CN111657196A - Macrobrachium rosenbergii larva breeding method - Google Patents
Macrobrachium rosenbergii larva breeding method Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/50—Culture of aquatic animals of shellfish
- A01K61/59—Culture of aquatic animals of shellfish of crustaceans, e.g. lobsters or shrimps
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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Abstract
The invention belongs to the technical field of aquaculture, and discloses a macrobrachium rosenbergii larva culturing method, which analyzes the influence of three environmental factors, namely 3 salinity, 5 photoperiods, 5 visible light irradiation and the like, on the survival, metamorphosis, growth and related enzyme activities of macrobrachium rosenbergii larvae. The survival rate, metamorphosis rate, growth and related enzyme activities of the macrobrachium rosenbergii larvae cultured at different salinity are all obviously different, wherein the experimental group with the salinity of 13 per mill has the best growth, development and survival conditions; under different illumination periods, the survival, metamorphosis, growth and related enzyme activities of the larvae of the macrobrachium rosenbergii are obviously different, and the survival, metamorphosis and growth effects of the larvae are optimal under the continuous illumination condition; under different visible light irradiation, the metamorphosis, survival and growth effects of the macrobrachium rosenbergii larvae under the irradiation of white light and green light are the best, and the growth, development and survival indexes of the red light experimental group are the worst. In the culture production, the seedling raising effect of the macrobrachium rosenbergii can be improved by proper salinity, optimal photoperiod and visible light.
Description
Technical Field
The invention belongs to the technical field of aquaculture, particularly relates to a macrobrachium rosenbergii larva breeding method, and particularly relates to a breeding method for influencing survival, growth and related enzyme activity indexes of macrobrachium rosenbergii larvae by salinity, photoperiod and visible light.
Background
At present, Macrobrachium rosenbergii (Macrobrachium rosenbergii) is originally produced in southeast Asia, and is gradually one of the main shrimp varieties for artificial freshwater aquaculture in China since the last 70 th century when being introduced into China. According to the statistics of fishery yearbook, the total yield of the macrobrachium rosenbergii in 2018 reaches 13.3 million tons. Compared with other shrimp varieties, the main reason for restricting the increase of the culture yield of the macrobrachium rosenbergii is the insufficient supply of high-quality seedlings besides the reasons of consumption markets, wherein the low emergence rate is one of important reasons.
Therefore, how to improve the metamorphosis of the larvae and the survival of the shrimp larvae is the central importance of the current macrobrachium rosenbergii industry for breaking through the development bottleneck. The water quality plays a decisive role in the growth and survival conditions of the cultured macrobrachium rosenbergii, and although the macrobrachium rosenbergii is a freshwater cultured variety, the larva hatched from fertilized eggs needs a certain salinity to normally grow and develop; meanwhile, the photoperiod and the optical wavelength also have certain influence on the growth and development of the shrimps. But this is a sporadic study.
Through the above analysis, the problems and defects of the prior art are as follows:
(1) the prior art does not systematically analyze the influence of different salinity, photoperiod and visible spectrum on the metamorphosis, survival, growth and related enzyme activity indexes of the macrobrachium rosenbergii larvae, and breeding enterprises fight against each other and lack corresponding breeding standards, so that the growth and development rate of the macrobrachium rosenbergii larvae is low, and the technical effect of the macrobrachium rosenbergii seedling breeding is poor.
(2) In the process of supplying and changing water, the salinity change is not strictly controlled by operators, so that stress to a certain degree is caused, and the survival of the larva is influenced.
(3) The cultivation personnel are not aware of the importance of the illumination, can not provide sufficient long-term illumination for the larva, and can not provide the optimal illumination condition for the growth and development of the larva.
The difficulty in solving the above problems and defects is:
(1) the optimum salinity, photoperiod and visible spectrum of the growth and development of the larvae of the macrobrachium rosenbergii are not confirmed, and special culture experiments are needed for confirmation.
(2) The requirement is strict during actual operation, the staff is required to be trained and have accountability, and the equipment is arranged in place.
The significance of solving the problems and the defects is as follows: the method scientifically analyzes the influence of different salinity, photoperiod and visible light on the metamorphosis, survival, growth and related enzyme activity indexes of the macrobrachium rosenbergii larvae, finds out ecological factors such as salinity, photoperiod and visible light which are most suitable for the growth and development of the macrobrachium rosenbergii larvae, and provides guidance for the optimization of the macrobrachium rosenbergii seedling culture technology. Can provide the optimal production management standard for the production of the Luo's shrimp seeds and promote the industrial specialized systematization.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a method for culturing the larvae of the macrobrachium rosenbergii.
