WO2004021282A1

WO2004021282A1 - Animal behavior analysis method, animal behavior analysis system, animal behavior analysis program, and computer-readable recorded medium on which the program is recorded

Info

Publication number: WO2004021282A1
Application number: PCT/JP2003/010979
Authority: WO
Inventors: Satoru Kato; Ken-Ichiro Muramoto
Original assignee: Japan Science And Technology Agency
Priority date: 2002-08-29
Filing date: 2003-08-28
Publication date: 2004-03-11
Also published as: JP2004089027A

Abstract

A behavior analysis system comprises a CCD camera for capturing images of a zebrafish in an aquarium at a frame rate of 8 images/sec and a general measurement unit for acquiring observation data from the images. A pursuing behavior evaluating unit evaluates the pursuing behavior of a zebrafish pursuing another zebrafish with reference to the position and speed of the zebrafish, which are observation data on the zebrafish. A fractal dimension calculating unit calculates the fractal dimension of a swim track from the measured data on the zebra fish, which is observation data. Thus, the analysis of behavior of a zebrafish, which is most suitable as a laboratory animal for gene research and swims fast, can be made quantitatively and accurately.

Description

Description Animal behavior analysis method, animal behavior analysis system, animal behavior analysis program, and computer-readable recording medium recording the program

The present invention relates to an animal behavior analysis method and an animal behavior analysis system, and more particularly, to an animal behavior analysis method for quantitatively analyzing animal behavior, an animal behavior analysis system, an animal behavior analysis program, and The present invention relates to a computer-readable recording medium on which the information is recorded. Background art

Recent progress in gene research has been remarkable, and the research subjects have shifted from lower creatures such as nematodes, fruit flies, and yeast to higher animals such as mice, fish, and humans. In gene research, analysis of all genomes has been completed: the boss genome has entered the era of proteomics, which analyzes the function of the protein (protein) encoded by the genome. It is anticipated that we will move into the phenomena of analyzing phenotypes.

Here, it has been widely recognized that zebrafins are the most suitable animal species for genetic modification and gene knockout due to their extremely advantageous features in developmental biology, and in recent years, genomic It is one of the most advanced animals analyzed.

More specifically, zeprafish is a small tropical fish with a length of 5 to 6 cm, and unlike other fish, it lays eggs at any time of the year. (2) Since the juvenile body is transparent, gene expression can be tracked and observed from outside using a fluorescent protein such as GFP (red fluorescent protein). are doing.

In zebrafish gene research, analysis of the whole genome is imminent, and in the future, developmental biology, fish behavior analysis as a phenotype, that is, genetic modification and gene transfer The demand for behavioral analysis of fish subjected to knockouts is immense.

However, behavioral analyzers for small animals such as rats and mice are often used for medical research and are commercially available, but few are for fish at present.

In addition, the inventors described Kato, S., Tamada, K., Shimada, Υ. And Chujo, T., A quantification of goldfish behavior by an image processing system, Behavioral Brain Research, 80 (1996) 51-55. In this study, we proposed an image processing system for analyzing the behavior of goldfish, which captures goldfish in an aquarium and analyzes the position distribution and direction of travel of the goldfish. The inventors have used this image processing system to analyze the behavior of goldfish, particularly quantitative analysis of abnormal behavior after optic nerve transection.

In addition, the Japanese published patent publication “Japanese Patent Application Laid-Open No. Hei 7-63 747 (published on: March 10, 1995)” identifies the activities and survival of fish schools in aquariums. Accordingly, a “water quality inspection device” for detecting an abnormality in water quality is described. This water quality inspection device takes an image of the fish school released into the aquarium with an ITV camera, and executes sampling and binarization processing with the image input unit. By setting the threshold value for binarization, the influence of disturbance noise can be eliminated. The image processing unit converts the binarized image It extracts the images of the individual fish that make up the fish school from the data, attaches a label to each image, and identifies the activity status and survival status of the fish school based on the amount of movement per unit time.

As described above, there is no appropriate behavior analysis method or behavior analysis system, although quantitative analysis of the behavior of zebrafish is necessary and urgent. In other words, in the behavior analysis system and the water quality inspection device for goldfish, zebrafish perform high-speed and complicated three-dimensional behavior, so that quantification is precise enough to meet the needs of gene research. Objective behavior analysis was not possible.

