KR100777242B1 - Culture methods of a usefull ultra small rotifer, Synchaeta sp. as live food for hatch larvae with small mouth - Google Patents

Abstract

The present invention relates to a method for culturing a micro rotifer useful as a food organism of a small-sized hatching larva. More specifically, a micro rotifer having a size of 60 to 80 µm is subjected to a temperature condition of 16 to 24 ° C. and a salt condition of 10 to 15 ‰. Incubated with Tetraselmis suesica as a food organism, it is possible to grow hatchery fish, which have a small mouth and throat, which cannot be farmed by conventional feed organisms. It is characterized by culturing a culture solution containing an ultra-small rotifer having a former width of 37 to 50 µm with tetracelmis suecica as a food organism under a temperature condition of 16 to 24 ° C and a salt condition of 10 to 20 ‰.

Ultra-small rotator, cultivation method, hatching larvae, tetraselmis suesica

Description

Culture methods of a usefull ultra small rotifer, Synchaeta sp. as live food for hatch larvae with small mouth}

1 is an optical micrograph showing the morphological characteristics of the microscopic rotifers according to the present invention.

Figure 2 is an optical micrograph showing the size comparison between the microscopic yunchung and other yunchung according to the present invention.

Figure 3 is a growth curve graph according to the salinity condition of the micro lubricating insect according to the present invention.

Figure 4 is a graph of the growth curve according to the water temperature conditions of the micro lubricating insect according to the present invention.

Figure 5 is a growth curve graph according to the feeding conditions of the micro yunchung according to the present invention.

The present invention relates to a method for culturing a micro rotifer useful as a food organism of a small-sized hatching larva. More specifically, a micro rotifer having a size of 60 to 80 μm is subjected to a temperature condition of 16 to 24 ° C. and a salt condition of 10 to 20 ‰. By mass culturing with Tetraselmis suesica as a food organism, the small size of the mouth and throat is possible, so that hatchery can be cultured, which is impossible for normal food organisms.

At present, the main species of farming in our country is limited to flounder, skinball rock, rock bream, lobster, lobster. Moreover, since these species are mostly imported from China at low prices, domestic production is very disadvantageous in terms of price competitiveness, which may lead to the crisis of the domestic aquaculture industry.

In order to solve these problems, it is necessary to develop competitive high value-added target species. In this reality, in order to industrialize the development of high value-added target species, the planned supply and demand of seedlings must be successful. In addition to artificial breeding control technology, it is urgent to secure proper food quality and quantity. In other words, the effective production of such artificial seedlings requires large amounts of zooplankton to provide adequate food.

In general, there are many small zooplanktons in seawater, such as larvae of sponges, tonics and invertebrates, rotifers, and nauplius larvae of copepods. Plays an important role in the food chain. However, only a few of these zooplanktons have adequate conditions for feeding fish seedlings.

In recent years, many efforts have been made worldwide for the production of small-mouth fishes in the early stages of larvae. The main species of copepods, including the nautilus larvae and protozoa, are being studied. However, copepod napulis have to grow copepods of all stages in order to produce them, and the culture density is extremely low, and protozoa are promising such as Euprotes sp and Fabra salina. Although species were cultivated, they are not widely used for the production of seedlings as pathogenic or parasitic protozoa.

In addition, Brachionus plicatilis (large rotifers, 130-140 μm), a food organism that is absolutely used in aquaculture in Korea, and B. rotundiformis (B. rotundiformis), which are medium-sized species The rotifers of 100-210 µm) are also known as unsuitable foods for the viper species classified as high-quality fishes with small mouth and throat.

On the other hand, squids with small mouths and throats are tropical and subtropical species, which are distributed in the coast of Jeju, Korea, and because they are soft and delicious, they are the most expensive marine fish in the world's live fish market. The production of artificial seedlings is essential for farming, since yields are extremely limited.

However, in the reef fish, the mouth of the larvae waking from the egg is small and the throat is so narrow that it eats the larvae (large and small rotifers) that are used as food organisms in the production of common fish seedlings such as flounder, red snapper and stone bream. Seedling production is not successful.

