RU2437282C2 - Method to assess and preserve morphometric characteristics of biological species in aquaculture - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: invention is designed for use in the sphere of research, monitoring and systematisation of biological species, in particular, when involved into a selection breeding process with aquaculture species. The method consists in picking, fixation, measurements and preservation of aquaculture species parameters. Momental digital photographs are made on the species in macrophotography mode at the researched stage of development. Resolution of digital equipment is at least 300 dpi. Photographs are taken at optimal light at the right angle to the object of research on the background of a metering scale. Measurements are done using a "Ruler" tool in the graphical program "Photoshop" with accuracy of at least 0.1 mm. Linear dimensions of the object are determined by calculation. Computer base is created with characteristics of aquaculture species appearance and morphotype. The produced data is saved in the computer data bank as JPEG or TIFF.

EFFECT: higher reliability of final result of a morphometric description, preservation of the biological species appearance for a long time for further research.

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Description

The invention is intended for use in the field of research, monitoring and systematics of biological objects, in particular when conducting breeding and breeding work with aquaculture objects.

Known are traditional methods for assessing the morphometric characteristics of biological objects, which include collecting material, conducting measurements using instruments with a measurement scale and saving for further analytical work by fixing or preserving in various environments (Pravdin I.F. Guide to the study of fish. - M. : Food industry, 1966. - 380 p.).

In ichthyological studies of the studied relatively large fish, they are subjected to several, usually numerous, measurements with a caliper or a special scoreboard and ruler are used. Linear values are expressed in whole millimeters; small (for example, eye diameter) - with an accuracy of 0.5 mm. Greater accuracy is achieved by the use of Vernier. A micrometer eyepiece is used to measure small objects (caviar or fish larvae).

In population phenetics, in the study of color and spots in salmon fish in the field, various parts of the body of the fish are sketched. The fins are cut off, dried, and later drawn up in the laboratory (Makoedov A.N., Agapova G.A. Methodology of population-phenetic studies of pink salmon according to the variants of the drawing on the tail fin // Marine Biology. - 1991. - No. 5. - P. 92 -94; Makoyedov AN Karyology, biochemical genetics and population phenetics of salmon fish of Siberia and the Far East. - M.: UMK "Psychology", 1999. - P.291).

Carrying out manipulations related to the measurement of living aquatic biological objects intended for further cultivation is associated with the risk of damage and may subsequently be accompanied by pathological changes determined by stress. The effectiveness of obtaining reliable data on measurements in this case is always due to the human factor. The data obtained by different operators on the same attribute of the same individual may differ from each other. The discrepancy between the estimates of different operators measuring the characteristics of one and the same individual can reach a value commensurate with the size of the differences, in ichthyological works usually interpreted as interpopulation (Mina M.V., Levin B.A., Mironovsky A.N. the use in character morphometric studies of fish of character ratings obtained by different operators // Issues of ichthyology. - 2005. - T.45, 3. - S.331-341).

The origin of errors in measuring the length and other parameters of fish larvae is also due to the fact that the fixation, preservation, and preservation of biological material in currently existing media does not completely preserve the external structure, morphotype, and color during long-term storage of the studied objects. The conservation of biological objects in fixing media prevents their objective comparability with living material.

The closest way to the claimed invention in terms of features is a method for assessing the morphometric characteristics of aquaculture objects, which consists in selecting aquaculture objects, fixing them, conducting measurements and saving parameters for further analysis (Krylova V.D., Sokolov L.I. Morphological studies of sturgeon fish and their hybrids. Methodical recommendations. - M.: VNIRO, 1981. - 28 s).

In the known method, the fixation of aquaculture objects is carried out by preservation in special fixing media containing anesthetic or preservative substances. Conducting measurements is also carried out by traditional methods using measuring instruments. Morphometric parameters for further analysis are transferred to special measurement tables.

The reasons that impede the achievement of the technical result indicated below include the complexity and great complexity of the known method, due to the need to conduct a large array of measurements and their comparison, insufficient reliability of the results obtained by different operators, as well as the inability to maintain the appearance of biological material of different age categories for a long time.

Rapid biochemical changes in the external and internal tissues of the test sample in the early stages of development make it impossible to create a bank of morphological changes associated with the color or structure of certain meristic structural characteristics.

The present invention is aimed at reducing the cost and complexity of assessing the morphological characteristics of aquaculture objects, increasing the reliability of the final result of the morphometric description by eliminating the subjective factor, maintaining the appearance of the biological object for further research for a long time.