The invention is realized in such a way that a macrobrachium rosenbergii larva breeding method comprises the following steps:
and (3) salinity analysis: designing 16 per mill of high salinity group, 13 per mill of medium salinity group and 10 per mill of low salinity group; each group is provided with three parallels;
and (3) illumination analysis: all experimental barrels and internal light sources are covered by light-tight black mulching films, and LED bulbs with various colors are arranged right above the barrels; all the power supply switching time is controlled by an automatic timer, and each group is provided with three parallels;
and (3) photoperiod analysis: designing 5 groups of experiment groups with different photoperiods, and irradiating by using white light, wherein the irradiation time is 0:24-h,6:18-h,12:12-h,18:6-h and 24:0-h, and each group is parallel to three;
visible light analysis: different groups of 5 visible light experimental groups are designed, the wavelengths of the visible light are red (615 nm-650 nm), yellow (580 nm-595 nm), white (450 nm-465 nm), green (495 nm-530 nm) and blue (450 nm-480 nm), each group is parallel, and the illumination period is 12: 12-h.
Wherein, all experimental culture time are 23 days, the fairy shrimp is fed in the whole daily process, the water quality change (nitrite, dissolved oxygen, temperature, residual chlorine and the like) is monitored every day, and meanwhile, the fifth day is started to absorb the sewage by a siphon method every day to keep the water quality stable;
in the illumination analysis, all experimental barrels and internal light sources are covered by opaque black agricultural mulching films, 35W of LED bulbs with various colors are selected as the light sources and are placed at a position 30cm above the barrels, and the salinity of the culture water is 13 per thousand;
and finally, performing data analysis after finishing the culture result and the enzyme activity result obtained by the experiment, processing data by using single-factor variance analysis, and performing multiple comparison on the mean value by using an LSD (least squares) method, wherein P <0.05 indicates that significant difference exists, and the data is represented by mean value +/-standard deviation (mean +/-SD).
By combining all the technical schemes, the invention has the advantages and positive effects that:
in the experiment of the invention, the macrobrachium rosenbergii larvae under the conditions of salinity of 13 per mill, 24-hour illumination and white light or green light illumination show the best growth, development and survival conditions in the respective experimental groups. Similarly, in the culture production, the optimum salinity, illumination time and visible spectrum found by the invention are used for culturing the macrobrachium rosenbergii, so that the culture effect of the macrobrachium rosenbergii under the same condition can be obviously improved. Meanwhile, the method is economical and practical, convenient to operate and regulate and can be directly and easily applied to actual production.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings needed to be used in the embodiments of the present application will be briefly described below, and it is obvious that the drawings described below are only some embodiments of the present application, and it is obvious for those skilled in the art that other drawings can be obtained from the drawings without creative efforts.
FIG. 1 is an indicator diagram of enzyme activity of macrobrachium rosenbergii larvae at different salinity according to the embodiment of the invention.
FIG. 2 is an indicator diagram of the enzyme activity of the larvae of Macrobrachium rosenbergii at different photoperiods according to the present invention.
Fig. 3 is an indicator diagram of the enzyme activity of the macrobrachium rosenbergii larvae under different wavelengths of visible light provided by the embodiment of the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Aiming at the problems in the prior art, the invention provides a method for culturing the larvae of the macrobrachium rosenbergii, and the invention is described in detail with reference to the attached drawings.
The macrobrachium rosenbergii larva breeding method provided by the invention comprises the following steps:
and (3) salinity analysis: designing 16 per mill of high salinity group, 13 per mill of medium salinity group and 10 per mill of low salinity group; each set being arranged in a plurality of parallel.
And (3) illumination analysis: all experimental barrels and internal light sources are covered by light-tight black mulching films, and LED bulbs with various colors are arranged right above the barrels; all power supply switching time is controlled by an automatic timer, and each group is provided with a plurality of parallels.
And (3) photoperiod analysis: 5 groups of experiment groups with different photoperiods are designed, white light is used for irradiation, and the irradiation time is 0:24-h,6:18-h,12:12-h,18:6-h and 24:0-h respectively.
