CN108157297B - A kind of simple and efficient method for cultivating squama large bamboo hat with a conical crown and broad brim kentrogon - Google Patents
A kind of simple and efficient method for cultivating squama large bamboo hat with a conical crown and broad brim kentrogon Download PDFInfo
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- CN108157297B CN108157297B CN201711330730.8A CN201711330730A CN108157297B CN 108157297 B CN108157297 B CN 108157297B CN 201711330730 A CN201711330730 A CN 201711330730A CN 108157297 B CN108157297 B CN 108157297B
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- 235000017166 Bambusa arundinacea Nutrition 0.000 title claims abstract description 40
- 235000017491 Bambusa tulda Nutrition 0.000 title claims abstract description 40
- 241001330002 Bambuseae Species 0.000 title claims abstract description 40
- 235000015334 Phyllostachys viridis Nutrition 0.000 title claims abstract description 40
- 239000011425 bamboo Substances 0.000 title claims abstract description 40
- 238000000034 method Methods 0.000 title claims abstract description 25
- 241000195493 Cryptophyta Species 0.000 claims abstract description 22
- 241000238586 Cirripedia Species 0.000 claims abstract description 20
- 241000206751 Chrysophyceae Species 0.000 claims abstract description 7
- 230000012447 hatching Effects 0.000 claims abstract description 3
- 239000013535 sea water Substances 0.000 claims description 17
- 239000001963 growth medium Substances 0.000 claims description 13
- 238000005286 illumination Methods 0.000 claims description 9
- 241000975557 Dicrateria Species 0.000 claims description 5
- 241000196316 Tetraselmis subcordiformis Species 0.000 claims description 5
- 241000975561 Dicrateria inornata Species 0.000 claims description 2
- 238000011161 development Methods 0.000 abstract description 18
- 230000018109 developmental process Effects 0.000 abstract description 18
- 238000011534 incubation Methods 0.000 abstract description 10
- 238000002474 experimental method Methods 0.000 abstract description 8
- 238000011160 research Methods 0.000 abstract description 7
- 231100000463 ecotoxicology Toxicity 0.000 abstract description 4
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 6
- 238000012360 testing method Methods 0.000 description 5
- 241000206732 Skeletonema costatum Species 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 238000001035 drying Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 239000002574 poison Substances 0.000 description 3
- 231100000614 poison Toxicity 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 239000004576 sand Substances 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 238000012549 training Methods 0.000 description 3
- 241000345998 Calamus manan Species 0.000 description 2
- 241001501873 Isochrysis galbana Species 0.000 description 2
- 239000002519 antifouling agent Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 235000012950 rattan cane Nutrition 0.000 description 2
- 241000238421 Arthropoda Species 0.000 description 1
- 241001523707 Balanidae Species 0.000 description 1
- 241000238588 Balanus Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000206733 Skeletonema Species 0.000 description 1
- 241000351389 Tetraclita squamosa squamosa Species 0.000 description 1
- 241000196321 Tetraselmis Species 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000029052 metamorphosis Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000005868 ontogenesis Effects 0.000 description 1
- 230000010071 organism adhesion Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 239000011435 rock Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
Classifications
-
- 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
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of simple and efficient methods for cultivating squama large bamboo hat with a conical crown and broad brim kentrogon.After the egg hatching of squama large bamboo hat with a conical crown and broad brim barnacle is naupiar larva, launches flat algae and chrysophyceae is cultivated as bait.Squama large bamboo hat with a conical crown and broad brim barnacle naupiar larva rate of development can be improved in method of the invention, shortens incubation time, simplifies experimental procedure, to provide convenience with ecotoxicology research for ocean basis is ecological, application prospect is extensive.
Description
Technical field:
The invention belongs to field of biotechnology, and in particular to a kind of simple and efficient method for cultivating squama large bamboo hat with a conical crown and broad brim kentrogon.
