water to be measured this time is completely replaced.
19 is driven.

[0045] Both peristaltic pump 11, 19 is stopped when the specimen water last in this way is completely substituted,
Two-way electromagnetic valve 20 is closed. This point is the beginning of an enzyme catalyzed reaction, the holding value of the timer at this point (count value) is set (set) to 0 minutes. In the following, will be the time that this timer is to hold referred to as a "time of reaction-minute". For example, a point one minute after the timer starts counting is referred to as “one minute of reaction”. In the first reaction unit 23, the catalytic hydrolysis reaction of beta-D-galactosidase with the coliform group, 4-MUG is decomposed into galactose and 4-methyl umbelliferone.

Thus, when the timer reaches one minute of the reaction, the two-way solenoid valve 29 is opened while the two-way solenoid valve 20 is closed. And both peristaltic pumps 11 and 28
There is driven, the reaction liquid is fed to the second reaction unit 31 and the flow cell 33. The sodium reaction solution sucked from the alkaline solution container 26 and supplied to the second reaction unit 31 via the second mixing unit 25 causes the second reaction unit to be connected from the first reaction unit 23 via the second mixing unit 25. The reaction solution sent to 31 becomes alkaline, thereby stopping the enzyme-catalyzed reaction.

[0047] Incidentally, the driving time of both peristaltic pump 11, 28 (operation time), the second reaction unit 31 and the flow cell 33
Therefore, only a part of the entire reaction solution in the first reaction unit 23 is sent to the second reaction unit 31 and the flow cell 33. Light emitted from the excitation lamp 34 (xenon lamp) is by the filter 35 becomes excitation light centered intensity near 360 nm, is irradiated to the reaction solution in the flow cell 33. This will excite the 4-methylumbelliferone in the reaction solution, emitted fluorescence in the vicinity of 450nm. This fluorescence is detected by the photomultiplier 37, the output is converted from an analog signal to a digital signal by the AD conversion board 41. Intensity of the converted signal is recorded as data by the computer 43.

The above is the operation of the microorganism measuring apparatus in one minute during the reaction, when the reaction 10 minutes, when the reaction 20 minutes, the reaction 3
The same operation is performed at 0 minutes. That is, two-way solenoid valve 20 is closed, with the two-way solenoid valve 29 is opened, peristaltic pump 11, 28 is driven, the reaction solution sequentially a predetermined amount remaining in the second reaction unit 31 and the flow cell 33 The liquid is sent one by one. The subsequent operation is the same as in the case the reaction 1 minute. The liquid discharged from the flow cell 33 is led to a waste liquid container 39 via a pipe 38,
It is stored here. Because this waste liquid is alkaline,
When disposing, it is necessary to neutralize with hydrochloric acid or the like and discharge it to the sewer.

[0049] To illustrate the effect of microbial measurement device in the above operation. With the above operation, when the reaction 1 minute, when the reaction 10 minutes, when the reaction 20 minutes, the fluorescence intensity when the reaction 30 minutes is automatically recorded in the computer 43. Computer 43
C, BASIC, and performs the automatic operation or computation by a program using a programming language such as FORTRAN. Therefore, 30 minutes 4-
The rate of formation of methylumbelliferone can be calculated. The production rate was originally included in the sample water beta-D
-Proportional to the amount of galactosidase.

[0050] Thus, by observing the rate of formation of 4-methylumbelliferone, a preset calibration curve can be obtained by calculating the number of coliforms. Incidentally, the calibration curve shows the relationship between the conventional coliforms measured by agar culture method and 4-methylumbelliferone formation rate (the calibration resistance), obtained in the past at the collection site of the water sample relevant was based on the data, etc., determine actually measured is desirable.

Embodiment 2 Hereinafter, Embodiment 2 of the present invention will be described. The basic configuration or mechanical configuration of the microorganism measuring apparatus according to the second embodiment is the same as that of the microorganism measuring apparatus according to the first embodiment shown in FIG.
Its operation is different only. Therefore, in order to avoid repetition of the description, the following mainly describes the parts different from the first embodiment.

