JP2004089027A - Method for analyzing behavior of animal, system for analyzing behavior of animal, program for analyzing behavior of animal, and recording medium recording the program and readable with computer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To carry out a precise quantitative behavioral analysis so as to meet even requests of genetic studies. <P>SOLUTION: A system 1 for analyzing behaviors is equipped with a charge-coupled device (CCD) camera 3 picking up images of zebrafish F in an aquarium 2 at ≥8 frames/s frame rate and a usual measuring part 14A obtaining observed data from the images. A part 14B for evaluating chasing behaviors evaluates the mutual chasing behaviors of the zebrafishes F based on the positions and speeds which are the observed data of the zebrafishes F. The fractal dimension of swimming loci is computed on the basis of swimming loci which are observed data of the zebrafishes F with a fractal dimension computing part 14C. Thereby, the behavioral analysis of the zebrafish having a high swimming speed though optimal as an experimental animal for the genetic studies can quantitatively and precisely be carried out. <P>COPYRIGHT: (C)2004,JPO

Description

[0001]
TECHNICAL FIELD OF THE INVENTION
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 the behavior of an animal, an animal behavior analysis system, an animal behavior analysis program, and The present invention relates to a recorded computer-readable recording medium.
[0002]
[Prior art]
The development of genetic research in recent years has been remarkable, and the research target has shifted from lower organisms such as nematodes, Drosophila and yeast to higher animals such as mice, fish and humans. In the genetic research, the post-genome after the analysis of the whole genome has entered the era of proteomics, in which the function of the protein encoded by the genome is analyzed, and the era of phenomics, in which the phenotype is analyzed. It is expected to move to F.
[0003]
Here, zebrafish has been widely recognized as the most suitable animal species for genetic modification and gene knockout because of its extremely advantageous features in developmental biology. It has become one of.
[0004]
Specifically, zebrafish is a small tropical fish with a length of 5 to 6 cm. Unlike other fish, (1) eggs are laid at any time of the year, so eggs can be obtained throughout the year. Since the body is transparent, it has the feature that gene expression can be tracked and observed from outside with a fluorescent protein such as GFP (green fluorescent protein).
[0005]
In zebrafish genetic research, analysis of the whole genome is imminent, and in the future, the demand for behavioral analysis of fish as a phenotype, including developmental biology, that is, behavioral analysis of fish that has undergone genetic modification and gene knockout is immeasurable. Absent.
[0006]
However, behavior analyzers for small animals such as rats and mice are often used for medical research and are commercially available, but at present there are few fish analyzers.
[0007]
Note that the present inventors have disclosed Kato, S.A. Tamada, K .; , Shimada, Y .; {And} Chujo, T .; , A quantification of goldfish behavior by an image processing system, Behavioural Brain Research, 80 (1996) 51-55. In this paper, we proposed an image processing system for analyzing the behavior of goldfish, which captures the goldfish in the aquarium and analyzes the position distribution and direction of travel of the goldfish. Using the image processing system, the inventors have analyzed the behavior of goldfish, and in particular, performed a quantitative analysis of abnormal behavior after optic nerve transection.
[0008]
In addition, the published patent publication “Japanese Patent Application Laid-Open No. Hei 7-63747 (published on March 10, 1995)” discloses an abnormal water quality by discriminating the activity status and the survival status of the fish school in the aquarium. "Water quality inspection device" for detecting the water content is described. In this water quality inspection device, an image of a school of fish released into an aquarium is taken by an ITV camera, and a sampling process and a binarization process are executed by an image input unit. By setting the threshold value for binarization, the influence of disturbance or noise can be eliminated. The image processing unit extracts an image of each fish constituting the fish school from the binarized image data, attaches a label to each image, and based on the movement amount per unit time, the activity status and the survival status of the fish school. Perform identification.
[0009]
[Problems to be solved by the invention]
As described above, despite the necessity and urgent need for quantitative analysis of the behavior of zebrafish, there has been no suitable behavior analysis method or behavior analysis system. In other words, in the behavior analysis system and water quality inspection device for goldfish, since zebrafish perform high-speed and complicated three-dimensional behavior, it is not possible to perform precise quantitative behavior analysis to the extent that it can meet the demands of genetic research. It was possible.