Background technique:
Squama large bamboo hat with a conical crown and broad brim barnacle (Tetraclita squamosa squamosa (Bruguiere, 1789)) belongs to Arthropoda first
Shell animal subphylum jaw foot guiding principle sheath first subclass cirrus infraclass encloses chest catalogue stockless mesh Balanomorpha large bamboo hat with a conical crown and broad brim barnacle Superfamily large bamboo hat with a conical crown and broad brim Balanidae large bamboo hat with a conical crown and broad brim rattan
Pot subfamily large bamboo hat with a conical crown and broad brim Balanus, is distributed widely in China's coastal areas of the East and South China Seas, inhabites in intertidal zone and subtidal zone, be often attached to rock
On stone, harbour or buoy, become the important member in marine fouling organism group.
The history of life of squama large bamboo hat with a conical crown and broad brim barnacle is made of swim life and two stages of fixed life, wherein deviating from from naupiar larva
It is the life stage that swims that egg membrane, which explores and select the period of suitable attaching substratum to cyprids, and chooses attachment from cyprids
Then start to seek fixed life after base, settlement and metamorphosis.
The morphing process of complexity is undergone in ontogeny and is had based on squama large bamboo hat with a conical crown and broad brim barnacle swims and adhere to two kinds of lifes
The mode living totally different stage, therefore its larva is not only to carry out the ecological suitable object with ecotoxicology research in ocean basis,
And carries out marine fouling organism adhesion mechanism and prevent and kill off the gedanken experiment material of research.
Currently, existing disclosed technology is shown, with Skeletonema Costatum (Skeletonema under 20 DEG C of environment
Costatum), four slit bamboo or chopped wood algaes (Tetraselmis sp.) and Isochrysis galbana (Isochrysis galbana) mixed algae are bait
Material, the development that squama large bamboo hat with a conical crown and broad brim barnacle naupiar larva needs at least 14 days could become cyprids, and incubation needs persistently to fill
Gas, an addition seawater of antibiotic and replacement in every 2 days.As can promote larvae development, and reduction incubation time, simplified culture step,
It not only can shorten experimental period, improve working efficiency, reduce the probability of potential risk generation during test, but also can expand
Using this larva as the research contents of test object and working range, so that squama large bamboo hat with a conical crown and broad brim kentrogon be enable preferably to become related science
The gedanken experiment object of research and technological development.However, so far, domestic and international prior art is showed no the report of this respect.
Summary of the invention:
The present invention in order to overcome the shortcomings in the prior art, provides the training of a kind of promotion larvae development, shortening incubation time
Support the simple and efficient method of squama large bamboo hat with a conical crown and broad brim kentrogon.
Above-mentioned purpose that the invention is realized by the following technical scheme:
It is a kind of cultivate squama large bamboo hat with a conical crown and broad brim kentrogon simple and efficient method, be to squama large bamboo hat with a conical crown and broad brim barnacle egg hatching be naupiar larva
Afterwards, it launches flat algae and chrysophyceae is cultivated as bait.
It is preferred that the flat algae is P latymonas subcordifomis (Platymonas subcordiformis), the chrysophyceae is Zhan
River Dicrateria inornata (Dicrateria zhanjiangensis).
It is preferred that the dispensing flat algae and chrysophyceae are according to P latymonas subcordifomis (Platymonas subcordiformis)
Number ratio with station-service power source (Dicrateria zhanjiangensis) is that 1:1 is launched.
It is preferred that each injected volume of the P latymonas subcordifomis (Platymonas subcordiformis) be 1.2 ×
105A cell/mL, each injected volume of the station-service power source (Dicrateria zhanjiangensis) is 1.2 ×
105A cell/mL.
The culture is to be placed in naupiar larva in the culture medium of bait to cultivate in a dark environment, during culture
Replacement has the culture medium of bait and carries out intermittent illumination.
The culture medium that replacement has bait during the culture is specially that the training for having bait is replaced once after culture 5 days
Support medium.
Bait, which is launched, during the culture only carries out when larva culture starts and culture medium is replaced.
The cultivation temperature of the culture is preferably 29~31 DEG C.
Described is placed in naupiar larva the larva culture density cultivated in a dark environment in the culture medium of bait
Preferably 1~2/mL.
The carry out intermittence illumination is specially every morning or is put under natural light at dusk, illumination 0.5~1 hour.
The length of specific light application time depends on the microalgae suspension situation as bait.
The culture medium is preferably seawater.