[0052] In the microorganism measuring apparatus according to the second embodiment, during the operation in a microorganism measuring apparatus according to the first embodiment, when the holding value of the timer is set to 0 minutes, i.e. a time reaction 0 minutes two-way electromagnetic valve 20 is closed when, two-way solenoid valve 29 is opened. Then, peristaltic pump 11, 28 is driven, the reaction solution is sequentially continuously fed to the second reaction unit 31 and the flow cell 33. In Embodiment 1, the reaction solution is fed at the time of one minute of the reaction,
When 0 minutes, when the reaction for 20 minutes, was the intermittent and when the reaction for 30 minutes, in the second embodiment, the reaction liquid is continuously feeding.
Embodiment 2 is different from Embodiment 1 in this point.

[0053] Then, in the second embodiment, the flow rate of the peristaltic pump 11, 28 is set to a value smaller than that of the first embodiment. Specifically, this flow rate is
It is set so that the reaction liquid at the time of the reaction 0 minute remains in the first reaction unit 23 even at the time of 0 minute. Further, operation of detecting the 4-methylumbelliferone in the detection unit 32 continuously. The thus obtained 4-methylumbelliferone formation rate becomes continuous rather than a discontinuous data. Therefore, in the second embodiment, than in the first embodiment, there is an advantage variation error data is reduced.

[0054] Embodiment 3. Hereinafter, a third embodiment of the present invention will be described. The basic structures or the mechanical construction of a microorganism measuring apparatus according to the third embodiment is substantially the same as that of a microorganism measuring apparatus according to the first or second embodiments shown in FIG. 1, the excitation lamps configuration is different only. Therefore, to avoid duplicate description, mainly form 1 or form 2 as different parts of the exemplary embodiment in the following be described.

[0055] In the third embodiment, as the excitation lamp 34, long life UV lamp is used than the xenon lamp. Since the lifetime of the xenon lamp is about 1000 hours, when operating the microorganism measuring apparatus continuously,
Lamps must be replaced frequently. For this reason, it takes time and effort to replace the lamp, and the load of maintenance increases. On the other hand, in the case of a long-life ultraviolet lamp, a life of 10,000 to 20,000 hours can be ensured, so that maintenance work is greatly reduced. Incidentally, the ultraviolet lamp may be one of the specification that there is a peak of sufficient intensity to measure near 360～380Nm, such emit xenon lamp, light in the ultraviolet region of shorter wavelength is required in the third embodiment It is.

[0056] In addition to the UV lamp, black light also can be used. It goes without saying that any other light source that emits light near 360 nm long enough for measurement and has a long life can be used. Since the operation of the microorganism measuring apparatus according to the third embodiment is substantially the same as that of Embodiment 1 or Embodiment 2, the description thereof is omitted here.

[0057] Embodiment 4. Hereinafter, Embodiment 4 of the present invention will be described. Incidentally, the basic configuration or mechanical construction of a microorganism measuring apparatus according to the fourth embodiment is similar to the case of a microorganism measuring apparatus according to the first or second embodiments shown in FIG. 1, its operation or effect Only different. Therefore, to avoid duplicate description, mainly form 1 or form 2 as different parts of the exemplary embodiment in the following be described.

In the microorganism measuring apparatus according to the fourth embodiment, water before disinfection is sucked from the tank 1 as sample water by the pump 3, and the sample water is temporarily stored in the water storage tank 4. Incidentally, the sample water overflowing the water tank 4 is returned to the tank 1 via the pipe 5. Thus, at the time of measurement, both three-way solenoid valves 7 and 16 are, respectively, (set) switching is in the a-b side and the c-d side. The two-way solenoid valve 20 is opened, peristaltic pumps 11 and 19 are driven. The flow rates of the two peristaltic pumps 11 and 19 are set such that the Reynolds number of the reaction solution in the pipe 22 to the first reaction section 23 is 1000 or more.

For example, the inside diameter of the pipe 22 and the first reaction section 23 is 0.6 mm, the flow rate of the peristaltic pump 11 is 20 mL / min, and the flow rate of the peristaltic pump 19 is 3
If a mL / min, since the viscosity coefficient of the temperature 40 ° C. Water is a 0.6592mPa · s, by the following equation 1,
The Reynolds number is about 1000. Reynolds number = (density) × (flow rate) × (tube inner diameter) / (viscosity coefficient) ............ Formula 1

[0060] The flow of liquid in the straight tube, the Reynolds number of 2000 or more, preferably mixed If it is 3,000 or more, but the pipe is turbulent state is shown theoretically, in this case, the sample water and reagent are mixed well, the rate of hydrolysis of the enzymatic hydrolysis reaction is maximized. However, the microorganism measuring apparatus according to the fourth embodiment, the pipe passage from the pipe 22 into the first reaction unit 23 instead of being constituted only by the straight pipe, the first reaction portion 23 is formed into a coil shape because there actually has also stirred and mixed function first reaction part 23. Accordingly, each of the shapes and the Reynolds number of the increase in the first reaction portion 23, increases the stirring and mixing action, by thus improving the stirring and mixing action, the microorganism measuring apparatus is highly sensitive, E. coli There is an advantage that the lower limit for measuring the number of groups is improved.