Therefore, it is urgent to develop a zooplankton for a food organism that is small in size and easily cultivated for the production of fish whose mouth and throat are angled.

Therefore, an object of the present invention for solving the above problems of the above-mentioned conventional larvae feed organisms, Tetracell at 60 ~ 80㎛ size micro lubricating insect at 16 ~ 24 ℃ temperature conditions and 10 ~ 20 ‰ salt conditions By mass cultivation of Miss Sueshka as a food organism, it is possible to industrialize and stabilize artificial seedling production by developing micro-feeding organisms suitable for hatching larvae, which have been unable to grow with normal food organisms due to their small mouth and throat size. It is to provide a method of cultivating a micro rotifer useful as a food organism of a small hatched fish.

To achieve the above object, the microscopic yunchung useful as a food organism of the small hatched fish of the present invention,

Animal plankton to be used for hatching fish, characterized in that the armor is 60 ~ 80㎛ and the armor width is 37 ~ 50㎛.

In addition, the micro rotifer is characterized in that it was cultured with Tetraselmis suecica as a feed organism at a temperature condition of 16 ~ 24 ℃, salt conditions of 10 ~ 20 ‰.

And the method of cultivating a micro rotifer useful as a food organism of a small-mouth hatched fish,

Culture medium containing a micro rotator having a armor of 60 to 80 μm and a width of 37 to 50 μm was subjected to a temperature condition of 16 to 24 ° C. and a salt condition of 10 to 20 ‰,

It is characterized by culturing with Tetraselmis suesica as a feed organism.

In addition, it is characterized in that the use of isocrysis galvana as a supplementary feed of the micro rotator.

In addition, the incubation is characterized in that the temperature is made at 20 ℃ and salt conditions of 15 ‰.

Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

The ultra-small LSynchaeta sp. Of the present invention belongs to a large animal gate (Phylum Aschelminthes), Yunchunggang (輪 忠 綱, Class Rotifra), and has an armor of 60 to 80 µm and an armor of 37 to 50 µm. As plankton, the mouth and throat can be used as food for small hatcheries.

 The micro rotator of the present invention, which was collected in a lake area near the east coast, separated 60 to 80 μm microminiature larvae from the collected samples, and identified its morphological and ecological characteristics. The culture technique was developed.

As shown in FIG. 2, the ultra-small rotifers collected as described above are the smallest species of rotifers found so far with an individual size of 60 to 80 μm, and the armor as shown through the optical micrograph of FIG. 1. It is 60-80 micrometers, and the former width | variety is 37-50 micrometers, and it is a big possibility to use for food of a small mouth hatch.

Animal plankton of this size is suitable for the propagation of hatchery fish, especially among the hatchery fish such as red barley, larvae and gouper with small mouth and throat. Therefore, the micro rotator according to the present invention can be provided as a feed material for proliferation of hatch larvae of various seawater and freshwater fish, including squirrel fish, as a feed material for propagating hatched larvae. As a feed organism, it is preferable to incubate at a temperature condition of 16 to 24 ° C. and a salt condition of 10 to 20 ‰ as a feed organism, and more preferably to a temperature condition of 20 ° C. and a salt condition of 15 ‰. It is good to incubate in.

Detailed description of the culture conditions is made through the following culture method, the description thereof will be omitted.

Hereinafter will be described a method for culturing a micro rotifer according to the present invention.

First, the culture solution used in the present invention may be a liquid such as seawater, fresh water or brackish water, the type of the culture medium depends on the type of zooplankton to be cultured, depending on the type of zooplankton to be cultured The culture medium according to the present invention may be appropriately selected by those skilled in the art.

The culture conditions are not particularly limited, and suitable conditions may be selected by those skilled in the art according to the zooplankton to be cultured. However, the culture medium containing the micro lubricating insect according to the present invention has a temperature of 16-24 ° C. and a temperature of 10-20 ‰. It is preferable to be cultured in saline conditions, and more preferably, cultured at a temperature of 20 ° C. and a salt condition of 15 ‰ may exhibit excellent growth rate.