Disclosure of the invention

An object of the present invention is to provide an animal behavior analysis method and an animal behavior analysis system capable of performing quantitative behavior analysis as precisely as possible to meet the requirements of gene research. Further, an object of the present invention includes providing an animal behavior analysis program for realizing the above-described animal behavior analysis system, and a computer-readable recording medium recording the program.

In order to achieve the above object, an animal behavior analysis method of the present invention comprises: an imaging step for imaging an aquatic animal in an aquarium at a frame rate of 8 or more per second; and an image captured in the imaging step. And an analysis step for analyzing the behavior of aquatic animals based on.

Further, the animal behavior analysis method of the present invention is characterized in that the aquatic animal is zebrafish.

Also, the animal behavior analysis system of the present invention captures aquatic animals in an aquarium. An animal behavior analysis system for analyzing the behavior of the aquatic animal based on the obtained image, characterized by comprising imaging means for imaging the image at a frame rate of eight or more seconds. ing.

With the above method and configuration, by capturing images that serve as the basis for behavior analysis at a frame rate of 8 or more Z-seconds, behavior analysis of aquatic animals that perform fast and complex behavior becomes possible.

As explained in the background art, zebrafish has extremely advantageous features in terms of its developmental biology, such as behavioral analysis as a phenotype, that is, genetic modification and genetic modification. There is a strong demand for the establishment of a technique for quantitatively analyzing the behavior of a zebrafish that has been subjected to a knockout.

Therefore, according to the above-described method and configuration, even aquatic animals such as zebrafish that perform high-speed and complicated three-dimensional behavior can be imaged with certainty. Become. Therefore, it is possible to analyze the behavior of zebrafish precisely and quantitatively to the extent that it can meet the needs of gene research.

Further, the animal behavior analysis method of the present invention includes an observation data acquisition step of acquiring the positions and velocities of the first animal and the second animal; and the first animal and the second animal acquired in the observation data acquisition step. And a tracking action evaluation step for evaluating a tracking action of the first animal with respect to the second animal based on the position and velocity of the second animal.

Further, the animal behavior analysis system of the present invention comprises: observation data acquisition means for acquiring the position and velocity of the first animal and the second animal; and the first behavior data acquired by the observation data acquisition means. A tracking line that evaluates the tracking behavior of the first animal with the second animal based on the position and velocity of the animal and the second animal And dynamic evaluation means.

According to the above method and configuration, the tracking behavior of the animal can be quantitatively evaluated based on the position and the speed. Note that the first animal and the second animal may be of the same species or different species. In addition, the first animal and the second animal may be aquatic animals such as zebrafish or resident animals such as Drosophila and mouse.

Here, specific evaluation methods of the tracking behavior include, for example, the condition of the distance between the first animal and the second animal (distance between individuals), the progress direction of the first animal and the second direction. All three conditions are required: the condition of the angle to the position of the animal (angle condition), and the condition of whether the first animal is approaching and the second animal is leaving (approaching condition). When the conditions are established at the same time, it can be determined that the first animal is tracking the second animal. And, as an evaluation index of the tracking behavior,

Tracking time rate-tracking time Observation time

Tracking distance ratio = Moving distance while tracking Z Total moving distance

Etc. can be used.

Further, the animal behavior analysis method of the present invention includes: an observation data acquisition step of acquiring a movement locus of an animal; and a fractal dimension calculation step of calculating a fractal dimension of the movement locus acquired in the observation data acquisition step. It is characterized by including.

Further, the animal behavior analysis system of the present invention comprises: observation data acquisition means for acquiring a movement locus of an animal; and fractal dimension calculation means for calculating a fractal dimension of the movement locus acquired by the observation data acquisition means. It is specially equipped.

The above method and configuration quantitatively evaluate the complexity of the animal's trajectory. Worth it. The animal may be an aquatic animal such as zebrafish or a terrestrial animal such as drosophila and mouse.

Further, the animal behavior analysis program of the present invention is a computer program that causes a computer to function as each of the above means.