FIG. 2 is a graph showing the result of an experiment in which the same sample water was subjected to an experimental measurement using only the flow rate of the same sample water by using the microorganism measuring apparatus according to the fourth embodiment. In FIG. 2, the horizontal axis indicates the Reynolds number, but here only the flow rate is changed, so that the value on the horizontal axis is proportional to the flow rate or the flow velocity. The vertical axis indicates the production rate of 4-methylumbelliferone, that is, the β-D-galactosidase activity value, and the higher the value, the higher the sensitivity. According to FIG. 2, it can be seen that the sensitivity with in Reynolds number rises slide into increased.

[0062] To increase the Reynolds number, increase the flow rate of the water sample, either to reduce the tube inner diameter, may be increased water temperature. However, reducing the greatly tube inner diameter much flow, required performance becomes high for both peristaltic pump 11, 19, both peristaltic pump 11, 19
Becomes larger. Moreover, increases and the internal pressure becomes a cause of troubles such as the reaction solution from leaking. Also, 50
Excessive rise in water temperature, such as above ° C, is not preferred because it reduces the enzymatic hydrolysis reaction and in some cases causes the degradation of β-D-galactosidase itself. Therefore,
It is difficult at present to greatly increase the Reynolds number.

Embodiment 5 Hereinafter, with reference to FIG. 3,
Embodiment 5 of the present invention will be described. However, the microorganism measuring apparatus according to the fifth embodiment shown in FIG. 3 has many points in common with the microorganism measuring apparatuses according to the first to third embodiments shown in FIG. Therefore, to avoid duplicate description, the microorganism measuring apparatus shown in FIG. 3 are denoted by the same numbers as in FIG. 1 is the member or component common to the microorganism measuring apparatus shown in FIG. 1, their individual Description is omitted.

[0064] As shown in FIG. 3, the microorganism measuring apparatus according to the fifth embodiment, the interior of the thermostatic chamber 24, the pipe 22 and the first mixing section 12 and the water storage tank 4 is housed. Enzymatic hydrolysis reactions are significantly affected by temperature. Thus, a microorganism measuring apparatus shown in Figure 1, particularly if the water sample in such winter cold, at the time of the start of the measurement the sample water and the mixture reagent was fed into the first reaction unit 23 of the tube water temperature remains still low, enzymatic hydrolysis reaction is significantly lowered.

[0065] Even in this case, the function or action of the thermostatic chamber 24, if after a certain time, the water temperature becomes a predetermined value throughout the first reaction unit 23, the measurement error due to the decrease or seasonal sensitivity Is not preferred because However, this embodiment 5
In this case, since the pipe 22 and the water storage tank 4 are housed in the constant temperature bath 24, there is an advantage that the above-described disadvantage is alleviated and a measurement error is reduced. However, in this case, there is a problem that the size of the thermostat 24 increases and its power consumption increases. The operation of the microorganism measuring apparatus is described in the first to third embodiments.
Since this is the same as the case described above, the description thereof is omitted.

[0066] Embodiment 6. Hereinafter, with reference to FIG. 4,
Embodiment 6 of the present invention will be described. However, the microorganism measuring apparatus according to the sixth embodiment shown in FIG. 4 has many common points with the microorganism measuring apparatuses according to the first to third embodiments shown in FIG. Therefore, to avoid duplicate description, the microorganism measuring apparatus shown in FIG. 4, denoted by the same numbers as in FIG. 1 is the member or component common to the microorganism measuring apparatus shown in FIG. 1, their individual Description is omitted.

