CN106719129B - Method for constructing model for influencing egg yolk utilization rate of pseudobagrus ussuriensis juvenile fish by temperature and photoperiod - Google Patents
Method for constructing model for influencing egg yolk utilization rate of pseudobagrus ussuriensis juvenile fish by temperature and photoperiod Download PDFInfo
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Abstract
The invention relates to a method for constructing a model for influencing the egg yolk utilization rate of a pseudobagrus ussuriensis larva by temperature and a photoperiod, and belongs to the technical field of research on egg yolk utilization of the pseudobagrus ussuriensis larva. The method comprises the following steps: taking the yolk sac larvae of the pseudobagrus ussuriensis which are just hatched out of membranes, and selecting the yolk sac larvae in a normal state as test fish at room temperature; dividing test fish into a plurality of experimental factor combinations, and taking the average volume of yolk sacs of dozens of randomly extracted larvae as initial data before an experiment; when the experiment is finished, randomly selecting 30 larvae from each experiment factor combination, and measuring the egg diameter of the yolk sacs of the larvae in each experiment factor combination; and taking all the obtained yolk utilization rates and the corresponding temperatures and photoperiods thereof, taking the temperatures and the photoperiods as independent variables, taking the yolk utilization rates as dependent variables, and fitting by adopting a least square method to obtain a model of the influence of the temperatures and the photoperiods on the yolk utilization rates of the pseudobagrus ussuriensis juvenile fishes.
Description
Technical Field
The invention belongs to the technical field of research on egg yolk utilization of pseudobagrus ussuriensis larvae, and particularly relates to a method for constructing a model for influencing the egg yolk utilization rate of the pseudobagrus ussuriensis larvae by temperature and photoperiod.
Background
Pseudobagrus ussuriensis (Pseudobagrus ussuriensis) has the advantages of rapid growth, wide eating habits, delicious meat taste and no muscle prickling, and is an excellent top-grade product for cooking and serving as a hot pot material. Pseudobagrus ussuriensis is suitable for pond and net cage culture, has few plant diseases and insect pests, strong market demand and excellent price, and is a promising new species for current aquaculture.
Yolk is a substance gradually accumulated during the growth and development of oocytes, and the main components of the yolk substance of fishes are protein and fat which are necessary nutrient and energy sources for embryonic development and early fry development. Therefore, the consumption and transformation of yolk during the development of embryo and early larval fish directly affect the quality of the new embryo body and the early development of the larval fish, and further affect the growth and survival of the fish. Therefore, the development and nutritional characteristics of the fertilized eggs and the fry are deeply researched, so that the optimal artificial propagation conditions are determined, and the method has important significance on the success of fish seedling production. At present, the research on the yolk metabolism of the fish in the embryonic period is not sufficient at home and abroad, and the research is only seen in the respective yolk utilization aspect of the pelteobagrus vachelli fry and the catfish hybridus at home. Yolk utilization of yellowtail flounder, white sturgeon, herring, striped bass and rainbow trout is reported abroad respectively, but research on the yolk of pseudobagrus ussuriensis is not reported yet.
Disclosure of Invention
The invention aims to solve the defects of the prior art and provides a model for constructing the influence of temperature and light cycle on the egg yolk utilization rate of the pseudobagrus ussuriensis larvae, and the model can provide reliable basis for the breeding quality of the pseudobagrus ussuriensis larvae.