Visible light analysis: designing different groups 5 of visible light experimental groups, wherein the visible light color wavelengths are red (615 nm-650 nm), yellow (580 nm-595 nm), white (450 nm-465 nm), green (495 nm-530 nm) and blue (450 nm-480 nm); the illumination period is 12: 12-h.
In the invention, 13 per mill of medium salinity groups are adopted in salinity analysis, and 3 groups are arranged in parallel.
In the illumination analysis, all experimental barrels and internal light sources are covered by light-tight black agricultural mulching films, and 35W LED bulbs with various colors are selected as the light sources and are arranged at the position 30cm above the barrels;
each group is provided with 3 parallels; the salinity of water for each cultivation is 13 per mill.
The salinity analysis, the illumination analysis, the photoperiod analysis and the visible light analysis are carried out as follows:
and (3) data analysis, namely, the experimental data are sorted and analyzed, the data are processed by single-factor variance analysis, the LSD method mean value is subjected to multiple comparison, the P <0.05 shows that the significant difference exists, and the data are shown as the mean value +/-standard deviation (mean +/-SD).
The macrobrachium rosenbergii larva breeding method provided by the invention can be implemented by other steps by persons skilled in the art, and the method is only one specific embodiment.
The invention is further described below in connection with specific experiments.
1. Materials and methods
1.1 Experimental materials
All experiments were carried out in 2019 in san shui Cui pit platinum Miao GmbH, Fushan City.
The experimental materials for the experimental shrimp are all from the same batch of hatched macrobrachium rosenbergii larva groups in the same hatching pond.
The experimental water is culture seawater which is transported from the pearl sea to the Buddha mountain and is disinfected, deposited in a dark room and fully aerated, and the salinity is 13-14 per mill. The salt used for increasing the salinity is the sun salt produced in Shandong sunshine.
1.2 Experimental methods
1.2.1 Experimental design
The single seedling raising time of the Macrobrachium nipponensis culture enterprises in Guangdong province is generally 20-25 days, the seedlings can emerge when the metamorphosis juvenile shrimps account for 40% -50% of the total amount, and the culture time of all experiments is set to be 23 days according to actual production experience.
The salinity experiment design is 16 per mill of high salinity group, 13 per mill of medium salinity group and 10 per mill of low salinity group. Each set was set to 3 replicates. The macrobrachium rosenbergii larvae in each group are 2 thousands of tails.
All experimental barrels and internal light sources of the illumination experiment are covered by light-tight black agricultural mulching films, no influence of external light sources is guaranteed, and 35W LED bulbs with various colors are selected as the light sources and are arranged at the positions 30cm above the barrels. All power on-off time is controlled by a product benefit AL-06 automatic timer, and 3 parallels are arranged in each group. The macrobrachium rosenbergii larvae in each group are 2 ten thousand tails. The salinity of the culture water is 13 per mill.
The photoperiod designs of 5 groups of different photoperiod experimental groups are irradiated by white light, and the illumination time (L) and the non-illumination time (D) are respectively 0:24-h,6:18-h,12:12-h,18:6-h and 24: 0-h.
The visible light designs are different from the 5 groups of the visible light experimental groups, and the colors (wavelengths) of the visible light are respectively red (615 nm-650 nm), yellow (580 nm-595 nm), white (450 nm-465 nm), green (495 nm-530 nm) and blue (450 nm-480 nm). The illumination period is 12: 12-h.
1.2.2 Experimental procedures
Before the experiment, 10 round white barrels with the capacity of 700 liters are prepared, cleaned and disinfected, and then the same culture seawater is added, and the salinity experiment needs to adjust the seawater salinity by fully aerated fresh water and solar salt. Controlling the water level at 50cm, adding clear water modifier into the Zhujiang pond according to the proportion of 2 tablets per 3 experimental barrels, scrubbing the disinfected air with chlorine, aerating, measuring residual chlorine one by one after 24 hours, and adding the larvae for experiment until the residual chlorine does not exceed 0.005. The same seawater for cultivation is added into the redundant 1 white barrel, and aeration is carried out by using the air exhaust for later-stage water adding and water distribution.
And (3) putting the experimental larvae into each group, randomly selecting 2 million larvae of the macrobrachium rosenbergii, putting the 2 million larvae into a barrel, putting an inflatable packaging bag filled with the larvae into the barrel in advance before putting, enabling the water temperature and the water in the barrel to be the same, and preventing stress caused by overlarge temperature difference.