As shown in FIG. 4, in the microorganism measuring apparatus according to the sixth embodiment, the mixed reagent container 13 and the calibration mixed reagent container 14 are accommodated in a container thermostat 46. The temperature of the vessel for a constant temperature bath 46, unlike the case of the thermostatic chamber 24 is set at around 10 ° C.. And is 4-methylumbelliferyl-beta-D-galactopyranoside is contained in the mixed reagents used in the sixth embodiment, but resistant to heat than normal fluorescent enzyme substrate, nevertheless the influence of temperature since the received and gradually deteriorated, not preferred that the idea to storage for a long time in high temperature conditions.

[0068] Further, also sodium lauryl sulfate contained in the mixed reagent, in the below 5 ℃ cold conditions such as winter, phenomenon such condense in aqueous solution. And when sodium lauryl sulfate condenses,
There is a possibility that the mixed reagent stored in the mixed reagent container 13 cannot be sucked by the peristaltic pump 19, but this is not preferable. Considering these things, the temperature in the container for the thermostatic bath 46 is preferably on the front and rear 10 to 15 ° C.. By providing the container thermostat 46 in this manner, seasonal measurement errors are reduced,
There is an advantage that troubles such as measurement stoppage are reduced, and maintenance is improved. Note that the operation of the microorganism measuring apparatus is the same as in the first to third embodiments, and a description thereof will be omitted.

Embodiment 7 Hereinafter, referring to FIG.
Embodiment 7 of the present invention will be described. However, the microorganism measuring apparatus according to the seventh embodiment shown in FIG. 5, a microorganism measuring apparatus according to the sixth embodiment shown in FIG. 4 (or microorganisms measuring apparatus shown in FIG. 1) having a lot in common. Therefore, in order to avoid repetition of explanation, in the microorganism measuring apparatus shown in FIG. 5, members or components common to those of the microorganism measuring apparatus shown in FIG.
Their individual description is omitted.

[0070] As shown in FIG. 5, the microorganism measuring apparatus according to the seventh embodiment, in the middle of the pipe 6, the third and fourth two-way electromagnetic valve 47 is interposed. Then, between both three-way electromagnetic valve 47, the pipe respectively 49,5
A first filter 53 and a second filter 54 are inserted through the first filter 53 and the pipes 51 and 52. First filter 53
And second filter 54 are both is desirably those of cartridge which can be easily replaced when the mesh is clogged.

[0071] The first filter 53 is a coarse filter mesh, nominal pore size is about 5 micron. On the other hand, the second filter 54, a fine filter mesh, nominal pore size is about 0.5 microns. The role of the first filter 53 is to remove substances larger than the coliform group to be measured. Also, the second filter 54
Is to eliminate all microorganisms, including coliforms.

Next, the operation of the microorganism measuring apparatus according to the seventh embodiment. However, the basic operation of this microorganism measuring apparatus is the same as that of the sixth embodiment (or the first embodiment), and therefore only different parts will be described. In the seventh embodiment, before the operation according to the sixth embodiment (or the first embodiment) is performed, both the three-way solenoid valves 47 and 48 are switched to the ef side and the gh side, respectively ( Set). The first three-way electromagnetic valve 7 (set) switched to a-b side, peristaltic pump 11 is driven. A series of operations subsequent to this is almost the same as in the case of the sixth embodiment (or the first embodiment), and is hereinafter referred to as a “first operation”.

After the end of the first operation, the two three-way solenoid valves 47 and 48 are switched to the ef 'side and the gh' side, respectively. if it is confirmed to be set to the -b side, peristaltic pump 11 is driven, the same operation is repeated. Below,
This series of operations is referred to as a "second operation". Thus, 4 obtained by the first operation and a second operation
- the difference in the rate of formation of methyl umbelliferone is calculated,
From this difference, using a calibration curve of coliforms which are created in advance, E. coli group count is calculated.

Next, effects obtained by the microorganism measuring apparatus according to the seventh embodiment that operates as described above will be described. In the first operation, a microorganism having a beta-D-galactosidase, such as Candida albicans and fungi, since physically larger than coliforms are removed by the first filter 53. In other words, while the magnitude of coliforms is about 0.5 to 3 microns, Candida is the size of about 5-10 microns, also filamentous fungi or larger 10 micron tangled filaments since to be, Candida and filamentous fungi, the first filter 5 of nominal pore size 5 microns
3 can be easily removed. As described above, microorganisms that cause an error due to a change in the bacterial flora in the sewage can be excluded in advance, so that there is an advantage that a measurement error is reduced.