The invention adopts the following technical scheme:
a method for constructing a model for influencing egg yolk utilization rate of pseudobagrus ussuriensis fries by temperature and photoperiod comprises the following steps:
the method comprises the following steps: taking the yolk sac larvae of the pseudobagrus ussuriensis which are just hatched out of membranes, and selecting the yolk sac larvae in a normal state as test fish at room temperature;
step two: taking the test fish obtained in the first step, dividing the test fish into a plurality of experimental factor combinations, measuring the appropriateness of the environment where the larvae are located by using the yolk utilization rate, and taking the randomly extracted average volume of yolk sacs of dozens of larvae as initial data before the experiment; in the experimental process, the temperature and the light period are automatically controlled by an artificial intelligent climate box;
step three: when the experiment is finished, randomly selecting 30 larvae from each experiment factor combination, measuring the egg diameter of the yolk sac of the larvae in each experiment factor combination by using an optical microscope and a micrometer, wherein the egg diameter is a long diameter and a short diameter, and calculating the volume of the yolk sac according to the formula: v-4/3. pi. (r/2)2R/2, where R is the yolk sac minor diameter (mm) and R is the yolk sac major diameter (mm), using the average volume data obtainedPerforming statistical analysis;
the egg yolk utilization at the end of the experiment was calculated according to this equation: pd(％)＝(Vi-Vd)/ViX 100% of formula (II) wherein Pd(%) is the yolk utilization at the end of the experiment, ViYolk sac volume for the start of the experiment, VdYolk sac volume at the end of the experiment;
step four: and (4) taking all the yolk utilization rates obtained in the step three and the corresponding temperature and photoperiod thereof, taking the temperature and photoperiod as independent variables and the yolk utilization rate as dependent variables, and fitting by adopting a least square method to obtain a model of the influence of the temperature and the photoperiod on the yolk utilization rate of the pseudobagrus ussuriensis juvenile fish.
Further, the several combinations of experimental factors in step two were 11 combinations of experimental factors (18 ℃ 24 ℃ 30 ℃) x (0h 12h 24h), wherein the center combination (2412) was repeated 2 times.
Further, each test factor combination in the step two is specifically set as: placing 100 fish for test obtained in the first step into a 2000mL beaker, wherein the beaker contains 1000mL of water, the pH value of the water is 7, and the dissolved oxygen is 4.5mg.L-1Ammonia nitrogen<0.06mg.L-1And setting corresponding temperature and light period in advance, placing the artificial intelligent climate box in which all test factor combinations are the same except that the temperature and the light period are different.
Further, the effect of temperature and photoperiod on yolk utilization of the pseudobagrus ussuriensis is modeled as follows:
Pd(％)＝-386.5491+34.7787T+2.4975P+0.017T×P-0.6463T2-0.0762P2，R20.9865, where T is temperature and P is photoperiod.
Compared with the prior art, the invention has the beneficial effects that:
under the combined action of temperature and photoperiod, the utilization efficiency of the pseudobagrus ussuriensis larva luteus is researched, a model for influencing the yolk utilization rate of the pseudobagrus ussuriensis larva by the temperature and photoperiod is constructed, and a reliable basis is provided for the larva breeding quality of the pseudobagrus ussuriensis: after hatching of fertilized eggs of Pseudobagrus ussuriensis, when the temperature is 26 ℃ and the light cycle is 14h, the egg yolk utilization rate of larvae is highest, after hatching, the larvae are cultivated under the condition, the survival rate of the larvae is highest or the quality of the seedlings is best, the model provides a reliable basis for the cultivation of the larvae of Pseudobagrus ussuriensis under different temperatures and light cycles, and provides a reference for a related Pseudobagrus ussuriensis larva egg yolk utilization rate influence model.
Drawings
FIG. 1 is a graph of the response curve of the combined effect of temperature and photoperiod on yolk utilization;
fig. 2 is a line contour plot of the combined effect of temperature and photoperiod on yolk utilization.
Detailed Description
The present invention will be described in further detail with reference to examples.
It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention.