The method comprises the following steps of continuously inflating and oxygenating in a daily management experiment period, feeding fairy shrimp (Artemia) the next day after larvae are thrown in, feeding a small amount of fairy shrimp (Artemia) for many times every day, and controlling the ratio of the larvae to the fairy shrimp in a water body to be 1: 5 or so, and feeding the feed for the last time in a proper amount every day. And (5) beginning on the fifth day, performing dirt suction once every 10 am, adding fresh water with the same salinity as the original barrel to the original water level after dirt suction, and appropriately supplementing the fairy shrimp. To promote normal metamorphosis of the larvae, water was added every 5 days to raise the water level by 5 cm. 30 minutes at 5 pm every day, and each experimental barrel is respectively sampled by a special plastic bottle for water quality detection. The used instrument of water quality testing is octadex brand W-II type water quality testing ware and supporting reagent thereof, and the measurement index has: pH value, ammonia nitrogen, nitrite, dissolved oxygen, residual chlorine, sulfide and total alkalinity, and simultaneously, an illumination meter of the type TA8121 Teansi brand is used for measuring the external temperature and the illumination intensity during sampling.
And (3) collecting and measuring samples, namely estimating the survival amount of the shrimp fries in the experimental barrel by using a five-point sampling method under the conditions that the samples are placed in the mountains in the sun every day, no light source is available, and the distribution of the larvae is uniform, and continuing until the experiment is finished. On day 23, random samples were taken from different locations of the cultivation bucket using 400mL beakers, and 3 beakers were taken each time to calculate an average value. After counting survival rate and metamorphosis rate, 50 tails of the animals were randomly selected from each group, and body length and body weight were measured one by one. Body length was measured using the software tpsdig2 and body weight was measured using a precision balance model AL-204 mettler-toledo. And finally, sampling from the rest shrimp larvae, and performing biochemical index measurement. The biochemical indexes are selected from superoxide dismutase (SOD), Catalase (CAT), Lysozyme (LZM) and alkaline phosphatase (AKP), and are measured by using a kit produced by Nanjing Biochemical Limited.
1.3 data analysis
The experimental data were collated and analyzed using Microsoft Excel 2019 and IBM SPSS Statistics 21, the data were processed by one-way ANOVA, multiple comparisons of the mean values by the LSD method, P <0.05 indicating significant differences, and the data are expressed as mean + -standard deviation (mean + -SD).
2. Results
2.1 Effect of different salinity on survival, growth and related enzyme activity indexes of Macrobrachium rosenbergii larvae
2.1.1 Effect of different salinity on survival and growth of Macrobrachium rosenbergii larvae
The indexes of metamorphosis, survival and growth of each group of macrobrachium rosenbergii larvae after being fed for 23 days at different salinity are shown in table 1. The results show that the survival rate and the metamorphosis rate of the salinity test groups with 10 per mill and 13 per mill are obviously higher than those of the salinity test group with 16 per mill (P < 0.05); the metamorphosis rate and body length of the salinity 13 per mill experimental group are obviously higher than those of the salinity 10 per mill experimental group (P < 0.05). The salinity is proved to have an important effect on the survival and metamorphosis of the larvae of the macrobrachium rosenbergii, and the survival and metamorphosis of the larvae of the macrobrachium rosenbergii can be influenced by overhigh or overlow salinity. The survival, metamorphosis and growth of the macrobrachium rosenbergii larvae cultured under the salinity of 13 per mill are the best.
TABLE 1 growth index of Macrobrachium rosenbergii larvae at different salinity
The letters in the same row are different and represent significant differences (p < 0.05); the letters are identical, meaning that the differences are not significant (p > 0.05); a is the maximum value (the same applies to the table).
2.1.2 Effect of different salinity on the enzyme activity index of macrobrachium rosenbergii larvae
The enzyme activity results of the macrobrachium rosenbergii larvae after being fed for 23 days at different salinity are shown in figure 1. Significant differences occurred in all four measurements for the three salinity groups (P < 0.05): the SOD activity gradually increases from low salinity to high salinity, and the difference between groups is obvious (P is less than 0.05); CAT and AKP activity of a salinity 13 per mill experimental group is obviously higher than 10 per mill and 16 per mill experimental group (P <0.05), and CAT activity of a salinity 10 per mill experimental group is obviously higher than that of a salinity 16 per mill experimental group (P < 0.05); no significant difference in AKP activity between 10 and 16% o experimental groups (P > 0.05); the LZM viability was not significantly different for the 13 and 16% salinity groups (P >0.05), while significantly higher than the 10% salinity group (P < 0.05).