[0075] Further, the second operation, rather than being included in a microorganism, it is possible to obtain the activity value of beta-D-galactosidase which is dissolved in water. Accordingly, by the first operation with the resulting 4-methylumbelliferone formation rate subtracting the 4-methylumbelliferone formation rates obtained in the second operation, it is possible to remove the influence of background, measurement The error can be made smaller.

Embodiment 8 FIG. Hereinafter, an eighth embodiment of the present invention will be described. The basic configuration or mechanical configuration of the microorganism measuring apparatus according to the eighth embodiment is the same as that of the microorganism measuring apparatus according to the seventh embodiment shown in FIG.
Only the composition of the mixed reagent is different. Therefore, in order to avoid repetition of description, the following mainly describes parts different from the seventh embodiment.

[0077] In the microorganism measuring apparatus according to the eighth embodiment, the components of the mixed solution in advance added to a mixed reagent container 13, 4-MUG, phosphate buffer, addition of a surfactant,
The coliforms include antibiotics that have little effect on activity and inhibit fungal activity. This is different from the seventh embodiment. Specifically, such a antibiotics, amphotericin B, known as anti-fungal agents, is preferably a fluconazole like. By keeping by mixing any of these reagents, that reduce the enzymatic activity, such as candida with beta-D-galactosidase activity in non-coliforms, to remove factors that excessive activity value is evaluated There is an effect that it can be done. Since the operation of the microorganism measuring apparatus are the same as those of the embodiments described above, description thereof is omitted.

[0078]

The microorganism measuring apparatus according to the present invention is configured as described above, and has the following effects. In the flow-injection-type microorganism measuring apparatus according to the first aspect of the present invention including the first mixing section, the reaction section, and the detection section, the reaction section is maintained at a predetermined temperature, and the reaction section has a predetermined temperature. The mixture of the first reaction reagent and the sample solution remaining for a time is sent to the detection unit in a predetermined amount. Therefore, it is possible to quickly detect or measure the existence mode, number, or concentration of microorganisms contained in the sample solution, or the degree of microorganism contamination, with high accuracy and at low cost.

According to the microorganism measuring apparatus according to the second aspect of the present invention, a part of the mixture of the first reaction reagent and the sample liquid remaining in the reaction section for a predetermined time is intermittently or continuously sequentially. since sent to the detection unit, it can be a measurement error smaller.

According to the apparatus for measuring microorganisms according to the third aspect of the present invention, the first reaction reagent is selected from the group consisting of a substance that produces a substance that emits fluorescence, a buffer solution that keeps the hydrogen ion concentration constant, and a surfactant. because it contains at least one of, it may be a measurement error smaller.

According to the microorganism measuring apparatus according to the fourth aspect of the [0081] present invention, since the reaction unit is formed by a coiled tube,
A highly sensitive measurement can be performed and a space-saving device can be provided.

According to the microorganism measuring apparatus according to a fifth aspect of the [0082] present invention, as an excitation light source, an ultraviolet lamp, a black light, so any one of the xenon flash light is used, the load of maintenance is reduced, long life it can be obtained a measuring device.

According to the apparatus for measuring microorganisms according to the sixth aspect of the present invention, the second mixing section for injecting the second reaction reagent for increasing the sensitivity of the detection section is provided between the reaction section and the detection section. Therefore, highly sensitive automatic measurement can be performed.

According to the microorganism measuring apparatus of the seventh aspect of the present invention, the second reaction reagent for stopping the reaction between the reaction reagent and the substance of the microorganism is injected between the reaction section and the detection section. since the mixing portion of the second are provided, it is possible to perform a highly sensitive automated measurements.

According to the microorganism measuring apparatus of the eighth aspect of the present invention, in the first mixing section or the reaction section, the tube diameter and the flow rate of the mixture of the first reaction reagent and the sample liquid are: Reynolds number of 1000 or more (preferably 200
0 or more), it is possible to perform highly sensitive automatic measurement.

According to the microorganism measuring apparatus according to a ninth aspect of the [0086] present invention, in the reaction section, the flow rate of the mixture of the first reaction reagent and the sample solution, over 1000 Reynolds number at the reaction temperature (preferably, since the set to be 2000 or higher), it is possible to perform a highly sensitive automated measurements.