[0010]
The present invention has been made to solve the above problems, and an object of the present invention is to provide an animal behavior analysis method and an animal capable of performing a quantitative behavior analysis that is precise enough to meet the requirements of genetic research. An object of the present invention is to provide a behavior analysis system. Another object of the present invention is to provide an animal behavior analysis program for realizing the above-described animal behavior analysis system, and a computer-readable recording medium recording the program.
[0011]
[Means for Solving the Problems]
In order to solve the above problem, the animal behavior analysis method of the present invention is based on an imaging step of imaging an aquatic animal in an aquarium at a frame rate of 8 or more per second, based on the image captured in the imaging step. And analyzing the behavior of the aquatic animal.
[0012]
Furthermore, the animal behavior analysis method of the present invention is characterized in that the aquatic animal is zebrafish.
[0013]
Further, the animal behavior analysis system of the present invention is an animal behavior analysis system for analyzing the behavior of aquatic animals based on an image of an aquatic animal in an aquarium. It is characterized by having an imaging means for imaging at a frame rate.
[0014]
According to the above-described method and configuration, by capturing images serving as a basis for behavior analysis at a frame rate of 8 or more per second, behavior analysis of aquatic animals that perform complex behavior at high speed becomes possible.
[0015]
As explained in the prior art, zebrafish has extremely advantageous features in developmental biology, and phenotypic behavior analysis, that is, quantification of the behavior of zebrafish that has been genetically modified or knocked out There is a strong demand for the establishment of a technique for performing analytical analysis.
[0016]
Therefore, according to the above method and configuration, even for aquatic animals such as zebrafish that perform high-speed and complicated three-dimensional behavior, the behavior can be reliably imaged, so that behavior analysis becomes possible. . Therefore, it becomes possible to analyze the behavior of zebrafish precisely and quantitatively to the extent that it can meet the demands of genetic research.
[0017]
Further, the animal behavior analysis method of the present invention includes an observation data acquiring 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 acquiring step. 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 the velocity of the animal.
[0018]
Also, the animal behavior analysis system of the present invention includes an observation data acquisition unit that acquires the positions and velocities of the first animal and the second animal, and the first animal and the second animal that are acquired by the observation data acquisition unit. Tracking behavior evaluation means for evaluating the tracking behavior of the first animal with respect to the second animal based on the position and velocity of the animal.
[0019]
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 of different species. The first animal and the second animal may be aquatic animals such as zebrafish or terrestrial animals such as Drosophila and mice.
[0020]
Here, as a specific evaluation method of the tracking behavior, for example, the condition of the distance between the first animal and the second animal (distance between individuals), the traveling direction of the first animal, and the When all three conditions of the angle with the position (angle condition) and the condition of whether the first animal is approaching and the second animal is moving away (approaching condition) are satisfied at the same time, the first condition is satisfied. Can be determined to be following 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 / total moving distance
Etc. can be used.
[0021]
Further, the animal behavior analysis method of the present invention includes an observation data acquisition step of acquiring a movement trajectory of the animal, and a fractal dimension calculation step of calculating a fractal dimension of the movement trajectory acquired in the observation data acquisition step. It is characterized by.
[0022]
Further, the animal behavior analysis system of the present invention includes observation data acquisition means for acquiring a movement trajectory of the animal, and fractal dimension calculation means for calculating a fractal dimension of the movement trajectory acquired by the observation data acquisition means. It is characterized by doing.
[0023]
According to the above method and configuration, the complexity of the movement trajectory of the animal can be quantitatively evaluated. The animals may be aquatic animals such as zebrafish or terrestrial animals such as Drosophila and mice.
[0024]
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.
[0025]
With the above configuration, the computer can realize each unit of the animal behavior analysis system, thereby realizing the animal behavior analysis system. Therefore, as the above-described animal behavior analysis system, it becomes possible to analyze the behavior of the animal precisely and quantitatively to such an extent that it can meet the demands of genetic research.
[0026]
In addition, a computer-readable recording medium on which the animal behavior analysis program of the present invention is recorded is a computer that stores an animal behavior analysis program for operating the above-described animal behavior analysis system by causing the computer to realize the above-described units. This is a readable recording medium.
[0027]
With the above configuration, the animal behavior analysis system can be realized on a computer by the animal behavior analysis program read from the recording medium.