The seawater is preferably the seawater by filtering, scalding.
Compared with prior art, the present invention has following remarkable result: the present invention provides one for the art and finishes
The method of complete feasible simple and efficient culture squama large bamboo hat with a conical crown and broad brim barnacle cyprids, can be improved squama large bamboo hat with a conical crown and broad brim barnacle naupiar larva rate of development, contracts
Short incubation time, simplifies experimental procedure, and the blank being filled in this technical field can be development ecotoxicology research and sieve
Larva culture technique needed for selecting marine fouling organism anti-fouling agent to provide and test object, meet basic research and technology development process
Involved in larvae development and attachment test etc. requirements of one's work.
The present invention solve squama large bamboo hat with a conical crown and broad brim kentrogon development time is longer, cultural method is relatively complicated, incubation need it is dynamic
The problems such as with air charging system, reducing extraneous factor influences the chance interfered, and ensure that experiment work can be single and controllable
Experimental situation in go on smoothly, Ecological basis research, ecotoxicology, anti-fouling agent screening and coating test etc. fields have
Wide application prospect.
Specific embodiment:
Technical solution of the present invention is further illustrated below by way of specific embodiment, rather than limiting the invention.
The embodiment of the present invention is since the performance of experimental facilities is limited, and 30 DEG C of constant temperature are fluctuated at 29~31 DEG C, and room temperature is 24~26
DEG C fluctuation, but without departing from above range.
Embodiment 1
By kentrogon cultivate vessel used cleaned, high-temperature sterilization and drying；Seawater then needs by sand filter, boils and disappear
Poison and etc. processing after, be cooled to room temperature spare.
The mature individual of squama large bamboo hat with a conical crown and broad brim barnacle then picks up from the reef of Daya Bay bank, picking individual as far as possible in collection process
Larger and lossless outwardly sound individual.Adult barnacle is dissected, the mature oosperm block of its brown is acquired, is put into equipped with 150mL
It filters in the 200mL transparent sample bottle of disinfected sea water and takes back laboratory, stood overnight in room temperature (about 24~26 DEG C) environment.
After pieces of an egg are largely hatched, collects active healthy and strong naupiar larva and is cultivated according to the following steps:
(1) using 1000mL glass beaker as culture vessel, appropriate filtering disinfected sea water and bait are added in beaker
The naupiar larva of active stalwartness is transferred in beaker by (about 1000mL), and larva culture density is about 2/mL.
(2) it places the beaker in 30 DEG C of constant incubators, is cultivated in dark condition, every morning (or at dusk) take out and burn
Cup, is put under natural light, illumination 1 hour, and the bait (microalgae) sunk to the bottom is made to float.
(3) larva culture is divided into three groups, respectively with P latymonas subcordifomis (flat algae bait group), station-service power source (gold
Algae bait group) and the algae solution that is mixed using number ratio 1:1 ratio of P latymonas subcordifomis and station-service power source as bait (mixed feed
Group), wherein the injected volume of single algae food is each about 2.4 × 105A cell/mL, and the injected volume of mixed feed is then sub-
Heart-shaped flat algae and station-service power source are respectively 1.2 × 105A cell/mL, the dispensing of bait only starts in experiment and culture is situated between
Matter carries out when replacing.
(4) every kind of feeding patterns set three Duplicate Samples, take out 30 childrens at random respectively from the other beaker of each group within every 2 days
Worm observes its development condition under the microscope after being killed with formalin, and records the quantity of each phase larva.After culture 5 days
A seawater is replaced, and adds corresponding bait again.
Experimental result is as shown in table 1, the larva that bait is fed using P latymonas subcordifomis and station-service power source hybrid mode
Rate of development was significantly faster than that the group individually fed using P latymonas subcordifomis (or station-service power source), and in culture 10 days
Afterwards, that is, there are a large amount of cyprids.Therefore, bait is used to train in such a way that flat algae and chrysophyceae are mixed in number ratio 1:1 ratio
Squama large bamboo hat with a conical crown and broad brim kentrogon is supported, effect is ideal, and not only larvae development speed significantly improves, and shortens incubation time, and can also obtain
To a large amount of cyprids.