The specified technical result is achieved by the fact that in the known method for assessing and preserving the morphometric characteristics of aquaculture objects, including selecting aquaculture objects, fixing them, conducting measurements and saving parameters for further analysis, the feature is that the objects are fixed and their morphometric parameters are saved by instant digital photography of the appearance of objects at the stage under study from the beginning of the division of eggs to age with signs of puberty in fish and anija morphotype computer database characteristics and appearance of aquaculture facilities. Photographing is carried out at optimal illumination at right angles to the object of study against the background of a measuring scale, and morphometric measurements are carried out using the "Ruler" tool of the Photoshop graphics program with an accuracy of at least 0.1 mm, while the linear dimensions of the object are determined by the formula:

X = (a × b) / c,

where: X - the desired value of the linear size of the measured object, mm;

and - the value of the linear size of the measured object, calculated in units of the program "Photoshop";

b - the value of the length of the measuring scale in the photograph, mm;

c is the value of the length of the measuring scale, calculated in units of the Photoshop program.

It is advisable to photograph objects using digital equipment with a resolution of at least 300 dpi in macro mode.

Saving the appearance of objects in the form of a digital photograph is preferably carried out in a computer data bank in JPEG or TIFF format.

Fixing objects by digitally photographing their appearance avoids the conservation of biological objects in fixing media during measurements, reduces the cost and complexity of evaluating the morphological features of aquaculture objects and improves the reliability of the final result.

The creation of a computer database of the characteristics of the morphotype and appearance of aquaculture objects allows us to preserve for a long time for further analysis the appearance of biological objects at different stages of development, without violating the proportions and natural color of the skin.

Carrying out measurements of morphological characteristics with the help of the Ruler tool of the Photoshop graphics program with an accuracy of at least 0.1 mm using the above formula, one can obtain a reliable result of assessing the morphometric characteristics of an object that completely excludes the subjective factor.

It is advisable to photograph objects using digital equipment in macro mode, at a resolution of at least 300 dpi, which allows you to maximize highlight and capture the external and color features of the object.

Saving the image of the appearance of objects in the form of a digital photograph is preferably carried out in a computer database in JPEG or TIFF format, because in these formats digital photo processing in the Photoshop graphics program is more convenient

In the claimed method, errors made by the operator during manipulations with the real object of study are excluded from the analysis. The possibility of repeating the measurement and study of morphological (or phonetic) parameters for many years is respected. Conditions are created for preserving the appearance of biological material in its original form for a long time. The possibility of carrying out comparative ontogenetic and population studies based on a data bank on the collected material is being realized. To obtain a reliable result of the morphological characteristics of objects, a large number of repetitions of measurements is not required, which greatly simplifies and reduces the complexity of describing the morphotype when conducting breeding, breeding and other studies of aquaculture objects.

Thus, the set of distinctive features of the described method ensures the achievement of the specified technical result.

The analysis of the prior art made it possible to establish that the source was not detected, characterized by signs identical to all the essential features of the claimed invention. Therefore, the claimed invention meets the condition of "novelty."

An additional search for known solutions showed that the claimed invention does not follow explicitly from the prior art for a specialist, therefore, the claimed invention meets the condition of "inventive step".

Information confirming the possibility of carrying out the invention with obtaining the above technical result.

The method is as follows.

The objects of study are placed on a solid plane located in a horizontal position and painted in light or dark colors depending on the size and color of the material being studied. Near the objects have a measuring ruler and a label with a description of the subject of study. To photograph the appearance of objects using digital equipment with a resolution of at least 300 dpi. Shooting is performed under optimal illumination in the macro mode of vertical shooting of the surface of the plane in such a way that there are objects of research, a ruler and a label in the photographing area.

The morphometric parameters of objects at different stages of development are fixed and saved by transferring the received information in the form of digital photographs from digital equipment to the magnetic media of a personal computer in a separate folder to create a computer database of digital images of the appearance of aquaculture objects and conduct morphometric measurements in a graphics program . Saving the image in the computer database is preferably carried out in JPEG or TIFF format.

To conduct morphometric measurements, use the ruler tool of the Photoshop graphics program with an accuracy of at least 0.1 mm, while the linear dimensions of the object are determined by the formula

X = (a × b) / c,

where: X - the desired value of the linear size of the measured object, mm;

and - the value of the linear size of the measured object, calculated in units of the program "Photoshop";

b - the value of the length of the measuring scale in the photograph, mm;

c is the value of the length of the measuring scale, calculated in units of the Photoshop program.

The measurement data, the name of the object and the date of fixation of the biological material are transferred to Excel or Statistica spreadsheets and stored in the database.