[0028]
BEST MODE FOR CARRYING OUT THE INVENTION
One embodiment of the present invention will be described below with reference to FIGS. In the present embodiment, as an example, a case will be described in which the behavior analysis of zebrafish, in particular, the case of quantitatively evaluating the effect of optic nerve transection (visual blockage) is performed.
[0029]
As shown in FIG. 1, a 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, a video mixer 5, An image processing apparatus 10 is provided. The behavior analysis system 1 analyzes the behavior of the zebrafish F based on an image of the zebrafish F in the water tank 2.
[0030]
The water tank 2 is an observation space. The shape of the water tank 2 is, for example, 24 cm long, 37 cm wide, and 29 cm high. One or a plurality of zebrafish F are swimming freely in the water tank 2. The water tank 2 is illuminated by four reflex lamps (150 W each) (not shown) installed diagonally above the water tank 2 so as to prevent reflection and backlight from occurring on the CCD camera 3.
[0031]
The CCD cameras 3, 3 are installed above the water tank 2 and at a position of about 1.5 m laterally, and are set so that the entire water tank 2 can be photographed in two directions, that is, upward and laterally. In addition, since the zebrafish F is small and the swimming speed is as fast as 70 mm / sec, the CCD camera 3.3 has a shutter speed of 1/60 sec, a frame rate of 10 frames / sec, and a resolution (pixel) in the upward direction. Images can be taken at 256 (vertical) x 128 (horizontal) in the horizontal direction. The photographing is performed, for example, continuously for 30 minutes. The color tone is full color. Also, a composite video terminal is used for a video signal transmission connector.
[0032]
As described above, when the experimental animal is zebrafish, the frame rate of the CCD camera 3 is preferably 8 frames / second or more, and more preferably 10 frames / second. In the case of goldfish, 4 frames / sec was sufficient.
[0033]
The monitor 4 is a display monitor having a composite video input terminal for displaying an image from the CCD camera 3. With the monitor 4, an image being captured by the CCD camera 3 can be confirmed in real time.
[0034]
The video mixer 5 combines the two images captured by the two CCD cameras 3 into one image divided into upper and lower parts, and outputs it to an image input board 11 (described later).
[0035]
The image processing device 10 captures an image captured by the CCD camera 3, binarizes the image in real time, records it in the data storage unit 13, and performs analysis based on the binarized image after the observation. To this end, the image processing apparatus 10 includes an image input board 11, a binarization processing unit 12, a data storage unit 13, and an analysis processing unit 14.
[0036]
The image input board 11 is an interface of the image processing device 10 and is a full-color image input board. In the present embodiment, the effective range of the image is 512 × 512 pixels, and the image is used in the frame mode. The image input board 11 can output an input image from a composite video terminal (through-out) to an external monitor (not shown).
[0037]
The binarization processing unit 12 records the binarized image in the data storage unit 13 by comparing the image captured from the image input board 11 with a threshold value for each pixel.
[0038]
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.
[0039]
The analysis processing unit 14 recognizes the zebrafish F based on the binarized image stored in the data storage unit 13 and performs general measurement of a swimming trajectory and the like, and special measurement of tracking action evaluation and fractal dimension calculation of the swimming trajectory. I do. 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).
[0040]
The general measurement unit 14A determines observation data (position, speed, swimming locus, etc.) of the zebrafish F based on the binarized image generated by the binarization processing unit 12. Specifically, first, the general measurement unit 14A reads the binarized image from the data storage unit 13, and recognizes the zebrafish F from the difference in color from the background. Next, the coordinates of the center of gravity of the zebrafish F area are obtained. Subsequently, based on the center-of-gravity coordinate data, position coordinates, swimming trajectory (two-dimensional, three-dimensional), position distribution, moving speed, moving distance, body axis inclination, left and right rotation (right rotation, left rotation, straight ahead, etc. ) And so on. Note that the general measurement unit 14A performs image processing such as labeling processing and occlusion separation processing in order to prevent rapid separation of images and apparent overlap. This makes it possible to handle a plurality of fish.