Each phase larva percentage of squama large bamboo hat with a conical crown and broad brim barnacle of each experimental group under the different incubation times of table 1
Embodiment 2
By kentrogon cultivate vessel used cleaned, high-temperature sterilization and drying；Seawater then needs by sand filter, boils and disappear
Poison and etc. processing after, be cooled to room temperature spare.
The mature individual of squama large bamboo hat with a conical crown and broad brim barnacle then picks up from the reef of Daya Bay bank, picking individual as far as possible in collection process
Larger and lossless outwardly sound individual.Adult barnacle is dissected, the mature oosperm block of its brown is acquired, is put into equipped with 150mL
It filters in the 200mL transparent sample bottle of disinfected sea water and takes back laboratory, stood in room temperature (about 24~26 DEG C) environment, to nothing
After section larva largely hatches, collects active healthy and strong individual and is cultivated according to the following steps:
(1) using 1000mL glass beaker as culture vessel, appropriate filtering disinfected sea water and bait are added in beaker
The naupiar larva of active stalwartness is transferred in beaker by (about 1000mL), and larva culture density is about 1/mL.
(2) algae solution mixed using P latymonas subcordifomis and station-service power source using number ratio 1:1 ratio is bait, the Central Asia heart
The injected volume of shape flat algae and station-service power source is 1.2 × 105A cell/mL, the dispensing of bait only start and train in experiment
It is carried out when supporting Medium Replacement.
(3) larva culture is divided into two groups, and wherein A group places the beaker in 20 DEG C of constant incubators, and B group sets beaker
In in 30 DEG C of constant incubators, cultivated in dark condition, every morning (or at dusk) beaker is taken out, it is put under natural light, illumination
0.5~1 hour, the bait sunk to the bottom is made to float.
(4) every kind of training method sets three Duplicate Samples, takes out 30 childrens at random respectively from the other beaker of each group within every 2 days
Worm observes its development condition under the microscope after being killed with formalin, and records the quantity of each phase larva.After culture 5 days
A seawater is replaced, and adds corresponding bait again.
Experimental result is as shown in table 2, and the larvae development speed of B group (cultivation temperature is 30 DEG C) is significantly faster than that A group (culture temperature
Degree is 20 DEG C), and after culture 10 days, that is, there are a large amount of cyprids.Therefore, using 30 DEG C of temperature culture squama large bamboo hat with a conical crown and broad brim rattan
Pot larva, effect is ideal, and not only larvae development speed is apparently higher than 20 DEG C, shortens incubation time, and can also obtain
A large amount of cyprids.
Each phase larva percentage of squama large bamboo hat with a conical crown and broad brim barnacle of each experimental group under the different cultivation temperatures of table 2
Embodiment 3
By kentrogon cultivate vessel used cleaned, high-temperature sterilization and drying；Seawater then needs by sand filter, boils and disappear
Poison and etc. processing after, be cooled to room temperature spare.
The mature individual of squama large bamboo hat with a conical crown and broad brim barnacle then picks up from the reef of Daya Bay bank, picking individual as far as possible in collection process
Larger and lossless outwardly sound individual.Adult barnacle is dissected, the mature oosperm block of its brown is acquired, is put into equipped with 150mL
It filters in the 200mL transparent sample bottle of disinfected sea water and takes back laboratory, stood in room temperature (about 24~26 DEG C) environment, to nothing
After section larva largely hatches, collects active healthy and strong individual and is cultivated according to the following steps:
(1) using 1000mL glass beaker as culture vessel, appropriate filtering disinfected sea water and bait are added in beaker
The naupiar larva of active stalwartness is transferred in beaker by (about 1000mL), and larva culture density is about 2/mL.
(2) it places the beaker in 20 DEG C of constant incubators, is cultivated in dark condition, every morning (or at dusk) take out beaker
It is put under natural light, illumination 1 hour, the bait (microalgae) sunk to the bottom is made to float.