A method for constructing a model for influencing egg yolk utilization rate of pseudobagrus ussuriensis fries by temperature and photoperiod comprises the following steps:
the method comprises the following steps: taking the yolk sac larvae of the pseudobagrus ussuriensis which are just hatched out of membranes, and selecting the yolk sac larvae in a normal state as test fish at room temperature;
step two: taking the test fish obtained in the first step, dividing the test fish into a plurality of experimental factor combinations, measuring the appropriateness of the environment where the larvae are located by using the yolk utilization rate, and taking the randomly extracted average volume of yolk sacs of dozens of larvae as initial data before the experiment; in the experimental process, the temperature and the light period are automatically controlled by an artificial intelligent climate box;
step three: when the experiment is finished, randomly selecting 30 larvae from each experiment factor combination, measuring the egg diameter of the yolk sac of the larvae in each experiment factor combination by using an optical microscope and a micrometer, wherein the egg diameter is a long diameter and a short diameter, and calculating the volume of the yolk sac according to the formula: v-4/3. pi. (r/2)2R/2, wherein R is the yolk sac minor diameter (mm) and R is the yolk sac major diameter (mm), and statistical analysis is performed by using the obtained average volume data;
the egg yolk utilization at the end of the experiment was calculated according to this equation: pd(％)＝(Vi-Vd)/ViX 100% of formula (II) wherein Pd(%) is the yolk utilization at the end of the experiment, ViYolk sac volume for the start of the experiment, VdYolk sac volume at the end of the experiment;
step four: and (4) taking all the yolk utilization rates obtained in the step three and the corresponding temperature and photoperiod thereof, taking the temperature and photoperiod as independent variables and the yolk utilization rate as dependent variables, and fitting by adopting a least square method to obtain a model of the influence of the temperature and the photoperiod on the yolk utilization rate of the pseudobagrus ussuriensis juvenile fish. Wherein the plurality of combinations of experimental factors in step two is 11 combinations of experimental factors (18 ℃ 24 ℃ 30 ℃) x (0h 12h 24h), wherein the center combination (2412) is repeated 2 times. And each test factor combination in the second step is specifically set as: placing 100 fish for test obtained in the first step into a 2000mL beaker, wherein the beaker contains 1000mL of water, the pH value of the water is 7, and the dissolved oxygen is 4.5mg.L-1Ammonia nitrogen<0.06mg.L-1And setting corresponding temperature and light period in advance, placing the artificial intelligent climate box in which all test factor combinations are the same except that the temperature and the light period are different. And fourthly, the model of the influence of the temperature and the photoperiod on the egg yolk utilization rate of the pseudobagrus ussuriensis fries is as follows:
Pd(％)＝-386.5491+34.7787T+2.4975P+0.017T×P-0.6463T2-0.0762P2，R20.9865, where T is temperature and P is photoperiod.
Example 1
The biological materials used in the test are the same batch of early-hatched pseudobagrus ussuriensis yolk sac larvae, the quantity of the pseudobagrus ussuriensis yolk sac larvae is sufficient, and the biological materials are provided by a test base (a wharf town of Huai-yin area in Huai-yin city, Jiangsu province). Before the experiment, 100 random samples were taken and the long diameter (mm) and short diameter (mm) of the yolk sac were measured using a microscope (power X4), and the average yolk sac volume was calculated according to the formula and used as the initial yolk sac volume at the start of the experiment.
The formula used to calculate the yolk sac volume is: v-4/3. pi. (r/2)2R/2, wherein V (mm)3) The yolk sac volume is shown as R and R are respectively the short diameter and the long diameter (mm) of the yolk sac, and pi is the circumferential rate.
The protocol was designed as in Table 1, with a 2000mL beaker (1000 mL water, pH 7, dissolved oxygen 4.5 mg.L) being placed before the test-1Ammonia nitrogen<0.06mg.L-1) The device is placed in an artificial intelligence climate box (model RTOP-150Y), and the corresponding temperature and the corresponding photoperiod are set in advance. After the water temperature reaches the corresponding setting, 100 egg yolk sacs which are hatched initially are placed in each factor combination, and water body is slightly aerated to keep the larvae in a suspension state. At 96 hours after the start of the experiment, 30 larvae were randomly drawn from each factor combination, the egg diameter was measured microscopically (× 4), and the estimated average yolk sac volume was calculated.