According to the microorganism measuring apparatus according to a tenth aspect of the [0087] present invention, a storage tank for temporarily storing the sample solution in the preceding stage of the first mixing section, at least one of 該貯 Tomeso and the first mixing section Since means for maintaining the temperature at a predetermined temperature is provided, it is possible to perform highly sensitive automatic measurement.

According to the microorganism measuring apparatus according to an eleventh aspect of the [0088] present invention, since the container thermal insulation means for holding a container for holding a first reagent to a predetermined temperature is provided, the load of maintenance is reduced, A long-life device can be obtained.

According to the microorganism measuring apparatus according to a twelfth aspect of the [0089] present invention, since the first reaction reagent is controlled to be kept below 20 ° C. 10 ° C. or more, the load of maintenance is reduced, the length It is possible to provide a device with a long life.

[0090] According to the microorganism measuring apparatus according to a thirteenth aspect of the present invention, while passing the microorganisms to be measured, a first filter portion which does not pass through the microorganism or buoyant particulate material out of the measuring object is provided since less susceptible to fluctuations in the microflora seasonal, it is possible to perform a small measurement with measurement variation errors.

According to the microorganism measuring apparatus according to a fourteenth aspect of the [0091] present invention, a second filter portion which does not pass all microorganisms, a pipe forming the different flow passage and the second filter portion, the second since the switching valve for switching the flow path for the filter section and the pipe is provided, it is possible to perform highly sensitive measurements.

[0092] According to the microorganism measuring apparatus of the fifteenth aspect of the present invention, a second flow path which is separate from the first filter section and does not pass all the microorganisms is provided before the first mixing section.
Is provided, and a switching valve for switching the flow path between the first filter unit and the second filter unit is provided, so that highly sensitive measurement can be performed.

According to the microorganism measuring apparatus according to a sixteenth aspect of the [0093] present invention, the first reaction reagent, it does not inhibit the catalytic enzymatic reaction of microorganisms to be measured, the catalytic enzymatic reaction of microorganisms outside the measurement object because it contains an inhibitory antibiotic, less susceptible to variation of microflora seasonal, it is possible to perform a few measurement with measurement errors.

[Brief description of the drawings]

FIG. 1 is a system configuration diagram of a microorganism measuring apparatus according to Embodiments 1 to 4 of the present invention.

FIG. 2 is a graph showing a relationship between a β-galactosidase activity value and a Reynolds number in a microorganism measuring apparatus according to a fourth embodiment of the present invention.

FIG. 3 is a system configuration diagram of a microorganism measuring apparatus according to a fifth embodiment of the present invention.

FIG. 4 is a system configuration diagram of a microorganism measuring apparatus according to a sixth embodiment of the present invention.

FIG. 5 is a system configuration diagram of a microorganism measuring apparatus according to Embodiments 7 and 8 of the present invention.

[Explanation of symbols]

1 tank, 2a pipe, 2b pipe, 3 a pump,
4 water tank, 5 pipe, 6 a pipe, 7 a first three-way solenoid valve, 8 piping, 9 deionized water storage container,
Reference Signs List 10 piping, 11 peristaltic pump, 12 first mixing section, 13 mixed reagent container, 14 calibration mixed reagent container, 15 piping, 16 second three-way solenoid valve, 17
Pipe, 18 pipe, 19 peristaltic pump, 20
Two-way solenoid valve, 21 pipe, 22 pipe, 23
The first reaction section, 24 a thermostat, 25 second mixing unit,
26 lye container, 27 pipe, 28 a peristaltic pump, 29 two-way solenoid valve, 30 pipe, 31
The second reaction unit, 32 detection unit, 33 a flow cell,
34 excitation lamp, 35 filter, 36 filter, 37 photomultiplier, 38 a pipe, 39 a waste container, 40 cable, 41 AD conversion board,
42 cable, 43 computer, 44 cable, 45 PLC, a constant temperature bath for 46 containers,
47 third three-way electromagnetic valve, 48 a fourth three-way solenoid valve, 4
9 pipe 50 pipe 51 pipe 52 pipe,
53 first filter, 54 a second filter.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) // C12Q 1/02 C12Q 1/02 F term (Reference) 2G045 CB21 FA11 FB01 FB12 GC15 GC22 2G054 AA08 AB03 CE02 EA03 GB02 2G058 AA02 CC12 DA01 DA02 DA07 EB01 EC05 EC07 FA07 GA06 4B029 AA07 BB02 CC01 FA03 FA13 4B063 QA18 QQ06 QQ16 QQ18 QQ35 QR43 QR66 QS02 QS39 QX02