2.2 Effect of different photoperiods on survival, growth and related enzyme activity indexes of Macrobrachium rosenbergii larvae
2.2.1 Effect of different photoperiods on survival and growth of Macrobrachium rosenbergii larvae
The results of the metamorphosis, survival and growth indexes of the macrobrachium rosenbergii larvae cultured in different photoperiods are shown in table 2. The survival and metamorphosis effects of the 0h experimental group are the worst, and the difference with the rest groups is obvious (P is less than 0.05); the survival rate of the larvae of the macrobrachium rosenbergii is not obviously different between the experimental groups of 12h and above (P is more than 0.05), but is obviously higher than that of the experimental groups of 0h and 6h (P is less than 0.05); the allergy rate, body length and body weight of the 24h experimental group were significantly higher than those of the other experimental groups (P < 0.05). The result shows that the illumination is necessary for the growth and development of the larvae, the growth and development conditions of the larvae of the macrobrachium rosenbergii are obviously different under different illumination periods, and the survival, metamorphosis and growth effects of the larvae are optimal under the conditions of continuous illumination and experimental illumination intensity.
TABLE 2 Macrobrachium rosenbergii growth index under different photoperiods
2.2.2 Effect of different photoperiods on the enzyme activity indexes related to the larvae of Macrobrachium rosenbergii
The relative enzyme activities of each group after feeding macrobrachium rosenbergii larvae for 23 days under different photoperiods are shown in figure 2. The SOD activity of each group is reduced along with the increase of illumination time, the difference between the 0h illumination group and the 6h illumination group is not significant (P is more than 0.05), and the difference between other experimental groups is significant (P is less than 0.05); the trend of CAT activity is the same as that of SOD on the whole, but the 0h illumination group is obviously higher than that of other experimental groups (P <0.05), and the CAT activity among other experimental groups is not obviously different (P > 0.05); the AKP activity was significantly higher in the 0h and 6h light groups than in the remaining 3 experimental groups (P <0.05), with no significant difference between them (P >0.05), the lowest and significant AKP activity in the 18h light group (P <0.05), and no significant difference between the 12h and 24h light groups (P > 0.05); the LZM activity change and the AKP general change trend are the same, the LZM activity of the 0h illumination group is remarkably higher than that of the 12h, 18h and 24h experiment groups (P <0.05), the 6h experiment group and all the rest experiment groups have no remarkable difference (P >0.05), and the 12h, 18h and 24h experiment groups have no remarkable difference (P > 0.05).
2.3 Effect of different visible lights on survival, growth and related enzyme activity indexes of Macrobrachium rosenbergii larvae
2.3.1 Effect of different visible lights on survival and growth of Macrobrachium rosenbergii larvae
The metamorphosis, survival and growth indexes of the macrobrachium rosenbergii larvae cultured under different visible lights are shown in table 3. All indexes of the red light group are worst and have obvious difference with other light color experimental groups (P < 0.05). There was no significant difference between survival and metamorphosis between the green, blue, white and yellow light groups (P > 0.05). There was no significant difference between the body lengths of the green and white light groups (P >0.05), but both were significantly higher than the blue and yellow light groups (P < 0.05). In terms of body weight, green light is significantly higher than yellow and red light (P <0.05), but has no significant difference from blue and white light (P > 0.05). The results show that except that red light can obviously inhibit the growth of the larvae of the macrobrachium rosenbergii, the survival rate and the metamorphosis rate of other groups do not reach obvious levels, but the body length and the body weight of the macrobrachium rosenbergii have certain difference among different light colors, and the metamorphosis, survival and growth effects of the larvae of the macrobrachium rosenbergii under the irradiation of white light and green light are optimal. Red (615 nm-650 nm), yellow (580 nm-595 nm), white (450 nm-465 nm), green (495 nm-530 nm), blue (450 nm-480 nm)
TABLE 3 growth index of Macrobrachium rosenbergii under different wavelengths of visible light
2.3.2 Effect of different visible light on enzyme activity indexes related to Macrobrachium rosenbergii larvae
The results of the enzyme activities of the larvae of the macrobrachium rosenbergii are shown in figure 3 when the larvae are raised for 23 days under different visible lights. In SOD activity, yellow color group is significantly higher than other color group (P <0.05) from high to low histogram; the red group was not significantly different from the blue group (P >0.05), but significantly higher than the green and white groups (P < 0.05); the blue light group was significantly higher than the green light group (P <0.05), and there was no significant difference from the white light group (P > 0.05). CAT activity histogram trends were similar to SOD activity, with no significant difference between the yellow and red (P >0.05) and significantly higher than the green and white (P < 0.05); there was no significant difference between the green and white light groups (P >0.05) and no significant difference between the blue and the remaining 4 groups (P > 0.05). The AKP activity was also significantly higher in the yellow and red groups than in the remaining groups (P <0.05) and there were no differences between groups; the white light group is significantly higher than the blue and green light groups (P < 0.05). In LZM activity, the yellow group was significantly higher than the blue, green, and white groups (P < 0.05); the red group is significantly higher than the blue and green groups (P <0.05), but has no significant difference from the yellow and white groups (P > 0.05); the white light group and the blue and green light groups have no significant difference (P > 0.05).