[0041]
Furthermore, when photographing a fish that swims at high speed and complexity, such as a zebrafish, binarization is first performed at a resolution of, for example, 1/4 to increase the frame rate and resolution, and the fish is determined to be a fish. Only the area around the area may be binarized at the original resolution. As a result, the number of processes can be significantly reduced, and a preferable frame rate and resolution can be obtained.
[0042]
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 focused individual to other individuals.
[0043]
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.
[0044]
As described above, the behavior analysis system 1 first allows the zebrafish F to freely swim in the water tank 2 and continuously captures images of the zebrafish F from the upper and side directions of the water tank 2 with the CCD cameras 3.3 for a predetermined time. Is taken into the image processing apparatus 10, binarized, and stored in the data storage unit 13. After observation for a predetermined time, the analysis processing unit 14 recognizes the zebrafish F from the binarized image and performs various analyzes of general measurement and special measurement.
[0045]
Subsequently, the tracking action evaluation and the fractal dimension calculation of the swimming trajectory, which are special measurements, will be described in detail.
[0046]
(1) Tracking behavior evaluation
The tracking action evaluation by the tracking action evaluation unit 14B will be described with reference to FIGS.
[0047]
Two or more fish, such as zebrafish, form a school and have a 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 a tracking action is established when all of them are established simultaneously. It is determined that there is. Then, the tracking action evaluation unit 14B evaluates the tracking action by time and distance as a tracking rate.
[0048]
(I) Distance condition between individuals
If the distance L between the two animals is too far, it is not regarded as tracking. Further, if the water tank 2 is too close when viewed from above, it can be determined that they are swimming, that is, they are swimming at positions of different depths, so that it is determined that tracking is not performed. In the present embodiment, empirically,
1cm <L <10cm
Was set. However, as shown in FIG. 2, there are times when swimming occurs in parallel.
[0049]
(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. 3A, the condition is that there is a fish being chased within a certain angle θ from the traveling direction of the chasing fish. In the present embodiment, as the angle condition, empirically,
θ = 50 °
Was set. However, as shown in FIG. 3B, there is a case where the swimmers face each other, and therefore the following condition is determined.
[0050]
(Iii) Approach conditions
In the tracking state, the chase fish tries to approach and the chase fish tries to move away. Therefore, when L, L1, and L2 are determined as shown in FIG.
L> L1 and L <L2
It becomes. FIG. 4B shows an example when tracking is not determined (L> L1 and L> L2).
[0051]
When all of the above three conditions are satisfied at the same time, the tracking action evaluation unit 14B sets the tracking action evaluation index as
Tracking time rate = tracking time / observation time
Tracking distance ratio = moving distance while tracking / total moving distance
Is calculated.
[0052]
Therefore, based on the tracking time rate and the tracking distance rate calculated by the tracking action evaluation unit 14B, it is possible to objectively evaluate visual function recovery from a goldfish or zebrafish action analysis.
[0053]
FIG. 5 shows the result of analyzing the tracking behavior of the zebrafish using the tracking time rate. The left side shows the tracking rate of normal zebrafish (control experiment), and the right side shows the tracking rate of zebrafish on the first day of bilateral optic nerve transection. The normal one is about 30% and the cut one is about 8%, which indicates that tracking becomes extremely difficult when blind.
[0054]
Unlike mammals, fish optic nerves regenerate even when cut. Therefore, it is extremely important to evaluate whether the fish have really restored visual function.
[0055]
FIG. 6 shows the results of observing the recovery of the visual function of zebrafish using the tracking time rate and the tracking distance rate. As shown in FIG. 6, as a result of observing the behavior of the fish when the optic nerve is severed unilaterally or bilaterally, it can be seen that the optic nerve has two phases of fast recovery and slow recovery.
[0056]
That is, when the (1) 1 side is cut, the vertical axis of the fish tilts toward the normal optic nerve side and rotates well toward the normal optic nerve side. This 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 inclination and the rotation direction by the general measuring unit 14A.
[0057]
(2) When the optic nerve is bilaterally cut, the tracking behavior is greatly disturbed. Recovery of this tracking behavior was completed in 4-5 months for goldfish and 2-3 months for zebrafish. This recovery was confirmed by analyzing the tracking action by the tracking action evaluation unit 14B.
[0058]
As described above, the inventor has confirmed that the behavior analysis system 1 can objectively evaluate visual function recovery from fish behavior analysis.