With the configuration of the upper IE, by implementing each means of the upper IH animal behavior analysis system using a computer, the above animal behavior analysis system can be realized. Therefore, as the animal behavior analysis system described above, it becomes possible to analyze the behavior of the animal precisely and regularly, to the extent that it can respond to the needs of gene research.

Further, a computer-readable recording medium on which the animal behavior analysis program of the present invention is recorded is a computer-readable recording medium that realizes each of the above-mentioned IS and operates the above-described animal behavior analysis system. This is a computer-readable recording medium recorded by IB.

With the configuration described above, the behavior analysis system for animals can be realized on a computer by using the behavior analysis program for office work read from the recording medium.

Still other objects, features, and strengths of the present invention will be made clear by the description below. Also, the advantages of the present invention will become apparent in the following description with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is an explanatory diagram schematically showing a configuration of a behavior analysis system according to one embodiment of the present invention.

Fig. 2 shows the decision made by the tracking behavior evaluation unit of the behavior analysis system shown in Fig. 1. It is a figure explaining the distance condition between individuals. .

Figs. 3 (a) and 3 (b) are diagrams illustrating the angle condition determined by the tracking behavior evaluation unit of the behavior analysis system shown in Fig. 1.Fig. 3 (a) shows the case where the angle condition is satisfied. 3 (b) shows the case where the angle condition is satisfied but the approach condition is not satisfied.

Figs. 4 (a) and 4 (b) are diagrams illustrating the approach conditions determined by the tracking behavior evaluation unit of the behavior analysis system shown in Fig. 1.Fig. 4 (a) is a diagram when the approach conditions are satisfied. 4 (b) shows the case where the approach condition is not satisfied. Fig. 5 shows an example of the analysis of the tracking behavior of Zebrafish, using the tracking time rate calculated by the tracking behavior evaluation unit of the behavior analysis system shown in Fig. 1 to quantitatively evaluate the effects of optic nerve transection. It is a graph.

FIG. 6 is a graph showing the results of observing the recovery of the visual function of zebrafish using the tracking time rate and the tracking distance rate calculated by the tracking action evaluation unit of the action analysis system shown in FIG.

FIG. 7 is a diagram illustrating a method of calculating a fractal dimension of a fish swimming locus in a fractal dimension calculating unit of the behavior analysis system shown in FIG. FIG. 8 is a diagram for explaining a method of calculating a fractal dimension of a swimming locus of a fish in a fractal dimension calculating unit of the behavior analysis system shown in FIG. Figure 9 shows an example of analyzing the swimming trajectory of a zebrafish, using the fractal dimension calculated by the fractal dimension calculator of the behavior analysis system shown in Fig. 1 to quantitatively evaluate the effect of optic nerve transection. Figure 10 shows the swimming trajectory of the zebrafin fry immediately after hatching (within 3 days) with a body length of less than 1 cm, taken by the behavior analysis system shown in Fig. 1. (30 minutes).

Fig. 11 is a photograph showing the swimming locus of zebrafish (30 minutes) one month after hatching with a body length of 2 cm and taken by the behavior analysis system shown in Fig. 1.

Fig. 12 is a drawing substitute photograph showing the swimming locus (30 minutes) of adult zebrafish fish four months after hatching with a body length of 4 cm and hatched by the behavior analysis system shown in Fig. 1.

Fig. 13 is a drawing substitute photograph showing the swimming trajectory (10 minutes) of two normal zebrafish, taken by the behavior analysis system shown in Fig. 1.

Fig. 14 is a drawing substitute photograph showing the swimming locus (10 minutes) of two blind zebrafish taken by the behavior analysis system shown in Fig. 1. BEST MODE FOR CARRYING OUT THE INVENTION

One embodiment of the present invention will be described below with reference to FIGS. In this embodiment, as an example, a description will be given of a case in which the behavioral analysis of zebrafish, in particular, the case of quantitatively evaluating the effect of optic nerve transection (visual blockage).