3. The present invention is further described below with reference to the results.
3.1 influence of salinity on growth, survival and related enzyme activity indexes of Macrobrachium rosenbergii larvae
The experimental result shows that the optimum metamorphosis and survival salinity of the macrobrachium rosenbergii larvae are 10-13 per mill. The results of the salt content short-term stress experiments of the macrobrachium rosenbergii larvae carried out by Chenweng et al show that the survival rate of the macrobrachium rosenbergii larvae is the highest in the salt content of 10-14 per mill (gradient 2 per mill), and the results are consistent with the experimental results. The research of You-Hui Huang shows that salinity influences the growth and development of animals with wide salinity by changing osmotic pressure, the animals have the capacity of regulating and adapting to the osmotic pressure, the regulating capacity is limited, and the development of the animals can be enhanced to a certain extent along with individual metamorphosis. In the experimental daily culture process, the macrobrachium rosenbergii in high salinity has the phenomena of slow feeding and reduced activity compared with other treatment groups. The Soeiro et al study compared the survival time of two groups of amazon macrobrachium amansii larvae from different water areas at different salinity without feeding at all, and the results show that: the survival time of the amazon macrobrachium amazonii larvae living in the estuary water area is longest and the vitality is strongest when the salinity is 10; when the salinity is 5, the metamorphosis rate of estuary larvae is 0, and larvae growing in a flooding plain can normally metamorphosis and grow; chand et al performed survival and growth analysis of macrobrachium rosenbergii larvae in different salinity.
The analysis finds that: in a culture environment with the salinity of 0-15 per mill, the survival and growth indexes of the juvenile macrobrachium rosenbergii are not obviously different, but the survival rate of the juvenile macrobrachium rosenbergii is obviously reduced in an environment with the salinity of 20 per mill. Through various analyses, it can be found that the optimum salinity of the same species inhabiting different geographical positions of the same species at different development stages of different species and the same species varies due to different original living water areas and physiological structures. The optimum salinity of the macrobrachium rosenbergii larvae in different metamorphosis development stages has certain difference, and the optimum salinity of each molting period of the larvae is to be found out. According to the existing results, in the current macrobrachium rosenbergii seedling production, the salinity of the water for seedling culture is controlled within the range of 10-13 per mill as much as possible.