[0059]
The values of the tracking time rate and the tracking distance rate for one day and one week in FIG. 6 (about 8%) are obtained by superimposing two different images obtained by putting only one zebrafish F in the water tank 2. It is confirmed that they are equal to the tracking time rate and the tracking distance rate calculated from. Therefore, about 8% can be disregarded as a coincidence that the two zebrafish accidentally have a positional relationship that satisfies the tracking determination condition.
[0060]
(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.
[0061]
As shown in FIG. 7, assuming that the number of measures required when approximating a curve using a measure of length ε is N (ε), a fractal dimension D is represented by the following relational expression:
N (ε) = k · ε^1-D
Is defined by Taking the log of both sides of this equation gives
logN (ε) = − D · logε + k ′
It becomes. Here, k and k 'are constants.
[0062]
Here, the above equation means that, when the curve has fractal properties, when N (ε) and ε are plotted on a logarithmic graph, they are arranged in a straight line within a certain range. Therefore, as shown in FIG. 8, a fractal dimension D can be obtained by obtaining a regression line of a point on the graph using the least squares method and examining the slope.
[0063]
FIG. 9 shows the result of analyzing the swimming locus of zebrafish using the fractal dimension. The left side shows the fractal dimension of normal zebrafish (control experiment), and the right side shows the fractal dimension of zebrafish on day 1 of bilateral optic nerve transection. The larger the value of the fractal dimension is, the more complicated the trajectory of the swim is. Therefore, it can be understood from the result that the trajectory of the swim becomes simpler when the blindness is increased.
[0064]
As described above, the behavior analysis system 1 includes the CCD camera 3 that captures images of the zebrafish F in the water tank 2 at a frame rate of 8 sheets / second or more, and the general measurement unit 14A that acquires observation data from the images. Prepare. Then, the tracking behavior evaluation unit 14B evaluates the tracking behavior between the zebrafish F based on the position and the speed, which are the observation data of the zebrafish F. Further, the fractal dimension calculating unit 14C calculates the fractal dimension of the swimming trajectory based on the swimming trajectory which is the observation data of the zebrafish F.
[0065]
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 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.
[0066]
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 embodiment can be configured as follows.
[0067]
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, these observation data may be acquired by, for example, position detection using a transmitter mounted on an animal, instead of generating the observation data from an image.
[0068]
Further, a feeding device is provided in the water tank 2, and the feeding device, the CCD camera 3, the video mixer 5, and the image processing device 10 (the image input board 11, the binarization processing unit 12, and the data storage unit 13) are controlled by a timer. Thus, a long-term observation as shown in FIG. 6 can be automatically performed.
[0069]
According to the behavior analysis system 1, quantification of behavior analysis of fish and the like can be realized. Therefore, the behavior analysis system 1 can be variously used in a field that uses quantification of behavior analysis of an animal. 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 and regeneration. In addition, it can be used for screening for genetic modification and mutant phenotypic abnormalities. Further, by comparing a genetically modified fish or a knockout fish with a wild fish, the correlation between the gene and the phenotype (behavior) can be analyzed. Furthermore, it can be used to evaluate the effect of drug efficacy (especially neuroactive drugs and the like) and duration.
[0070]
In addition, the behavior analysis system 1 can be used for screening for external factors (water quality, environmental hormones). Therefore, it is possible to monitor the swimming environment (dissolved oxygen, environmental mutagens, water temperature, poisons, etc.) of the fish.
[0071]
In addition, the behavior analysis system 1 can be used for comparison between natural fish and artificially bred fish and for analysis of fish school behavior, so that it can be applied to fishing techniques such as fishing methods and aquaculture methods.
[0072]
Further, the behavior analysis system 1 can be applied to three-dimensional behavior observation of not only aquatic animals such as zebrafish but also terrestrial animals such as Drosophila and mice. In addition, the present invention can be applied to behavioral psychology of animals, for example, by observing changes in the behavior of fish by adding colors and patterns to the wall surface of the aquarium.
[0073]
Finally, the image processing apparatus 10 can be configured based on a general-purpose computer such as a workstation or a personal computer. That is, the image processing apparatus 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) that expands the program. A storage device (recording medium) such as a hard disk for storing the programs and various databases, input devices such as a keyboard and a mouse, and output devices such as a monitor, a speaker, and a printer are connected by an internal bus.