Thus, a computer database of morphometric characteristics and appearance of the studied object at the required stage of development is created. This allows you to use the aquaculture object for further research with high reproducibility of measurement results. The use of a computer graphics program ensures the complete repeatability of the morphological description of the selected object and its use in comparative studies for many years.

The invention is illustrated by examples.

Example 1. As the object of study was taken caviar (or embryos) of various breeds of carp. Caviar incubation was carried out at the factory in Weiss apparatuses. After straining, a small sample of eggs (approximately 250-300 mg) was taken from the female eggs, it was weighed on a torsion balance, the number of eggs in the taken sample was calculated, and the number of eggs per 1 g was calculated (for further calculation of the female fertility and weight of 1 egg). 24 hours after fertilization, a small arbitrary amount of caviar was taken from the apparatus and placed on a Petri dish with water. A marker was made on the fish breed on the Petri dish, and a measuring scale was placed under the cup to determine the scale when working in the graphics program. A Petri dish with caviar was placed on a solid plane, painted in a dark tone, located in a horizontal position.

Photography was carried out using a digital camera with a resolution of 300 dpi. The shooting was carried out in macro mode without flash flashing vertically on the surface of the plane, while in the photography area were a Petri dish with caviar, a ruler and a label.

The morphometric parameters of caviar were preserved by transferring the photograph on magnetic media of a personal computer to a separate folder to create a database of digital images of the appearance of caviar and to conduct morphometric measurements in a graphics program. Saving the image in the computer database was performed in JPEG format.

To conduct morphometric measurements of eggs, we used the graphic program Photoshop.

After downloading and opening the file with the photo in the window of the graphic editor, its image appears (figure 1). Next, on the tool scale of the program, a button with the "Ruler" icon was selected (in Fig. 1 - A). Measured by "Ruler" the diameter of the required number of carp eggs (figure 1 - C). The measurement result, calculated in units of the program "Photoshop", appears on the top menu of the program (figure 1 - B).

In our case, the diameter of one egg in the units of measurement of the program (a) was 0.41. To convert the units of measure into the metric system, the measuring scale shown in the photograph next to the Petri dish (as shown in the drawing) was used. The measurement was carried out using the Ruler program tool, the distance of a measuring scale equal to 10 mm (b). In the above example, this value of the length of the measuring scale, calculated in units of the program, is 1.26 (s). Based on the data obtained, the egg diameter was calculated by the formula

X = (a × b) / s = (0.41 × 10) / 1.26 = 3.26 mm.

The obtained results of measuring the diameter of the eggs were entered into electronic tables in the EXCEL program to create a data bank and further statistical processing of the morphotype characteristics.

Example 2. As an object of study, larvae of various breeds of carp were taken. Larvae (an arbitrary amount) were caught from the tray on the first day after hatching and after becoming “afloat” (after about 4 days). A cup with water and larvae, due to their weak color, was placed on a white matte background and then operations were carried out analogously to example 1.

After loading and opening the photo in the window of the graphical editor (program "Photoshop"), its image appears (figure 2). Next, on the program tool scale, a button with the “Ruler” icon was selected (in Fig. 2 - A). Measured "Ruler" the body length of the required number of carp larvae (figure 2 - C). The measurement results appear on the top menu of the program (figure 2 - C).

In our case, the length of one larva in the units of measurement of program (a) was 1.51. To convert the units of measure into the metric system, the measuring scale shown in the photograph next to the Petri dish (as shown in the drawing) was used. The measurement was carried out using the Ruler program tool, the distance of a measuring scale equal to 10 mm (b). In the above example, this value of the length of the measuring scale, calculated in units of the program, is 2.23 (s). Based on the data obtained, the length of the larva was calculated by the formula

X = (a × b) / c = (1.51 × 10) / 2.23 = 6.77 mm.

The results obtained by measuring the length of the larvae were entered into electronic tables in the EXCEL program to create a data bank and further statistical processing of the morphotype characteristics.

Example 3. The proposed method was used to study the morphometric characteristics of fish. At the same time, the study of repairs and manufacturers was carried out in vivo, which took about 2-3 minutes. The fish was placed on the wet surface of the sorting table and a label with the necessary data and a ruler were placed next to it. The number of fish on the table can vary depending on age and, accordingly, their size. In our work, the number of fish ranged from 3 to 15 pieces. Next, operations were carried out analogously to example 1.

Morphometric measurements of fish were performed in the graphics program "Photoshop". For example, a digital photograph is shown depicting 10 specimens of two-year-olds of the Moscow scaly carp breed (Fig. 3).