The yolk sac volume was calculated according to this equation: v-4/3. pi. (r/2)2R/2, wherein R is the yolk sac minor diameter (mm) and R is the yolk sac major diameter (mm), and statistical analysis is performed by using the obtained average volume data;
the death phenomenon appears in 4 factor combinations during the test period, and the total death rate is less than or equal to 3 percent.
TABLE 1 combination of factors in the experimental design and corresponding results
The combined effect of two important environmental factors, temperature (T, ° C) and photoperiod (P, h), on the utilization rate of the bursa of Pseudobagrus ussuriensis larvae (Yolk dissolution,%) was examined by using a central composite design. Yolk utilization in yolk sac volume (mm)3) Is shown. According to the result of the preliminary test, the variation ranges of the temperature and the photoperiod are respectively 18 ℃ to 30 ℃ and 0-24 h. By adopting the test design, each test factor respectively takes 3 levels, the number of axis points is 4, the number of factor points is 4, the number of center points is set to be 3, and the length of the star arm is |1 |. The experimental design was designed with 11 factor combinations.
The egg yolk utilization after the start of the experiment was calculated according to this formula: pd(％)＝(Vi-Vd)/Vi×100％
In the formula Pd(%) at the end of the testYolk utilization factor, ViAnd VdYolk sac volumes at the beginning and end of the experiment, respectively.
The experimental error is assumed to follow normal distribution with the mean value of 0, and each effect in the model is estimated by adopting a least square method. The experimental data are processed by SAS (V9.13) statistical software, and the significance and the appropriateness of the model and the significance of the estimation of each effect in the model are tested by an analysis of variance method. The fitting degree of the model is used to determine the coefficient R2And (4) measuring. In order to clearly observe the combined effect of the two test factors on the utilization rate of the yolk sac, the research also provides a response surface graph and a contour graph. The significant level in the test was set to 5%.
As can be seen from the drawings:
1. effect of different temperatures on yolk utilization of Pseudobagrus ussuriensis larvae
As can be seen from fig. 1, the egg yolk utilization rate of the larvae is increased and then decreased along with the change of the temperature, and the egg yolk utilization rate of the larvae is highest at 26 ℃; the primary effect of temperature is very significant (P <0.01) on the yolk utilization of pseudobagrus ussuriensis larvae, and the secondary effect of temperature is very significant (P <0.01) on the yolk utilization of pseudobagrus ussuriensis larvae. The utilization rate of the yolk of the larva is the best at the temperature. Therefore, the water temperature can be kept at about 26 ℃ during the culture process of the pseudobagrus ussuriensis larvae, so that the growth and development speed of the larvae can be improved.
2. Effect of different photoperiods on egg yolk utilization by Pseudobagrus ussuriensis larvae
As can be seen from fig. 1 and 2, both primary and secondary effects of the photoperiod have a significant effect on larval egg yolk utilization (P < 0.05). However, compared with the experimental data under the shortest illumination time, the larval egg yolk utilization rate can be kept at a higher level by long-time uninterrupted illumination, particularly about 14 h. This can provide effective measures for increasing production benefits during seedling cultivation.
3. Interplay of temperature and photoperiod on Pseudobagrus ussuriensis larva luteal utilization
The combined effect test result of the temperature and photoperiod double factors on the utilization rate of the yolk of the larva shows that the interaction between the temperature and the photoperiod is opposite to the utilization rate of the yolk of the pseudobagrus ussuriensisThe effect is not significant (P)>0.05). Obtaining the utilization rate P of the yolk of the larvadThe quadratic mathematical model and the degree of fit for temperature and photoperiod are:
Pd(％)＝-386.5491+34.7787T+2.4975P+0.017T×P-0.6463T2-0.0762P2，R20.9865, where T is temperature and P is photoperiod.
The model can predict the yolk utilization rate under different temperatures and photoperiods, and can obtain the optimal test factor combination through an optimization technology.