The food of the zooplankton used in the present invention may also be appropriately selected according to the zooplankton to be cultured as long as it does not affect the growth and survival of the zooplankton. Tetraselmis suecica having a cell size in the range of about 10 μm is most suitable, and isocrisis galvana may be used as a supplementary feed, and the amount of the feed may be determined according to the type of zooplankton to be cultured. It can be appropriately selected by.

The invention can be explained in detail through the following examples. However, the following examples are for illustrative purposes of the present invention and are not intended to limit the protection scope of the present invention.

(Example)

Collection and separation of micro rotifers were carried out as follows.

The ultra-small Synchaeta sp. Was collected and separated in the following manner.

Miniature Yunchung was collected from Gyeongpoho area, Gangneung-si. The collection point was a brackish point where seawater and freshwater meet. At that time, the salinity was 25 ‰ and the water temperature was 10 ℃. Collecting the animal plankton by using a plankton net of 50㎛ mesh, 30cm net, 1m in length was collected, and the collected sample was put in a 500ml water bottle and put in an ice box and transported to the laboratory. Samples transported to the laboratory were immediately taken out by capillary pipettes and housed in a single culture per one chamber of a zooplankton incubator (5ml x 12chamber). The initial culture water salinity was adjusted to 25 ‰ of the collection site, and the feed was diluted after dilution to 20,000 cells / ml of the small phytoplankton mixture of Isochrysis galbana and Tetraselmis suecica. It was. As the number of micro rotifers increased, they were housed in new chambers to secure species and to obtain samples for mass culture experiments.

The biological characteristics of the microminiature Synchaeta sp. Were as follows.

First, we looked at the morphological features of the tiny wheelworm.

Using microcapillary pipettes in a culture vessel, microscopic rotifers were collected one by one, placed on a slide glass, and morphological characteristics were observed under an optical microscope. The size was measured by dropping a drop of 5% neutral formalin on the slide glass to fix the sample, and then measuring the armor and the width of the micro rotator up to ± 1 μm with a ruler mounted on a microscope alternative lens.

As can be seen from the optical microscope picture 1, the ultra-small yunchung (Synchaeta sp.) Is in the shape of a screw and the armor and the range of 60 ~ 80㎛ and 37 ~ 50㎛, respectively, ultra-small yunhae (Synchaeta sp.) He found that he didn't have the skins that the existing rotifers had, but he had eye spots in the upper and middle parts of his body. It was able to perform swimming function, the upper middle part of the body has chewing machine (tooth) to perform the food intake function, and the lower part of the body was found to have claws (two).

In addition, we examined the ecological characteristics of the microscopic wheelworm.

The swimming speed of the micro rotator is similar to other rotifers, and the motion is linear rather than circular. However, if the environment was bad, it tended to sink in the circular motion in place and to be driven to a slightly darker place than the light.

In addition, the breeding method of micro rotifers appears differently depending on the environment, and it can be observed that the population increases through oviparum when the environment is good. In addition, an embryo (embryo) is formed in the female embryo area and gradually escapes to the bottom of the body as time passes, and the egg is attached to the foot area, and the embryo formation time and the embryo The time to get out of the body and turn into eggs varies depending on the environment. Freshly hatched micro rotifers in females carrying eggs do circular motions in place, but the range is not large, and as the time evolves, the range of movement becomes faster and wider. However, when the environment is not good, like the existing rotifers, the process of forming a durable egg could not be observed.

In order to examine the optimum culture conditions of the ultra-small Synchaeta sp., Each experiment was conducted under the following conditions.

 The microscopic rotifers collected and separated as described above were inoculated in a 250 ml Erlenmeyer flask (150 ml of cultivation solution) to carry out culture experiments with different salts, water temperature and feeding organisms.

Example 1

In order to investigate the salinity growth of the micro rotator, it was carried out as follows.

Salinity growth experiments of ultra-small Synchaeta sp. Were set to 5, 10, 15, 20, 25, and 30 ‰. The salinity of the culture solution was adjusted by mixing natural seawater (33 ‰) and freshwater (0 ‰) filtered through a filter, and inoculated at a density of 10 individuals / ml in a 250ml Erlenmeyer flask (150ml of culture solution) at 24 ° C. Incubated for 8 days in a maintained incubator. For feeding, Tetraselmis suecica was fed once a day at 8 × 10 ⁵ cells per 100 microscopic rotifers, and the whole was returned once every two days.