[0074]
Further, an object of the present invention is to provide a computer-readable recording medium storing program codes (executable format program, intermediate code program, source program) of an animal behavior analysis program which is software for realizing the above-described functions. The present invention can also be achieved by supplying the program to a system or an apparatus and causing a computer (or CPU, MPU, or DSP) of the system or the apparatus to read and execute a program code recorded on a recording medium. 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.
[0075]
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 image processing device 10 by a microprocessor or the like. This is achieved by:
[0076]
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 that fixedly carries the program code so that the program code can be supplied. Even if the recording medium is mounted on a system or an apparatus so that a computer can directly read the recorded program code, the recording medium may be connected to a system or apparatus as an external storage device via a program reading apparatus connected to the system or the apparatus. It may be mounted so that it can be read.
[0077]
For example, the recording medium may be a tape system such as a magnetic tape or a cassette tape, a magnetic disk such as a floppy (registered trademark) disk / hard disk, or a disk including an optical disk such as a CD-ROM / MO / MD / DVD / CD-R. System, a card system such as an IC card (including a memory card) / optical card, or a semiconductor memory system such as a mask ROM / EPROM / EEPROM / flash ROM.
[0078]
Further, the program code may be recorded so that the computer can read out from the recording medium and directly execute the program code, or can be read out from the main storage and executed by the computer after being transferred from the recording medium to the program storage area of the main storage. May be recorded as follows.
[0079]
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, intranet, extranet, LAN, ISDN, VAN, CATV communication network, virtual private network (virtual private network), telephone line network, mobile communication A network, a satellite communication network, or the like is applicable. The transmission medium constituting the communication network is not particularly limited. Specifically, even if the transmission medium is a wire such as IEEE 1394, USB, power line carrier, cable TV line, telephone line, or ADSL line, an infrared ray such as IrDA or remote control is used. , Bluetooth, 802.11 wireless, HDR, mobile phone network, satellite line, digital terrestrial network, and the like.
[0080]
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 are stored in advance in a system or an apparatus so as to be executable by a computer. I do.
[0081]
The functions described above are realized not only by executing the above-described program code read by the computer, but also based on the instruction of the program code, the OS or the like running on the computer performs part or all of the actual processing. It is also realized by performing.
[0082]
Further, the above-described functions are realized by writing the program code read from the recording medium into a memory provided in a function expansion board or a function expansion unit connected to the computer. Is also realized by a CPU or the like provided in the function expansion board or function expansion unit performing part or all of the actual processing based on the instruction.
[0083]
【The invention's effect】
As described above, the animal behavior analysis method of the present invention includes an imaging step of imaging an aquatic animal in an aquarium at a frame rate of 8 sheets / second or more, and a behavior of the aquatic animal based on the image captured in the imaging step. And performing an analysis.
[0084]
Further, in the animal behavior analysis method of the present invention, the aquatic animal is a zebrafish.
[0085]
Further, the animal behavior analysis system of the present invention is an animal behavior analysis system for analyzing the behavior of aquatic animals based on an image of an aquatic animal in an aquarium. This is a configuration including an imaging unit that performs imaging at a frame rate.
[0086]
Therefore, even for aquatic animals such as zebrafish that perform complex three-dimensional behavior at high speed, the behavior can be reliably imaged, and the behavior analysis can be performed. Therefore, there is an effect that the behavior of the zebrafish can be analyzed quantitatively and precisely so as to be able to meet the demands of genetic research.
[0087]
Further, the animal behavior analysis method of the present invention includes an observation data acquiring 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 acquiring step. 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 the velocity of the animal.
[0088]
Also, the animal behavior analysis system of the present invention includes an observation data acquisition unit that acquires the positions and velocities of the first animal and the second animal, and the first animal and the second animal that are acquired by the observation data acquisition unit. Tracking behavior evaluation means for evaluating the tracking behavior of the first animal with respect to the second animal based on the position and velocity of the animal.
[0089]
Therefore, there is an effect that the tracking behavior of the animal can be quantitatively evaluated based on the position and the speed.
[0090]
Also, the animal behavior analysis method of the present invention is a method comprising: 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.