Through the studies of the present application, the following conclusions were drawn:
1) the egg yolk utilization rate of pseudobagrus ussuriensis larvae is closely related to environmental factors, and the appropriate temperature and photoperiod have a remarkable promoting effect on the egg yolk utilization of the larvae. The utilization rate of the yolk of the larva presents a curve change trend along with the temperature and the photoperiod;
2) there is no interaction between the two environmental factors temperature and photoperiod;
3) a quadratic mathematical model of the egg yolk utilization rate of the pseudobagrus ussuriensis larvae on temperature and photoperiod is established, and the fitting degree of the model is as high as 98.65%;
4) the result shows that the larva egg yolk utilization rate reaches the highest at the factor combination position of 26 ℃/14h, and the method has great practical guiding significance.
It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (4)
1. A method for constructing a model for influencing egg yolk utilization rate of Pseudobagrus ussuriensis larvae through temperature and photoperiod, which is characterized by comprising the following steps:
the method comprises the following steps: taking the yolk sac larvae of the pseudobagrus ussuriensis which are just hatched out of membranes, and selecting the yolk sac larvae in a normal state as test fish at room temperature;
step two: taking the test fish obtained in the first step, dividing the test fish into a plurality of experimental factor combinations, measuring the appropriateness of the environment where the larvae are located by using the yolk utilization rate, and taking the randomly extracted average volume of yolk sacs of dozens of larvae as initial data before the experiment; in the experimental process, the temperature and the light period are automatically controlled by an artificial intelligent climate box;
step three: when the experiment is finished, randomly selecting 30 larvae from each experiment factor combination, measuring the egg diameter of the yolk sac of the larvae in each experiment factor combination by using an optical microscope and a micrometer, wherein the egg diameter is a long diameter and a short diameter, and calculating the volume of the yolk sac according to the formula: v-4/3. pi. (r/2)2R/2, wherein R is the yolk sac minor diameter, mm; r is the length diameter of the yolk sac, mm; performing statistical analysis by using the obtained average volume data;
the egg yolk utilization at the end of the experiment was calculated according to this equation: pd(％)＝(Vi-Vd)/ViX 100% of formula (II) wherein Pd(%) is the yolk utilization at the end of the experiment, ViYolk sac volume for the start of the experiment, VdYolk sac volume at the end of the experiment;
step four: and (4) taking all the yolk utilization rates obtained in the step three and the corresponding temperature and photoperiod thereof, taking the temperature and photoperiod as independent variables and the yolk utilization rate as dependent variables, and fitting by adopting a least square method to obtain a model of the influence of the temperature and the photoperiod on the yolk utilization rate of the pseudobagrus ussuriensis juvenile fish.
2. The method for modeling the influence of temperature and photoperiod on the yolk utilization of the pseudobagrus ussuriensis according to claim 1, wherein said plurality of experimental factor combinations in step two are 11 experimental factor combinations: (18 ℃, 24 ℃, 30 ℃) x (0h, 12h, 24h), center point combination: repeat 2 times at 24 ℃ for 12 h.
3. The method of claim 1, wherein the influence of temperature and photoperiod on egg yolk utilization of Pseudobagrus ussuriensis is modeled in step twoEach experimental factor combination is specifically set as: placing 100 fish for test obtained in the first step into a 2000mL beaker, wherein the beaker contains 1000mL of water, the pH value of the water is 7, and the dissolved oxygen is 4.5mg.L-1Ammonia nitrogen<0.06mg.L-1And setting corresponding temperature and light period in advance, placing the artificial intelligent climate box in which all test factor combinations are the same except that the temperature and the light period are different.
4. The method for modeling the influence of temperature and photoperiod on the yolk utilization rate of the pseudobagrus ussuriensis, as claimed in claim 1, wherein the model of the influence of temperature and photoperiod on the yolk utilization rate of the pseudobagrus ussuriensis is as follows:
Pd(％)＝-386.5491+34.7787T+2.4975P+0.017T×P-0.6463T2-0.0762P2，R20.9865, where T is temperature and P is photoperiod.
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