JPH11221070A - Inspection of microorganism and apparatus therefor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for inspecting microorganism, for example, capable of identification of species of bacteria and prediction of their growths in a shortened time and the apparatus therefor, and can previously know the state of specified microorganisms in the specimen after the passage of certain times necessary for proliferation. SOLUTION: The specimen of a microorganism is image-processed and the processed results are compared with the data that are previously stored data to extract the features intrinsic to the microorganism thereby identifying its species, and then calculate the proliferation prediction after the passage of a certain time to previously detect the state of the proliferation of the microorganism in the specimen.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for testing microorganisms and the like and an apparatus therefor.

[0002]

2. Description of the Related Art Normally, in processing foods (fish products such as meats and fish, and all other foods), raw materials to be foods are processed at a factory in a certain form and shipped. In the factory, processed foods are inspected for harmful bacteria. As a conventional method of testing for the presence of bacteria in processed foods, etc.,
There is a method in which a sample is collected, cultured as a chalet (inspection body) in an incubator for a predetermined period of time, and the result of the culture is visually measured and detected by an expert. This is a necessary task. Here, a computer-based method has recently been proposed, which binarizes an image of an inspection object taken into a computer, calculates a determination level, and performs inspection.

However, when capturing this image,
Because it is immediately binarized, if the light source for illuminating and imaging the specimen is different in the irradiation angle, direction, etc. with respect to the specimen, they are immediately grasped. Influences on the results of the inspection may cause an error if the results are extended, and the above-mentioned determination level may not be able to be properly determined. And even if there are bacteria that become toxic bacteria attached, mixed and generated on raw materials that become food,
Many of the poisoning bacteria do not reveal the actual state after a certain period of time, so the raw materials are processed as food in factories, and after the shipment, the bacteria proliferate and reach the consumers' hands. At a stage, it is often the case that poisoning is caused by the bacteria that propagated.

In the conventional method for testing microorganisms, it took a long time to obtain the results of the test. In recent years, direct microscopic methods, membrane dyeing, and the like have been used during the processing process, semi-finished products, and products. Method, impedance method, fluorescent staining method,
Methods such as a fluorescent probe method and a bioluminescence method have been performed, and a quick response has been made possible. However, they have advantages and disadvantages in terms of speed, detection range, economy, and the like, and are not presently satisfactory. In some of them, it is possible to identify the species of a specific bacterium, but in practice, it was impossible to identify the bacterium by most methods.

[0005]

The present invention provides an inspection method and apparatus for an inspection object such as a microorganism, which separates an image of the inspection object cultured in a short time (about 5 to 6 hours) into three primary colors and the like, and On the other hand, calculate the histogram, color, and hue, determine the growth coefficient for predicting the growth of bacterial colonies, and the characteristic coefficient for judging the characteristics unique to each microorganism. By searching and comparing them, the species is identified (identified), and by utilizing their growth factors, etc., after the general culture time (24, 48 hours, etc.) necessary for the bacteria elapses It is an object of the present invention to make it possible to easily know the state after a certain period of time in advance, regardless of a short time, by making a breeding prediction.

[0006]

[Means for Solving the Problems] For example, bacteria (live bacteria)
In order to be able to know in advance the state of reproduction after a certain period of time, such as food, including drinking water, images of test specimens collected from food and processed materials are separated into three primary colors in a computer, and a histogram of each primary color By calculating the hue and color, and comparing them with the data obtained by the above-described method for each bacterium, the characteristics unique to the bacterium are extracted and the type thereof is determined. A method for detecting bacteria and the like, characterized in that a state after a certain period of time necessary for propagation of the specified bacteria is detected in advance by calculating a predicted propagation state, and a method for testing bacteria and the like.

[0007]

Embodiments of the present invention will be described based on examples. FIG. 1 is a flowchart of the apparatus and the inspection method of the present invention, wherein (1) to (12) show a culture apparatus [incubator].
The configuration and operation of (A) are shown. The structure and operation of the incubator will be described in order below. The inoculated medium (agar) is installed at a predetermined location as the incubator (A) and as the Petri dish in (1). When the test body is a liquid, a filter is used to collect the bacteria, and there may be other various forms of the test body.)
Set the culture temperature in (2). This is an incubator (A)
Set the required temperature by operating the operation panel.
Set the culture time in (3). This also sets the required time by operating the operation panel of the incubator (A). (Four)
In the above, the operation panel of the incubator (A) is operated so as to operate in the automatic mode [this is interlocked with the terminal device (B) and the processing device (C) to be described later).