In crustaceans, SOD and CAT activities are important indicators for measuring the antioxidant capacity of the body. SOD and CAT have synergistic effect, and can eliminate free radicals generated in organism due to stress by converting hydrogen peroxide into water, and has strong resistance and defense function on oxidative damage of organism cells. In this experiment, the trends of SOD and CAT along with the salinity change are different: the SOD gradually rises, the CAT is firstly increased and then reduced, the difference among the groups is obvious, the CAT activity of the salinity 16 per mill group is too low, the CAT cannot effectively cooperate with the SOD, and the normal mechanism process cannot be carried out, which indicates that the oxidation resistance of the macrobrachium rosenbergii larvae is limited when the salinity is high. AKP is an important index for determining whether the organism is normal and healthy in metabolism and controlling the phosphate group in vivoMetastasis and metabolism[13]In the experiment, the AKP activity of the experiment group with the salinity of 13 per mill is obviously higher than that of other treatment groups, so that the metabolism and the development of the macrobrachium rosenbergii cultured under the salinity of 13 can be presumed to be more positive. The LZM is an alkaline protein, and the stability of the organism is protected mainly by hydrolyzing cell walls of gram-positive bacteria and removing external harmful substances invading the organism, in the experiment, the activity of the LZM in two groups with higher salinity of 13 per mill and 16 per mill is obviously higher than that of a group with low salinity, but the salinity experiment of Zhaoyuqiao on post shrimp of litopenaeus vannamei shows that the LZM has no obvious difference under each salinity gradient. Whether it is a species difference or not requires further experimental confirmation. The SOD enzyme activity range is 95.09U mgprot when the salinity for culturing the larva is within the range of 10-13 ‰-1-105.57U·mgprot-1The range of CAT enzyme activity is 0.85U mgprot-1-1.18U·mgprot-1The AKP enzyme activity range is 70.52U mgprot-1-87.86U·mgprot-1The LZM enzyme activity ranged from 19.02U mgprot-1-41.07U·mgprot-1。
3.2 Effect of photoperiod on growth, survival and related enzyme activity indexes of Macrobrachium rosenbergii larvae
The result of the invention shows that the growth and development index of the macrobrachium rosenbergii larvae is in positive correlation with the illumination time, and the metamorphosis, survival and growth effects of the macrobrachium rosenbergii larvae in a 24-hour illumination group are optimal. 4 gradient photoperiod experiments are designed on macrobrachium rosenbergii larvae before the Takao et al, and researches show that the survival rate of a continuous illumination experiment group is improved by 15.7 percent compared with that of a 4h illumination experiment group, the average total days for breeding the macrobrachium rosenbergii is 23 days, and the average total days is 10 days earlier than that of the 4h illumination experiment group, and the results are consistent with the experiment. In natural environment, the seasonal variation and the photoperiod variation have close relationship, and the living rhythms of the crustaceans distributed in different regions are obviously different, so that the crustaceans have various requirements on illumination. For example, the growth and development of the penaeus japonicus or the Penaeus pinipes are not influenced in different lighting periods; under laboratory conditions, there was no significant difference in the rate of weight gain and rate of growth of the whole black and full exposure groups of procambarus clarkii; meanwhile, Forster finds that the continuous dark environment can promote the growth of the shrimps with the arms; the juvenile macrobrachium rosenbergii grows well in complete darkness, the macrobrachium rosenbergii in the larval stage in the experiment grows better as the illumination time is longer, the different development stages have obvious differences, and the illumination is estimated to promote the metamorphosis of the larval. The effect of photoperiod on crustaceans varies not only from species to species but also with respect to different growth stages of the same species. For the macrobrachium rosenbergii offspring seed enterprises, the illumination time can be increased as much as possible during the larva cultivation period to promote metamorphosis so as to improve the production effect.
Under different photoperiods, the SOD and CAT histograms have consistent trend of gradually reducing activity along with the increase of illumination time, Nelson and other researches find that the oxygen consumption rate of the Macrobrachium rosenbergii under dark condition is 12% higher than that under illumination condition, and the reasons of higher activity of SOD and CAT in the non-illumination group and the 6 h-illumination group can be the frequent oxygen metabolism of shrimp bodies and the high activity of O2 -The excitation effect of a large amount of generated poisons on the organism shows that the larvae need certain illumination to ensure the normal antioxidant system of the organism, and meanwhile, the SOD activity of a continuous illumination group is the lowest, which shows that the illumination for a long time can influence the antioxidant performance of the larvae of the macrobrachium rosenbergii, and the result is similar to the illumination experiment result of the giant ludwigia Dunningtchii on the procambarus clarkii. AKP and LZM activities show higher activities in both the 0h experiment group and the 6h experiment group, and have obvious difference with other experiment groups. The growth and development of the larvae of the macrobrachium rosenbergii are influenced when no light or low-intensity light is indicated, and the enzymatic activities of SOD, CAT, AKP and LZM of the larvae are in a high level under the state; when the larvae are illuminated for a long time, the growth and development of the larvae are better, and the enzyme activities of SOD, CAT, AKP and LZM are in low positions.