As shown in FIG. 1, the behavior analysis system 1 according to the present embodiment includes a water tank 2 containing zebrafish F in water, two CCD cameras 3.3, two monitors 4.4, and a video mixer 5 And an image processing device 10. The behavior analysis system 1 performs behavior analysis of the zebrafish F based on a surface image of the zebrafish F in the water tank 2. Aquarium 2 is the observation space. The shape of the water tank 2 is, for example, 24 cm long, 37 cm wide, and 29 cm high. In the aquarium 2, one or more zebrafish F are swimming freely. The aquarium 2 is illuminated by four lamps (.150 W each) (not shown) installed diagonally above the aquarium 2 to prevent reflection and backlight from occurring on the CCD camera 3.

The CCD cameras 3 and 3 are installed above and about 1.5 m from the side of the aquarium 2, and are set so that the entire aquarium 2 can be photographed in two directions: upward and sideways. In addition, the CCD camera 3.3 has a zebrafish F with a small size and a high swimming speed of 70 mm / s, so that the shutter speed is 1 to 60 seconds and the frame rate is 10 sheets. Seconds, Resolution (pixels) You can shoot with settings such as 25 6 (vertical) X 1 28 (horizontal) or 25 6 (vertical) X 25 6 (horizontal). The color tone is full color and a composite video terminal is used for the video signal transmission connector.

Thus, when the experimental animal is zebrafish, the CCD camera

The frame rate 3 is preferably S sheets / sec or more, more preferably 10 sheets / sec. In the case of goldfish, 4 fish / sec was sufficient.

The monitor 4 is a display monitor having a composite video input terminal for displaying an image from the CCD camera 3. The monitor 4 allows the user to check the image being shot with the CCD camera 3 in real time.

The video mixer 5 combines the two images captured by the two CCD cameras 3 and 3 into a single upper and lower image, and outputs it to an image input board 11 (described later).

The image processing device 10 captures an image captured by the CCD camera 3 and 3 010979

Ten

The data is binarized in real time and stored in the data storage unit 13. After the observation, analysis is performed based on the binarized image. For this purpose, the image processing device 10 includes an image input board 11, a binarization processing unit 12, a data storage unit 13, and an analysis processing unit 1.

It has four.

The image input board 11 is an interface of the image processing device 10.

, It is a full-color one-image input board. In the present embodiment, the effective range of the image is set to 512 × 512, and is used in the frame mode. The I-image input board 11 can output an input image from a composite video terminal (snow 1 ^ 1 out) to an external monitor (not shown).

The binarization processing unit 12 compares the image captured from the image input board 11 with the threshold value for each pixel and records the binarized image in the data storage 13 I do.

The data storage unit 13 is a storage device such as a hard disk, and stores a binarized image generated by the binarization processing unit 12 and a calculation result by the analysis processing unit 14.

The analysis processing unit 14 recognizes the zebrafish F based on the binarized image stored in the data storage 13 and performs general measurement of a swimming locus and the like, a tracking action evaluation and a swimming locus. Performs special measurement for total dimension calculation. Therefore, the analysis processing unit 14 includes a general measurement unit 14A, a tracking behavior evaluation unit 14B, and a fractal dimension calculation unit 14C. The analysis results of the general measurement and the special measurement can be stored in the data storage unit 13 or output to a monitor or a printer (not shown).

The general measurement unit 14A converts the observation data (position, speed, swimming trajectory, etc.) of Zepprait F based on the binarized image generated by the binarization processing unit 12. decide. Specifically, the general measurement unit 14A first reads the binarized W image from the data storage 13 and recognizes zebrafish F from the difference in color from the background. Next, the coordinates of the center of gravity of the region of Zebrahui Shish F are obtained. Then, based on the data of the center of gravity, the position coordinates, swimming locus (two-dimensional, three-dimensional), position distribution, moving speed, moving distance, body axis tilt, left and right rotation (right rotation, left rotation) Rotation, straight ahead, etc.). The general measurement unit 14 部 performs image processing such as labeling processing and separation processing in order to prevent rapid separation of surface images and apparent overlap. This makes it possible to handle multiple fish.

Furthermore, when shooting fish that swim at high speed and complexity, such as zebrafish, to increase the frame rate, the binarization is first performed at a quarter resolution, for example, Only the area around the area determined to be present may be binarized at the original resolution. As a result, the number of processes can be significantly reduced, and a favorable frame rate and resolution can be obtained.