[0008] In (5), the terminal device is notified that it is in the automatic mode (the manual mode is used if necessary when culturing is performed for 24 hours or 48 hours). In the culture time of (6), confirm the automatic mode culture time set in (3). At the set temperature in (7), confirm that it is the culture temperature set in (2). In the temperature adjustment of (8), if the temperature is not within the set temperature range, control is performed to prevent overheating.
In the culture completion notification of (9), the culture completion notification is sent to the terminal device (B).
, And notifies the processing device (C). In the material supply confirmation of (10), it is confirmed whether or not a supply request for the test object is made from the terminal device (B) .In the material absence / end notification of (11),
The terminal (B) is notified that there is no test object, and the automatic mode is terminated.In the material supply and reading in (12), the test object is supplied to the terminal device (B) and a barcode is attached. Read.

(I) to (X) show the configuration and operation of the terminal device (B). In the following, the configuration and operation of those devices will be described in order. In the automatic mode setting in (I), the operation panel of the terminal device (B) is operated to operate the automatic mode [the incubator (A) and the processing device (C) described later).
Linked with]. In the automatic mode notification of (II), the incubator (A) and the processing device (C) are notified of the automatic mode. In the culture completion notification of (III), the system waits until the culture completion notification is sent from the incubator (A), and notifies that the culture completion has been performed. In the material supply request of (IV), the incubator (A) is required to supply a test object. In the material confirmation barcode confirmation of (V), it is confirmed that the inspection object is set at a predetermined position and that the barcode can be read.
(VI) In removing the upper lid, remove the upper lid of the test object. In the confirmation of the material fixed position in (VII), it is confirmed that the inspection object is set at a predetermined position in the terminal device. In the data transmission of (VIII), it is checked whether or not the processing device (C) described later has successfully received the inspection object image set at a predetermined position. In the material discharging operation of (IX), the test object is discharged or accumulated outside the terminal device (B). In the confirmation of the end of (X), it is confirmed whether or not the notification of the absence of the test object has been issued from the incubator (A), and if it has been notified, the end notification is transmitted to the processing device (C).

(A) to (e) show the configuration and operation of the processing unit (C). The configuration and operation will be sequentially described below. (A) is an automatic mode setting, and the operation keys of the processing device (C) are operated to operate the automatic mode [the incubator (A) and the processing device (C) described later). Linked]. (B) Automatic confirmation of each device, incubator (A), terminal device (B)
Is set for each automatic mode. (C)
Is data reception, which receives the inspection object image transmitted from the terminal device (B) and stores it inside. If the data is stored normally, a normal reception signal is output to the terminal device (B).
(D) is data storage, and (E) is a data reception completion notification.

Here, the data transmission (V
The data of the inspection object image transmitted from (III) to the data reception (c) of the processing device (C) is subjected to image processing in the computer. That is, the image is represented by three bytes, and each byte is separated by performing a logical operation. The histogram, hue, and color are calculated for each of the separated data, thereby extracting the characteristics of the microorganism. This characteristic is converted into a coefficient, and a database prepared in advance (data obtained by analyzing and organizing data accumulated in experiments and the like by the same method as described above) is searched, and the bacterial species of the specimen is specified. Further, based on the retrieved data, the growth of bacteria is predicted. Thus, the bacterial species and the number of colonies can be measured in a short culture period.

The series of processing operations of the above-mentioned item 0011 will be described in more detail. In the flowchart of FIG. 2, after the start, (a) captures an image (see below), (b) separates the image into, for example, three primary colors of red; R, green; G, blue; B; The histogram of each primary color is calculated and decomposed into 256 types of density distributions. In (d), the hue and color are calculated, that is, the type and density of the color are determined. At this time, a comparison with the database (D) is performed to determine the bacterial species.
That is, the new data from the image just processed and the database (D) [The configuration of the database includes ID, bacterial species (group / individual, for example, coliform group), and characteristic coefficients (R, G,
B distribution <histogram value>, H, S distribution <hue, color value>, etc. are created by the same method as described above and input in advance. It is extracted and the bacterial species is determined.