3.3 Effect of visible light on growth, survival and related enzyme activity indexes of Macrobrachium rosenbergii larvae
The metamorphosis, survival and growth effects of the larvae of the macrobrachium rosenbergii are the worst under the irradiation of red light, and other lights have no obvious difference. In production, white light which accords with human vision habits is recommended to be selected to supplement the light for the larvae of the macrobrachium rosenbergii. The light wavelength can influence the growth and development of the crustacean by adjusting the hormone secretion, the movement frequency and the breathing consumption of the crustacean, but different species and the same species have certain differences in different development periods, which are closely related to the photosensitive system of the crustacean. The Nagano and the like carry out backwashing on various crustacean by different spectrum irradiation, and the result shows that some types are insensitive to red light and sensitive to blue light; some species are sensitive to different spectrums in different development stages and have different photosensitive systems, for example, two experiments carried out on procambarus clarkii by Fanjul-Moles show that the photosensitive system of the procambarus clarkii in the early development stage is sensitive to blue light, and the photosensitive system of the procambarus clarkii in the later development stage is sensitive to red light. Whether similar multi-photosensory systems exist in macrobrachium rosenbergii is not clear at present, and the macrobrachium rosenbergii is still to be verified by subsequent experiments.
There were more differences between the various enzyme activities of the different visible light treatment groups. In the red light group which is not suitable for metamorphosis, survival and growth of the macrobrachium rosenbergii larvae, the activity of the 4 enzymes is higher and basically consistent with the enzyme activity without illumination, and the growth, metamorphosis and survival results of the larvae without illumination are similar to the results of the red light group, which shows that the red light effect and the dark effect are the same. Probably because of O accumulated in the larvae of the macrobrachium rosenbergii under the irradiation of red light2 -Larger than the other processing groups. The light color experiment of the modern optical cement for the juvenile European red-tooth bass shows that the SOD and CAT activities of the juvenile European red-tooth bass under the irradiation of red light are obviously higher than those of the treatment groups of green, white, yellow and blue light colors; performing 5 kinds of photochromic treatment on young plectropomus leopardus in the same gentle condition, and finding that SOD activity of a red light group sample is highest and AKP activity of a yellow light group sample is highest; gao et al also showed that the red group haliotis discus hannai has a higher SOD activity than the blue group.
In the description of the present invention, "a plurality" means two or more unless otherwise specified; the terms "upper", "lower", "left", "right", "inner", "outer", "front", "rear", "head", "tail", and the like, indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, are only for convenience in describing and simplifying the description, and do not indicate or imply that the device or element referred to must have a particular orientation, be constructed in a particular orientation, and be operated, and thus, should not be construed as limiting the invention. Furthermore, the terms "first," "second," "third," and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
The above description is only for the purpose of illustrating the present invention and the appended claims are not to be construed as limiting the scope of the invention, which is intended to cover all modifications, equivalents and improvements that are within the spirit and scope of the invention as defined by the appended claims.
Claims (4)
1. A macrobrachium rosenbergii larva breeding method is characterized by comprising the following steps:
and (3) salinity analysis: designing 16 per mill of high salinity group, 13 per mill of medium salinity group and 10 per mill of low salinity group; each group is provided with a plurality of parallels;
and (3) illumination analysis: all experimental barrels and internal light sources are covered by light-tight black mulching films, and LED bulbs with various colors are arranged right above the barrels; all power supply switching time is controlled by an automatic timer, and each group is provided with a plurality of parallels;
and (3) photoperiod analysis: designing 5 groups of experiment groups with different photoperiods, and irradiating by using white light, wherein the irradiation time is 0:24-h,6:18-h,12:12-h,18:6-h and 24: 0-h;
visible light analysis: designing different groups 5 of visible light experimental groups, wherein the visible light color wavelengths are red (615 nm-650 nm), yellow (580 nm-595 nm), white (450 nm-465 nm), green (495 nm-530 nm) and blue (450 nm-480 nm); the illumination period is 12: 12-h.
2. The macrobrachium rosenbergii larvae farming method according to claim 1, wherein in said salinity analysis 13% o of medium salinity groups are used, 3 parallel groups being set.
3. The macrobrachium rosenbergii larva breeding method of claim 1, wherein in the illumination analysis, all experimental barrels and internal light sources are covered by opaque black agricultural mulching films, and the light sources are 35W LED bulbs with various colors and are arranged at a position 30cm above the barrels;
each group is provided with 3 parallels; the salinity of water for each cultivation is 13 per mill.
4. The method for culturing the larvae of Macrobrachium rosenbergii according to claim 1, wherein the salinity analysis, the illumination analysis, the photoperiod analysis and the visible light analysis are carried out by:
and (3) data analysis, namely, the experimental data are sorted and analyzed, the data are processed by single-factor variance analysis, the LSD method mean value is subjected to multiple comparison, the P <0.05 shows that the significant difference exists, and the data are shown as the mean value +/-standard deviation (mean +/-SD).
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