The tracking behavior evaluation unit 14B reads the position and velocity, which are the observation data determined by the general measurement unit 14A, from the data storage unit 13 and evaluates the tracking behavior of the target individual to other individuals. I do.

The fractal dimension calculating unit 14C reads the swimming trajectory, which is the observation data determined by the general measuring unit 14A, from the data storage unit 13 and calculates the fractal dimension of the swimming trajectory.

The tracking behavior evaluation unit 14B and the fractal dimension calculation unit 14C will be described later in detail.

Based on the above, the behavior analysis system 1 first allowed the zebrafish F to swim freely in the aquarium 2, Images taken continuously for a predetermined time by the D camera 3 3 are taken into the image processing device 10, binarized and stored in the data storage unit 13. After observation at a predetermined time, the analysis processing unit 14 recognizes zebrafish F from the binary image and performs various analyzes of general measurement and special measurement.

Here, FIGS. 10 to 12 show the swimming locus of one zebrafish F taken by the behavior analysis system 1 for 30 minutes. Figure 10 shows the swimming trajectory of zebrafish F juveniles that have just hatched (less than 3 S) with a body length of less than 1 cm. Figure 11 shows the swimming trajectory of zebrafish F one month after hatching with a body length of 2 cm. Figure 12 shows the swimming locus of adult zebrafish F four months after hatching with a length of 4 cm.

FIGS. 13 and 14 show the swimming locus of two zebrafish F for 10 minutes taken by the behavior analysis system 1. The red and black lines indicate the swimming locus of one zebrafish F, respectively. Figure 13 shows the swimming trajectory of the normal zebrafish F at the tail. According to FIG. 13, it can be seen that the frequency of two normal zebrafish F swimming on the edge of the aquarium 2 is small. In contrast, Figure 14 shows the swimming tracks of two blind zebrafish F. According to FIG. 14, it can be seen that two blind zebrafish F are swimming around the entire aquarium 2.

As described above, according to the behavior analysis system 1, swimming traces of two or more zebrafish F can be obtained. Then, based on the data of the swimming trajectory, it is possible to quantitatively analyze an interaction such as approaching two fishes.

Subsequently, the tracking action evaluation and the fractal dimension calculation of the swimming trajectory, which are special measurements, will be described in detail. (1) Tracking behavior evaluation

The tracking action evaluation performed by the tracking action evaluation unit 14B will be described with reference to FIGS.

Two or more fish, such as zebrafish, form a school and have the habit of swimming as if the following fish followed the preceding fish. The tracking action evaluation unit 14B determines three conditions, (i) an inter-individual distance condition, (ii) an angle condition, and (iii) an approach condition, respectively, and when all are simultaneously established, the tracking action is determined. It is determined that the condition holds. Then, the tracking action evaluation unit 14B evaluates the tracking action by time and distance as a tracking rate.

(i) Individual listening distance condition

If the distance L between the two tails is too far apart, it is not regarded as tracking. Furthermore, if it is too close when viewed from the top of the aquarium 2, it can be determined that swimming is occurring at a position that is overlapping, that is, at a different depth, so that it is determined that tracking is not performed. In the present embodiment, empirically,

l c m <L <10 cm

Was set. However, as shown in Fig. 2, swimming may occur in parallel, so the following conditions are determined.

(ii) Angle condition

In the tracking state, there should always be a chase fish in front of the chase fish. Therefore, as shown in Fig. 3 (a), the condition is that there is a fish that is being chased within a certain angle 0 from the traveling direction of the chasing fish. In the present embodiment, as an angle condition, empirically, Θ ≤ 50 ^β

Was set. However, when facing each other and swimming, as shown in Fig. 3 (b) Therefore, the following condition is determined.

(i i i) Approach conditions

In the tracking state, the chase fish approaches and the chase fish moves away. Therefore, as shown in Fig. 4 (a), when L, L1, and L2 are determined, the approach condition is

L> L 1 and L <L 2

It becomes. FIG. 4B shows an example when tracking is not determined (L> L1 and L> L2).

Then, when all of the above three conditions are satisfied at the same time, the tracking action evaluation unit 14 B sets the tracking action evaluation index as:

Tracking time rate = tracking time Z observation time

Tracking distance ratio = travel distance while tracking / total travel distance

Is calculated.