Next, (f) extracts the outline of the settlement from the read data, (g) predicts the growth of the settlement, and (h) predicts the number of fungal settlements. And extract the contours of the settlements, and calculate the degree of growth α by the distance between the contours (or the perimeter of each contour or the area of the settlement).
For example, 24 hours and 48 hours later are predicted from the result of calculating [see below and Equation 3]. Further details are shown below [FIGS. 3, 4 (a) to (d) and FIG.
Referring to (a) to (c)], as an example of the image capture stage of FIG. 2A shown in FIG.
The image of the test object (Q ') of the test object (Q) cultured for a certain period of time is taken into a computer (PC) by a camera mechanism (K) as an image reading device.

That is, in the three-primary-color separation stage shown in FIG. 2B, the inspection object image as a memory in the computer (PC) is used.
(Q ') is represented by three bytes of R, G, and B. It is displayed as a multi-valued surface image (color image: an image in which each pixel has a color as well as a gradation), and the three primary colors (basic values of color expression)
Each having 1 [formula; N is the number of brightness, FR is brightness detection point, n represents the luminance, R ₁ is the brightness of the test points, ∩ is a logical product] separated using a histogram calculation formula of ( Tone calculation, that is, density is calculated for each primary color). As this separation method, for example, R, G, and B are overlapped to perform color display. Here, in the color image data, each primary color is represented by a three-digit number (R = 1 digit, G = 10 digits, B = 100). Digit), thereby performing a logical operation on each digit of each primary color to separate each primary color. Next, at the stage of calculating each primary color histogram in FIG. 2C, the histogram of each primary color is calculated by calculating the density distribution for each of R, G, and B as shown in the graph of FIG. 4C. . In the next calculation of hue and color in FIG. 4D, the hue S and hue H are calculated by using the hue and color calculation formula of Expression 2. In this equation, Sr is a hue element of R color, Sg is a hue element of G color, Sb is a hue element of B color, and β
Is a unique coefficient of the image capture system for sharpening the image.

(Equation 1)

(Equation 2)

Next, at the stage of extracting the characteristics of the bacterium shown in FIG. 2 (e), the test object is compared with the data obtained by the above-mentioned method in advance.
Bacterial species that are characteristically common to the test object image (Q ′) of (Q) are extracted. Next, in the outline extraction of the village shown in FIG. 2 (f), an outline based on the read data is extracted. It is Fig.5 (a)
As shown in the figure, the distance (x ₁ ), (y
₁ ) is calculated, and using the growth degree α, prediction (several hours later) of the contours << a >> and << b >> in FIG. 5B is performed. In these figures, the initial contours of bacteria (a) and b) are given by Equation 3 (α
x is a growth degree component in the x-axis direction, αy is a growth degree component in the y-axis direction, E is a growth degree in the x-axis direction, and F is a growth degree in the y-axis direction. Lee '>>, << Bro >>
, And shows a state in which it has expanded into one piece. The number of colonies (the number of colonies) and the like are measured based on the data developed in this manner. It performs numbering (labeling) by scanning in the x and y axis directions as shown in FIG. Next, scanning is performed in the y- and x-axis directions (changing the scanning direction) to determine the number (contour << a >>,
"B" merges into outlines "I '" and "B'",
, Three villages have appeared). This number is the number of settlements, and the number of settlements after a general culture time (24 to 48 hours) is predicted from the growth degree α of the database. That is, in the prediction of the number of settlements, the process of growing the settlements is predicted based on the growth degree α of the database, and the number of settlements is estimated based on the color density (see below) of the settlement surrounding area of the new data and the growth degree. As a result, the bacterial species and the number of settlements are determined. At the same time as the number of settlements is measured by the above method,
Species identification is also performed.

(Equation 3)

FIG. 6 (a) shows the (medium) surface of the test object (Q ₀ ), in which even if bacteria exist, it has not yet clearly appeared. FIG. 6 (b) shows the progress during the culturing, and colonies of colonies (cl) are seen. FIG. 6 (c)
Is the state after elapse of 4 to 6 hours, for example, and this is the culture time. Here, the settlement of the settlement (cl)
With l ₁ ), a color change is seen in the area (w). Therefore, as described above, the inspection object image (Q ′) is read, the bacterial species is identified by comparison with the database (D), and the growth degree, the characteristic coefficient, and the like are set. Next, as a prediction of growth, settlement (cl ₁ )
The growth prediction is performed using the color, hue, color density, growth degree, feature coefficient, and the like of the peripheral area (w). FIG. 6D shows a state in which a further settlement (cl ₂ ) of the settlement (cl ₁ ) in the growth stage and a new settlement (cl ₃ ) are being generated, and FIG.
Then, it is a state in which villages (cl ₂ ) and villages (cl ₃ ) grow further and new villages (cl ₄ ) are generated. FIG. 6F shows, for example, a state after a lapse of 24 hours, and shows that each of the above-mentioned settlements has grown and that many new settlements have occurred.