As shown in Table 1 and FIG. 3, the growth of the micro rotifers cultured in this manner was as shown in Tables 1 and 3, and the micro rotifers reached 118.0 individuals / ml on the 7th day in the 15 ‰ experimental group. The highest was 7.73%. After that, it was higher in order of 10, 20, 25 ‰. On the other hand, 5 and 30 ‰ were lower. As a result, the micro lubrication was found to be photo-inflammatory, and the growth tended to decrease as the salinity was higher and lower based on 15 ‰. Therefore, it is judged that 10 ~ 20 ‰ is the optimal culture salt for the micro rot, and 15 ‰, which has the highest density and the highest egg nesting rate, is considered as the most desirable salt concentration in the culture of this organism.

Growth of Miniature Synchaeta sp. According to Salinity salt(‰) Density (object / mL) Envelopment rate to the highest density (%) Individual Growth Rate (SGR) 5 77.0 ± 1.41 ^a 5.76 ± 0.34 ^a 0.60 ± 0.003 ^a 10 108.0 ± 3.77 ^ab 7.74 ± 0.38 ^b 0.65 ± 0.005 ^ab 15 118.0 ± 7.54 ^c 7.74 ± 0.12 ^b 0.67 ± 0.010 ^ab 20 105.3 ± 7.54 ^bc 6.91 ± 0.67 ^a 0.70 ± 0.060 ^b 25 96.3 ± 0.47 ^b 7.29 ± 1.27 ^a 0.64 ± 0.001 ^ab 30 77.1 ± 14.88 ^a 5.69 ± 0.07 ^a 0.60 ± 0.030 ^a

(The same superscript in each vertical line has no significant difference (95% confidence) as a result of statistical processing.)

Example 2

In order to examine the growth of the micro lubrication by water temperature was carried out as follows.

Ultra-small Synchaeta sp. Water growth zones were set to 16, 20, 24, 28 and 32 ℃. The incubator was adjusted to water temperature 16, 20, 24, 28 and 32 ° C. in a 250 ml Erlenmeyer flask (150 ml) with 15 ‰ culture water at a density of 5 individuals / ml and incubated for 10 days. . For feeding, Tetraselmis suecica was fed once a day at 8 × 10 ⁵ cells per 100 rotifers, and the whole was returned once every two days.

As shown in Table 2 and FIG. 4, the growth of the micro rotifers cultured in this way is shown in Table 2 and FIG. 4, and the same as the water temperature of 16, 20, and 24 ° C. was increased until the 5th day of culture, but the 6th day of culture. The highest density was found to be 190.3 individuals / ml on the 9th day of culture. In other experiments, the individual growth was low, and at 32 ° C., all died 2 days after the inoculation. Therefore, the micro rotifer can be cultured at a water temperature of 16 to 20 ° C. as a mesothermal species, and the optimum culture water temperature has been found to be 20 ° C.

Growth of Ultrasonic Synchaeta sp. According to Water Temperature Water temperature (℃) Density (object / mL) Envelopment rate to the highest density (%) Individual Growth Rate (SGR) 16 127.3 ± 3.75 ^b 9.14 ± 0.88 ^b 0.51 ± 0.036 ^c 20 190.3 ± 11.79 ^c 9.77 ± 1.38 ^b 0.62 ± 0.050 ^d 24 166.3 ± 7.31 ^b 10.05 ± 0.60 ^b 0.65 ± 0.058 ^d 28 14.3 ± 8.96 ^a 14.28 ± 0.29 ^c 0.27 ± 0.109 ^b 32 5.0 ± 0.00 ^a 0.00 ± 0.00 ^a 0.00 ± 0.000 ^a

(The same superscript in each vertical line has no significant difference (95% confidence) as a result of statistical processing.)

Example 3

To investigate the growth of micro rotifers by food, it was carried out as follows.