Thus, Ri by the tracking time rate and tracking distance rate calculated by the tracking behavior evaluation unit 1 4 B, the visual function recovery, it is possible to objectively evaluate from behavioral analysis of goldfish Ya zebrafish _s

Figure 5 shows the results of an analysis of the tracking behavior of zebrafish 'mesh using the tracking time rate. On the left is the tracking rate of normal zebrafish (control experiment), and on the right is the tracking rate of zebrafin on day 1 of bilateral optic nerve transection. About 30% are normal and about 8% are cut, indicating that tracking becomes extremely difficult when blind.

Also, unlike mammals, fish optic nerves regenerate when cut. Therefore, it is extremely important to evaluate whether the fish have restored visual function. Figure 6 shows the recovery of the visual function of the Zeppraite tissue with the tracking time rate and tracking distance. The result of observation using the separation rate is shown. As shown in Fig. 6, when the optic nerve was cut unilaterally or bilaterally, the behavior of the fish was observed, and it was found that optic nerve regeneration had two phases: fast recovery and slow recovery.

That is, ① When the 1 side is cut, the vertical axis of the fish tilts to the normal optic nerve side and rotates well to the normal optic nerve side. The recovery of tilt and rotation was completed in about one month for goldfish and about two weeks for zebrafish. This recovery was confirmed by analyzing the tilt and rotation direction using the general measurement section 14A.

Also, (2) when the optic nerve is cut bilaterally, the tracking behavior is greatly disturbed. The recovery of this tracking action was completed in 4-5 months for goldfish and 2-3 months for Zeprfish. This recovery was confirmed by analyzing the tracking behavior by the tracking behavior evaluation unit 14B.

As described above, the inventor has confirmed that the behavior analysis system 1 can objectively evaluate visual function recovery from the analysis of fish behavior. The values of the tracking time rate and tracking distance rate for 1 day and 1

8%) has been confirmed to be equal to the tracking time rate and tracking distance rate calculated from an image obtained by superimposing two different images taken with only one zebrafish F in water tank 2. Therefore, about 8% can be ignored as a coincidence that the two zebrafish accidentally have a positional relationship that satisfies the tracking determination condition.

(2) Fractal dimension of swimming trajectory

The evaluation of the complexity of the swimming locus of the fish by the fractal dimension calculating unit 14C will be described with reference to FIGS. In the present embodiment, the fractal dimension is a parameter representing the complexity of the curve. As shown in Fig. 7, assuming that the number of measures required when approximating a curve using a measure of length ε is Ν (Ϊ), the fractal dimension D is given by the following relational expression:

Ν (ε) = k ■ ε ' ^-D

Is defined by Taking the log of both sides of this equation gives

logN (ε) =-D ■ log s + k '

It becomes. Here, k and k 'are constants.

Here, if the curve has fractal properties, N ( _ε ) and

This means that when you plot £ on a logarithmic graph, it is arranged in a straight line over a certain range. Therefore, as shown in FIG. 8, the fractal dimension D can be obtained by obtaining the regression line of the points on the graph using the least squares method and examining the slope.

Figure 9 shows the results of analyzing the swimming trajectory of zebrafish using fractal dimensions. On the left is the fractal dimension of a normal zebrafish (control experiment), and on the right is the fractal dimension of the first zebrafish after bilateral optic nerve transection. The larger the value of the fractal dimension is, the more complicated the trajectory of the swim is. Therefore, the results show that the trajectory becomes simpler when blind.

As described above, the behavior analysis system 1 consists of a CCD camera 3 that captures images of zebrafish F in the aquarium 2 at a frame rate of 8 or more per second, and a general measurement unit 1 that acquires observation data from the images. 4 A. Then, the tracking behavior evaluation unit 14B evaluates the tracking behavior between the zebrafish F based on the position velocity, which is the observation data of the zebrafish F. The fractal dimension calculator 14C calculates the fractal dimension of the swimming trajectory based on the swimming trajectory that is the observation data of zebrafish F. You.