Six morphometric characters were measured for all fish shown in the photograph in the program units (a). To convert the units of measure into the metric system, the measuring scale shown in the photograph (as shown in the drawing) was used. A measurement was made using the Ruler program tool, the distance of a measuring scale equal to 1 cm (b). In the given example, this value of the length of the measuring scale, calculated in units of the program, is 0.30 (c).

Based on the data obtained, the sizes of morphometric characters in cm were calculated using the formula X = (a × b) / c.

The results of measurements are presented in the table.

Table Morphometric description of two-year-old carp of the Moscow Scaly breed Sign Photoshop units (a) The value of the linear size of the measured characteristic (X), cm Body length 9.36 31,20 Body height 3.83 12.76 Head length 2.70 9.00 Head height 1.91 6.37 Caudal stem length 1,50 5.00 Caudal stem height 1.41 4.70

The obtained results of measurements of morphometric characteristics were entered into electronic tables in the EXCEL program to create a data bank and further statistical processing of morphotype characteristics.

Using the proposed method allows for measurement to increase the necessary parts of the body, which makes it possible to more accurately measure the dimensional characteristics of plastic features, as well as calculate the number of scales in the side line or the number of rays in the fins, etc.

Example 4. The proposed method can be used to study not only external but also internal organs, in particular the size and shape of the swimming bladder, intestines and gonads.

All operations are similar to the examples described above (example 1-3).

The example shows the measurement of the swimming bladder in carp (figure 4). The lengths of AB and BC, respectively, determine the length of the posterior and anterior chambers of the swim bladder. The measured values in the units of measure of the program (a) were: AB = 13.09, BC = 19.06, the distance of the measuring scale, equal to 1 cm (b), corresponds to 1.87 units of measurement of the program (s).

Based on the data obtained, the length of the posterior and anterior chambers of the swim bladder was calculated using the formula X = (a × b) / c.

The length of the anterior chamber is (19.06 × 1) / 1.87 = 10.19 cm.

The length of the rear chamber is (13.09 × 1) / 1.87 = 7.00 cm.

The obtained results of measurements of the swimming bladder of carps were entered into spreadsheets in the EXCEL program to create a data bank and further statistical processing.

The above examples illustrate that the claimed method for evaluating and preserving the morphometric characteristics of aquaculture objects allows to obtain a more correct result of the morphometric description of biological material, significantly increase the accuracy and reliability of measurements by eliminating the subjective factor, reduce the complexity and cost of the resulting final product (characteristics of the study object), makes it possible to preserve for a long time the appearance of the biological about an object without disturbing proportions and the natural color of the skin for further research.

Using the method will improve the efficiency of breeding and genetic research aimed at creating and maintaining new breeding achievements, a collection fund of especially valuable fish species, conducting ichthyological and systemological studies of natural populations of ichthyofauna, monitoring changes in aquaculture objects in the development process.

Thus, the above information indicates that when using the claimed invention, the following combination of conditions:

- a method for assessing and preserving the morphometric characteristics of aquaculture objects according to the claimed invention is intended for use in the field of research, monitoring and systematics of biological objects, in particular when conducting breeding and breeding work with aquaculture objects;

- for the claimed method in the form described in the independent clause of the claims, the possibility of its implementation using the means and methods described in the application is confirmed.

Therefore, the claimed invention meets the condition of "industrial applicability".

Claims (3)

1. A method for evaluating and preserving the morphometric characteristics of aquaculture objects, including selecting aquaculture objects, fixing them, conducting measurements and saving parameters for further analysis, characterized in that the objects are fixed and their morphometric parameters are saved by instant digital photographing of objects at the stage being studied from the beginning dividing eggs to age with signs of puberty of fish and creating a computer database of the characteristics of the morphotype and appearance of aquaculture objects, photo shooting is carried out at optimal illumination at right angles to the object of study against the background of a measuring scale, and morphometric measurements are carried out using the "Line" tool of the Photoshop graphics program with an accuracy of at least 0.1 mm, while the linear dimensions of the object are determined by the formula:
X = (a b) / c,
where X is the desired value of the linear size of the measured object, mm;
and - the value of the linear size of the measured object, calculated in units of the program "Photoshop";
b - the value of the length of the measuring scale in the photograph, mm;
c is the value of the length of the measuring scale, calculated in units of the Photoshop program. 2. The method according to claim 1, characterized in that the photographing of objects is carried out using digital equipment with a resolution of at least 300 dpi in macro mode. 3. The method according to claim 1, characterized in that the conservation of the appearance of objects in the form of a digital photograph is preferably carried out in a computer data bank in JPEG or TIFF format.

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