[0017]

According to the present invention, when inspecting a collected specimen in the processing of food, if a toxic bacterium is found in the specimen, the germ is propagated at any speed. The problem is whether or not the bacteria grow in large numbers, which can naturally cause food poisoning. However, the examination of microorganisms requires a certain period of time, and the time varies depending on various kinds of bacteria, and even bacteria of the same kind are usually not uniform depending on external conditions. The present invention enables a specimen to identify its species and predict its growth degree in a short time in advance, and the state of the identified bacteria after a certain period of time has passed. Can be known in advance in a short time.

[0018] According to the present invention, any person can easily and efficiently inspect the state of propagation of microorganisms and the like in advance without requiring any skill. In other words, the present invention enables microorganisms and the like to be inspected by culturing them in a short period of time (24 hours or 48 hours in the conventional method), and accumulates data on the microorganisms. By comparing, identify microorganisms, predict their occurrence,
In addition to being able to grasp accurately, the inspection accuracy can be enhanced by increasing and accumulating the data to be compared. In this way, the present invention contributes to the thorough management of food safety, which is also required in recent years by PL law (product liability law), HACCP (Ha
zard Analysis Critical Control Point;
Critical control point method) or GMP (Good Manufacturi)
ng Practice; hygiene code) It is very useful when taking measures. Further, the present invention is applied not only to the food field including drinking water, but also to all fields having microorganisms such as medical, hygiene, agriculture, application to soil bacteria, use of seawater, sewage and other water, wood, air use, and the like. The range can be expanded.

[Brief description of the drawings]

FIG. 1 is a flowchart showing an apparatus for microorganisms and the like and an inspection method according to the present invention.

FIG. 2 is a flow chart for microbial image capture, feature extraction, growth prediction, and the like in the present invention.

FIG. 3 is an explanatory view showing a main part of the device of the present invention.

FIG. 4 is a view for explaining the progress of image conversion of a test object according to the present invention.

FIG. 5 is a view for explaining a contour extraction and measurement situation of a settlement of a test object according to the present invention.

FIG. 6 is a diagram for explaining culture and growth prediction of a test body according to the present invention.

[Explanation of symbols]

 (A) Culture device (incubator) (B) Terminal device (C) Processing device (D) Database (Q) Specimen (Q ') Specimen image (K) Camera mechanism (PC) Computer (4) Convex ( 5) Half vapor deposition plate (6) Diffusion plate (7) Surface light emitter (8) Cover (9) Adhesive (10) Double-sided tape (11) Welding end face (12) Welding flash (L) Luminescent lamp (c) Center (D) Slot (e) Mounting protrusion (W) Lead wire (h) Hole

Claims (3)

[Claims] 1. An image processing method for a test object such as a microorganism,
By comparing them with pre-stored data,
By extracting the unique characteristics of the microorganism and determining its type, and then calculating the predicted breeding state after the lapse of a certain period of time, the state after the lapse of a certain period of time necessary for the propagation of the microorganism in the test specimen can be determined in advance. Inspection method for microorganisms, etc., characterized in that they are detected at the same time. 2. An image of a test object obtained from a sample collected from a substance or an object having microorganisms, or an image of the test object which has been subjected to luminescence, staining, or the like before or after culturing if necessary, into three primary colors or the like. By separating, calculating the histogram, hue, and color of each primary color and comparing them with the data obtained by the above-mentioned method such as microorganisms, the characteristics unique to microorganisms are extracted and the type is determined, and then By calculating the predicted breeding state after a certain period of time,
A method for testing microorganisms and the like, characterized in that a state after a certain period of time necessary for propagation of microorganisms in a test body is detected in advance. 3. The method according to claim 2, wherein the inspected specimen is automatically discharged based on an arbitrarily set culturing time, temperature, or the like. Inspecting device for microorganisms etc.