This makes it possible to quantitatively and precisely analyze the behavior of zebrafish, which is optimal as an experimental animal for genetic research, but swims at high speed and in a complicated manner. Then, the complex behavior of the zebrafish can be obtained in real time and quantitatively analyzed. In addition, the behavior of zebrafish can be easily measured from immediately after hatching to adult fish.

Note that the present embodiment does not limit the scope of the present invention, and various changes can be made within the scope of the present invention. For example, the present invention can be configured as follows.

In order to evaluate the tracking behavior, it is sufficient that the position and speed of the animal can be used as observation data, and in order to calculate the fractal dimension, it is sufficient that the movement trajectory can be used as observation data. Therefore, instead of generating these observation data from images, the observation data may be acquired by, for example, position detection using a transmitter mounted on an animal.

In addition, a feeding device is installed in the water channel 2, and this feeding device, CCD camera 3, video mixer 5, image processing device 10 (image input board 11, binarization processing unit 12, data storage unit 1) By controlling 3) with a timer, long-term observations as shown in Fig. 6 can be performed automatically.

According to the behavior analysis system 1, it is possible to standardize the behavior analysis of fish and the like. Therefore, the behavior analysis system 1 can be variously used in a field that uses quantification of animal behavior analysis. For example, as described above, quantitative analysis of abnormal behavior after optic nerve transection (visual blockage) is possible. That is, it can be used for evaluation of nerve damage, regeneration, etc. It can also be used for genetic modification and screening for mutant phenotypic abnormalities. 'Comparison between nocout fish and wild fish allows us to analyze the correlation between gene and phenotype (behavior). Furthermore, it can be used to evaluate the effects and duration of drug effects (especially neuroactive drugs).

In addition, the behavior analysis system 1 can be used for screening external factors (water quality, environmental hormones). Therefore, it is possible to monitor the deep swimming boundaries of fish (dissolved oxygen, environmental sources, water temperature, poisons, etc.).

In addition, the behavior analysis system 1 can be used to compare natural fish with artificially reared fish, and to analyze the behavior of schools of fish. Therefore, the behavior analysis system 1 can be applied to fishing techniques such as fishing and aquaculture.

Furthermore, the behavior analysis system 1 can be applied to three-dimensional behavior observation of terrestrial animals such as Drosophila and mice, as well as aquatic animals such as zebrafish. It can also be applied to the behavioral psychology of animals, for example, by observing changes in fish behavior by applying colors and patterns to the walls of the aquarium.

Finally, the image processing apparatus 10 can be configured based on a general-purpose computer such as a workstation or a personal computer. Ie

The image processing device 10 includes a CPU (central processing unit) that executes instructions of a program for realizing the function, a ROM (read only memory) storing boot logic, and a RAM (random access memory), storage devices (recording media) such as a hard disk for storing the top sc programs and various databases, input devices such as keyboards and mice, and output devices such as moyuta, speakers and printers. Are connected to each other.

Further, an object of the present invention is to provide a program for executing an animal behavior analysis program (executable program, A recording medium in which an intermediate code program and a source program) are recorded so as to be readable by a computer is supplied to a system or an apparatus, and the computer (or CPU, MPU, or DSP) of the system or the apparatus is recorded on the recording medium. This can also be achieved by reading and executing a program code. In this case, the program code itself read from the recording medium realizes the above-described function, and the recording medium on which the program code is recorded constitutes the present invention.

More specifically, the binarization processing unit 12 and the analysis processing unit 14 included in the image processing device execute a predetermined program stored in a memory (not shown) of the surface image processing device 10. This is realized by execution by a microprocessor or the like.

The recording medium for supplying the program code can be configured to be separable from the system or the device. Further, the recording medium may be a medium which is fixedly supported so that a program code can be supplied. The recording medium is connected to the system or the device as an external storage device even if the recording medium is mounted on the system or the device so that the recorded program code can be directly read by the computer. It may be mounted so that it can be read via a connected program reader.

For example, the recording medium may be a tape such as a magnetic tape or a cassette tape, or a magnetic disk such as a floppy (registered trademark) disk hard disk. CD-ROM / MO / MD / DVD / Discs including optical discs such as CD-R, IC cards (including memory cards), force cards such as Z optical cards, and some masks ROM / EP ROM / EEPROM flash. A semiconductor memory system such as a flash ROM can be used.