Claims (16)

[Claims] 1. A first mixing section for mixing a first reaction reagent and a sample liquid containing microorganisms by driving a pump, and a substance contained in the first reaction reagent and the microorganisms at a stage subsequent to the first mixing section. In a flow injection type microbial measurement apparatus provided with a reaction section for performing a reaction with the reaction section and a detection section for detecting a product of the reaction at a stage subsequent to the reaction section, Means, and control means for controlling the flow of the mixture so that the mixture of the first reaction reagent and the sample solution is allowed to stay in the reaction section for a predetermined time and the mixture is sent to the detection section in a predetermined amount. A microorganism measuring device, characterized in that it is used. 2. The apparatus according to claim 1, wherein said control means controls a flow of the mixture so that a part of the mixture is intermittently or continuously sent to a detection unit. microorganism measuring apparatus according. Wherein the first reaction reagent substance which produces a substance which emits fluorescence by the reaction, and characterized in that it contains at least one of a buffer and a surfactant to a constant hydrogen ion concentration The microorganism measuring device according to claim 1 or 2, wherein 4. A microorganism measuring apparatus according to the reaction section is any one of claims 1 to 3, characterized in that it is formed by a coiled tube. 5. The apparatus according to claim 1, wherein the detection unit is a fluorometer, and any one of an ultraviolet lamp, a black light, and a xenon flash lamp is used as the excitation light source. microorganism measuring apparatus according. Between 6. reaction unit and the detection unit, according to claim 1 to 5 second mixing unit for injecting a second reagent to improve the sensitivity of the detection unit, characterized in that it is provided Any one
The microorganism measuring device according to any one of the above. 7. A second mixing section is provided between the reaction section and the detection section for injecting a second reaction reagent for stopping a reaction between the first reaction reagent and the substance of the microorganism. microorganism measuring apparatus according to any one of claims 1 to 5, characterized in that there. 8. In the first mixing section or the reaction section, the tube diameter and the flow rate of the mixture of the first reaction reagent and the sample solution are set or controlled such that the Reynolds number becomes 1000 or more. The microorganism measuring device according to any one of claims 1 to 7, wherein: 9. The method according to claim 8, wherein in the reaction section, the flow rate of the mixture of the first reaction reagent and the sample solution is set so that the Reynolds number at the reaction temperature is 1000 or more. Microorganism measuring device. 10. A reservoir for temporarily storing the sample solution in the preceding stage of the first mixing section, that the means for holding at least one of 該貯 Tomeso and first mixing section to a predetermined temperature is provided The microorganism measuring device according to any one of claims 1 to 9, wherein: 11. The microorganism measuring apparatus according to claim 1, further comprising a container heat retaining means for retaining a container holding the first reaction reagent at a predetermined temperature. . 12. A container kept means, microorganism measuring apparatus according to claim 11, the temperature of the first reaction reagent, characterized in that is adapted to hold a 20 ° C. below 10 ° C. or higher. In front of 13. The first mixing section, while passing the microorganisms to be measured, characterized in that the first filter portion which does not pass through the microorganism or buoyant particulate material out of the measuring object is provided microorganism measuring apparatus according to any one of claims 1 to 12,. In front of 14. The first mixing section, a second filter portion which does not pass through the all microorganisms, a pipe forming the different flow passage and the second filter portion, a second filter portion and the any of claims 1 to 13, characterized in that a switching valve for switching the flow path for a pipe is provided 1
The microorganism measuring device according to any one of the above. 15. A second filter section, which forms a separate flow path from the first filter section and does not allow all of the microorganisms to pass therethrough, before the first mixing section, a first filter section, and a second filter. that a switching valve for switching the flow path for the parts is provided a microorganism measuring apparatus according to claim 13, wherein. 16. The reaction in the reaction section is a catalytic enzyme reaction, and the first reaction reagent does not inhibit the catalytic enzyme reaction of the microorganism to be measured, but inhibits the catalytic enzyme reaction of microorganisms not to be measured. microorganism measuring apparatus according to any one of claims 1 to 15, characterized by containing the antibiotic.