The foods used in the experiments of the micro-type synchaeta sp. Were Tetraselmis suecica (TET), Isochrysis galbana (ISO) and Chlorella ellipsoidea (CHL). It was paper, and the experiment was divided into three mixing spheres (TET + ISO, TET + CHL, ISO + CHL) and three single spheres (TET, ISO, CHL) for 13 days. In the experiment, the amount of feed was supplied once a day on the basis of 0.4 mg (3.0 × 10 ⁷ cells / ml) of dry weight per 1,000 individuals of CHL, TET, and ISO. Environmental conditions at the time of the experiment was the water temperature 20 ℃, 15 ‰ salinity and a total return once every two days.

As shown in Tables 3 and 5, the growth of the micro rotifers cultured in this way is shown in Table 3 and FIG. 5, and the highest density was 1566.7 individuals / ml on the 12th day of culture in the TET + CHL mixture. After that, the mixture of TET + ISO and TET alone were high without significant difference. On the other hand, ISO alone and ISO + CHL mixture showed low growth rate. In addition, in the case of the monocytes supplied with CHL, all died after the second day of culture. In other words, the experimental group did not contain tetraselmis suesica generally showed low growth.

Therefore, suitable for the cultivation of micro rotifers is tetraselmis suecica of the cell size of 10 ㎛ range is most preferred, Isochrysis galbana (Isochrysis galbana :) was determined to be used as a supplementary food.

Growth of Miniature Synchaeta sp. According to Feeding food Density (object / mL) Envelopment rate to the highest density (%) Individual Growth Rate (SGR) TET + ISO 991.1 ± 131.6 ^bc 18.97 ± 1.23 ^c 0.55 ± 0.020 ^c TET + CHL 1566.6 ± 464.8 ^c 18.63 ± 2.97 ^bc 0.59 ± 0.100 ^c ISO + CHL 33.4 ± 12.95 ^a 12.85 ± 2.82 ^b 0.23 ± 0.010 ^b TET ¹ 998.8 ± 272.7 ^bc 14.73 ± 2.74 ^bc 0.56 ± 0.020 ^c ISO ² 44.4 ± 6.0 ^a 15.65 ± 0.29 ^bc 0.25 ± 0.010 ^b CHL ³ 10.0 ± 0.0 ^a 0.00 ± 0.00 ^a 0.00 ± 0.000 ^a

(The same superscript in each vertical line has no significant difference (95% confidence) as a result of statistical processing.)

¹ TET: Tetraselmis suecica

² ISO: isochrysis galbana

³ CHL: Chlorella yirip Small Idea (Chlorella ellipsoidea)

As described above, the present invention has been described with reference to the above embodiments, but it is not necessarily limited thereto, and various modifications may be made without departing from the scope and spirit of the present invention.

As can be clearly seen from the above, according to the method of cultivating a micro rotifer useful as a feed organism of a small hatched larvae of the present invention, it is possible to isolate small animal plankton of 100 μm or less and to find the optimum culture conditions and to grow the mass culture. By inventing the technology, it is possible to multiply the optimum food organisms in a large amount under the optimal conditions, and it is possible to secure a large amount of food organisms in a short time with low production cost, thus greatly contributing to the aquaculture industry of high-quality fish species. Provides a useful effect.

In addition, by broadening the opportunity to produce small-mouthed fish after hatching, which is difficult to produce artificial seedlings in the past, domestic aquaculture technology can be increased by one step as fishery workers increase the production of new species, and fishery income It not only contributes greatly to the increase, but also provides the effect of resource recovery and resource creation of extinct marine fish species.

Claims (5)

delete delete Culture medium containing a micro rotator having a armor of 60 to 80 μm and a width of 37 to 50 μm was subjected to a temperature condition of 16 to 24 ° C. and a salt condition of 10 to 20 ‰, A method of culturing a micro rotifer useful as a feed organism for a small-mouth hatched larva, characterized by culturing Tetraselmis suesica as a feed organism. The method of claim 3, wherein A method for culturing a micro rotifer useful as a food organism of a small-sized hatching larva, characterized by using isocrysis galvana as an auxiliary food for the micro rotator. The method according to claim 3 or 4, The cultivation method of the micro yunchungsa useful as a food organism of small mouth hatching larva, characterized in that the temperature condition of 20 ℃ and salt condition of 15 ‰.

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