(3) larva culture is divided into 2 groups, wherein A group with Skeletonema Costatum, P latymonas subcordifomis and station-service power source with
The algae solution of number ratio 1:1:1 ratio mixing is bait, the i.e. injected volume of Skeletonema Costatum, P latymonas subcordifomis and station-service power source
It is 0.8 × 105A cell/mL.The algae solution that B group is mixed with P latymonas subcordifomis and station-service power source with number ratio 1:1 ratio
For bait, i.e. the injected volume of P latymonas subcordifomis and station-service power source is 1.2 × 105A cell/mL.The dispensing of bait only exists
It is carried out when experiment starts and culture medium is replaced.
(4) every kind of feeding patterns set three Duplicate Samples, take out 30 childrens at random respectively from the other beaker of each group within every 2 days
Worm observes its development condition under the microscope after being killed with formalin, and records the quantity of each phase larva.After culture 5 days
A seawater is replaced, and adds corresponding bait again.
As shown in table 3, the larvae development speed of A group is considerably slower than B group.Therefore, 20 DEG C at a temperature of, using sub- heart-shaped
The algae solution that flat algae and station-service power source are mixed using number ratio 1:1 ratio as bait culture squama large bamboo hat with a conical crown and broad brim kentrogon, effect be better than with
The algae solution that Skeletonema Costatum, P latymonas subcordifomis and station-service power source are mixed using number ratio 1:1:1 ratio is bait, larvae development
Speed is significantly improved, and shortens incubation time.
Each phase larva percentage of squama large bamboo hat with a conical crown and broad brim barnacle of each experimental group when 3 difference bait of table
Claims (8)
1. a kind of simple and efficient method for cultivating squama large bamboo hat with a conical crown and broad brim kentrogon, which is characterized in that the egg hatching to squama large bamboo hat with a conical crown and broad brim barnacle is no section
After larva, launches flat algae and chrysophyceae is cultivated as bait；The flat algae is P latymonas subcordifomis (Platymonas
Subcordiformis), the chrysophyceae is station-service power source (Dicrateria zhanjiangensis)；The Asia
Each injected volume of heart-shaped flat algae (Platymonas subcordiformis) is 1.2 × 105A cell/mL, described Zhanjiang
Each injected volume of Dicrateria inornata (Dicrateria zhanjiangensis) is 1.2 × 105A cell/mL.
2. the simple and efficient method of culture squama large bamboo hat with a conical crown and broad brim kentrogon according to claim 1, which is characterized in that the culture
It is to be placed in naupiar larva in the culture medium of bait to cultivate in a dark environment, replacement has the culture of bait to be situated between during culture
Matter simultaneously carries out intermittent illumination.
3. the simple and efficient method of culture squama large bamboo hat with a conical crown and broad brim kentrogon according to claim 2, which is characterized in that the culture
The culture medium that period replacement has bait is specially that the culture medium for having bait is replaced once after culture 5 days.
4. the simple and efficient method of culture squama large bamboo hat with a conical crown and broad brim kentrogon according to claim 2, which is characterized in that the culture
Period bait, which is launched, only to carry out when larva culture starts and culture medium is replaced.
5. the simple and efficient method of culture squama large bamboo hat with a conical crown and broad brim kentrogon according to claim 2, which is characterized in that the culture
Cultivation temperature be 29~31 DEG C.
6. the simple and efficient method of culture squama large bamboo hat with a conical crown and broad brim kentrogon according to claim 2, which is characterized in that described by nothing
It is 1~2/mL that section larva, which is placed in the larva culture density cultivated in a dark environment in the culture medium of bait,.
7. the simple and efficient method of culture squama large bamboo hat with a conical crown and broad brim kentrogon according to claim 2, which is characterized in that the progress
Intermittent illumination is specially every morning or is put under natural light at dusk, illumination 0.5~1 hour.
8. the simple and efficient method of culture squama large bamboo hat with a conical crown and broad brim kentrogon according to claim 2, which is characterized in that the culture
Medium is seawater.
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| CN101578967B (en) * | 2009-06-16 | 2012-07-25 | 中国科学院南海海洋研究所 | Method for culturing cyprids of balanus reticulates on large scale |
| CN102687708B (en) * | 2012-06-18 | 2013-10-16 | 中国船舶重工集团公司第七二五研究所 | Integrated control device for culture of barnacle larva |
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