The program code may be read out from the recording medium and directly executed by the computer, or may be transferred from the recording medium to the program storage area of the main memory, and then transferred to the main memory by the computer. It may be recorded so that it can be read out from and executed.

Further, the system or device may be configured to be connectable to a communication network, and the program code may be supplied via the communication network. The communication network is not particularly limited. Specifically, the Internet, the intranet, the extranet, the LAN, the ISDN, the VAN, the CATV communication network, and the virtual private network ( virtual private network), telephone line network, mobile communication network, satellite communication network, etc. are applicable. Further, the transmission medium constituting the communication network is not particularly limited, and specific examples include IEEE1394, USB, power line carrier, cable TV line, telephone line, and ADSL line. It can be applied to wireless communication such as infrared communication such as IrDA and remote control, Bluetooth, 80.2.11 wireless, HDR, mobile phone network, satellite line, and terrestrial digital network. Note that the present invention can also be realized in the form of a carrier wave or a data signal sequence in which the program code is embodied by electronic transmission.

The program for reading the program code from the recording medium and storing the program code in the main memory and the program for downloading the program code from the communication network can be executed by a computer beforehand in a system or apparatus. It shall be stored.

The functions described above are realized not only by executing the above-described program code read by a computer, but also by executing the program code. Based on the instructions, the OS running on the computer performs the actual processing.

—It is also realized by performing part or all.

Further, in the above-described function, the program code read from the recording medium is written in a memory provided in a function expansion board connected to a function expansion board attached to a computer or a computer. Thereafter, based on the instructions of the program, the CPU or the like provided in the function expansion board or the function expansion unit performs a part or all of the actual processing.

Specific embodiments or examples made in the section of the detailed description of the invention clarify the technical contents of the present invention, and are limited to only such specific examples. However, the present invention can be variously modified and implemented within the spirit of the present invention and the scope of the claims. Industrial potential

The animal behavior analysis method and the animal behavior analysis system according to the present invention, which are optimal as experimental animals for genetic research, can quantitatively and precisely analyze the behavior analysis of zebrafish having a high swimming speed. It allows you to do that. Therefore, the animal behavior analysis method and the animal behavior analysis system according to the present invention can be used for precise quantitative behavior analysis for genetic research. Further, the animal behavior analysis method and the animal behavior analysis system according to the present invention can be widely applied to behavioral analysis of animals that perform high-speed and complicated behaviors, in addition to experimental animals for genetic research.

Claims

The scope of the claims

1) an imaging step of imaging aquatic animals in the aquarium at a frame rate of 8 or more per second;

An analysis step and a knob for analyzing the behavior of aquatic animals based on the image captured in the imaging step.

2. The animal behavior analysis method according to claim 1, wherein the aquatic animal is a zebrafish.

3. An animal behavior analysis system that analyzes the behavior of the aquatic animal based on an image of the aquatic animal in the aquarium,

Up? An animal behavior analysis system equipped with imaging means that captures a B-side image at a frame rate of eight or more Z-seconds.

4. Observation data acquisition step for acquiring the position and velocity of the first animal and the second animal;

A tracking behavior evaluation step of evaluating a tracking behavior of the first animal with respect to the second animal based on the position and velocity of the first animal and the second animal acquired in the observation data acquisition step. Animal behavior analysis method including:

5. Observation data acquisition means for acquiring the position and velocity of the first animal and the second animal;

Tracking behavior evaluation means for evaluating the tracking behavior of the first animal with respect to the second animal based on the positions and velocities of the first animal and the second animal obtained by the observation data obtaining means; Behavior analysis system for animals

6. Observation data acquisition step for acquiring the trajectory of the animal; A fractal dimension calculating step of calculating a fractal dimension of the movement locus acquired in the observation data acquisition step.

7. Observation data acquisition means for acquiring the trajectory of the animal,

A fractal dimension calculating means for calculating a fractal dimension of the movement trajectory acquired by the observation data acquiring means.

8. An animal behavior analysis program for operating the animal behavior analysis system according to claim 5 or 7, which causes a computer to function as each of the above means.

9. A computer-readable recording medium on which the animal behavior analysis program according to claim 8 is recorded.