[0091]
Further, the animal behavior analysis system of the present invention includes observation data acquisition means for acquiring a movement trajectory of the animal, and fractal dimension calculation means for calculating a fractal dimension of the movement trajectory acquired by the observation data acquisition means. Configuration.
[0092]
Therefore, there is an effect that the complexity of the moving locus of the animal can be quantitatively evaluated.
[0093]
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.
[0094]
Therefore, the above-described animal behavior analysis system is realized, and the behavior of the animal can be analyzed quantitatively and precisely so as to be able to meet the demands of genetic research.
[0095]
In addition, a computer-readable recording medium on which the animal behavior analysis program of the present invention is recorded is a computer that stores an animal behavior analysis program for operating the above-described animal behavior analysis system by causing the computer to realize the above-described units. This is a readable recording medium.
[0096]
Therefore, there is an effect that the animal behavior analysis system can be realized on a computer by the animal behavior analysis program read from the recording medium.
[Brief description of the drawings]
FIG. 1 is an explanatory diagram showing a schematic configuration of a behavior analysis system according to an embodiment of the present invention.
FIG. 2 is a diagram illustrating an inter-individual distance condition determined by a tracking behavior evaluation unit of the behavior analysis system shown in FIG. 1;
3A and 3B are diagrams illustrating an angle condition determined by a tracking behavior evaluation unit of the behavior analysis system shown in FIG. 1; FIG. 3A illustrates a case where the angle condition is satisfied; FIG. Are not satisfied.
4A and 4B are diagrams illustrating an approach condition determined by a tracking action evaluation unit of the action analysis system shown in FIG. 1, wherein FIG. 4A illustrates a case where the approach condition is satisfied, and FIG. 4B illustrates a case where the approach condition is not satisfied. Shown respectively.
FIG. 5 is a graph showing an example of analyzing the tracking behavior of zebrafish in order to quantitatively evaluate the effect of optic nerve transection using the tracking time rate calculated by the tracking behavior evaluation unit of the behavior analysis system shown in FIG. It is.
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 behavior evaluation unit of the behavior analysis system shown in FIG. 1.
7 is a diagram illustrating a method for 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. 1;
FIG. 8 is a diagram illustrating 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. 1;
9 is a graph showing an example of analyzing a swimming locus of zebrafish in order to quantitatively evaluate the effect of optic nerve transection using the fractal dimension calculated by the fractal dimension calculator of the behavior analysis system shown in FIG. is there.
[Explanation of symbols]
1) Behavior analysis system
2 tank
3 CCD camera (imaging means)
12 binarization processing unit (observation data acquisition means)
14A General measurement section (observation data acquisition means)
14B Tracking behavior evaluation unit (Tracking behavior evaluation means)
14C fractal dimension calculator (fractal dimension calculator)
F zebrafish (aquatic animals)

Claims (9)

An imaging step of imaging aquatic animals in the aquarium at a frame rate of 8 or more per second;
An analyzing step of analyzing the behavior of the aquatic animal based on the image captured in the imaging step. The animal behavior analysis method according to claim 1, wherein the aquatic animal is zebrafish. An animal behavior analysis system for analyzing the behavior of the aquatic animal based on an image of the aquatic animal in the aquarium,
An animal behavior analysis system comprising an image pickup means for picking up the images at a frame rate of 8 sheets / second or more. An observation data acquisition step of acquiring the positions and velocities 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 positions and velocities of the first animal and the second animal acquired in the observation data acquiring step. Characteristic animal behavior analysis method. Observation data acquisition means for acquiring the positions and velocities of the first and second animals, and the first and second animals based on the position and velocity of the first and second animals acquired by the observation data acquisition means. And a tracking behavior evaluation means for evaluating a tracking behavior of the second animal with respect to the second animal. An observation data acquisition step for acquiring an animal movement trajectory;
A fractal dimension calculating step of calculating a fractal dimension of the moving trajectory acquired in the observation data acquiring step. Observation data acquisition means for acquiring a movement locus of an animal;
An animal behavior analysis system, comprising: a fractal dimension calculating unit that calculates a fractal dimension of a movement locus acquired by the observation data acquiring unit. 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. A computer-readable recording medium recording the animal behavior analysis program according to claim 8.

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