CN111700011B - Temporary culture removal method for paralytic shellfish toxin - Google Patents

Temporary culture removal method for paralytic shellfish toxin Download PDF

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CN111700011B
CN111700011B CN202010332116.0A CN202010332116A CN111700011B CN 111700011 B CN111700011 B CN 111700011B CN 202010332116 A CN202010332116 A CN 202010332116A CN 111700011 B CN111700011 B CN 111700011B
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shellfish
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CN111700011A (en
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吴海燕
谭志军
张亚亚
张志华
郑关超
郭萌萌
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Hebei Aquatic Technology Promotion Station
Yellow Sea Fisheries Research Institute Chinese Academy of Fishery Sciences
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Yellow Sea Fisheries Research Institute Chinese Academy of Fishery Sciences
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; 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
    • A01K61/54—Culture of aquatic animals of shellfish of bivalves, e.g. oysters or mussels
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K63/00—Receptacles for live fish, e.g. aquaria; Terraria
    • A01K63/04—Arrangements for treating water specially adapted to receptacles for live fish
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/142—Amino acids; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23K20/00—Accessory food factors for animal feeding-stuffs
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    • A23K20/163—Sugars; Polysaccharides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
    • G—PHYSICS
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    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
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    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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    • G01N30/04—Preparation or injection of sample to be analysed
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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
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    • 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

Abstract

The invention discloses a temporary culture removal method of paralytic shellfish toxins, which comprises the following steps: (1) pretreatment of temporary shellfish culture; (2) treating a culture environment; (3) a rapid excretion stage; (4) a metabolism promoting stage; and (5) a fattening recovery stage. The invention adopts a systematic toxin elimination temporary rearing method to complete the stages of quick excretion, metabolism promotion and fattening recovery step by step. Compared with the conventional temporary rearing method, the method has the advantages that the adverse adaptation of the shellfish is promoted and harmful algae in the gastric contents are directly eliminated through the rapid excretion stage, so that the accumulation of toxins in other tissues and organs is reduced, and the purpose of shortening the temporary rearing period is achieved. The normal growth physiology of the shellfish in the temporary rearing stage is ensured by adjusting diet and enhancing immunity in different stages.

Description

Temporary culture removal method for paralytic shellfish toxin
Technical Field
The invention relates to a method for removing paralytic shellfish toxin by temporarily culturing scallop and mussel and application thereof. Belongs to the field of aquaculture.
Background
Paralytic shellfish toxins are marine biotoxins which are widely distributed and most harmful in the world and mainly comprise AlexandriumAlexandriumGymnodinium spGymnodiniumAnd dinoflagellate spPyrodiniumAnd isocellular dinoflagellate. The paralytic toxin harmful red tide frequently erupts offshore in China, alexandrium tamarense red tide occurs in the sea area near the south dry moat outside a city wall city of the Changtong island and the black reef sea area of 2012, PSTs exceed standards are detected in the Mytilus galloprovincialis of Qin and Huang islands in 2016, and a multi-person poisoning event is caused; the chain-shaped naked dinoflagellate has a plurality of red tide records in Bohai sea. In recent years, poisoning caused by eating bivalves occurs in China.
The scallop and the mussel have high water filtration rate, and can rapidly take harmful red tide in water in a short time, so that high concentration of pollutants such as paralytic shellfish toxin is accumulated, and quality safety risk is generated. In addition, researches prove that the toxigenic algae can release part of toxins into the water body during the harmful red tide, and the toxic algae can seriously affect the environment and shellfish. Therefore, under the same growth environment, the mussels and the scallops are varieties with high marine toxin accumulation content. The safety limit of the existing paralytic shellfish poison in China is 800 mug STX eq/kg, and the poisoning risk still exists in consumers. The reason is that the shellfish is complicated in variety and the yield is the first in the world, so that the supervision work is extremely difficult. Therefore, the efficient temporary shellfish culture method is an effective method for effectively discharging accumulated toxins in vivo, and the quality and safety of shellfish are guaranteed.
Common methods for removing toxins from marine organisms include physical and chemical methods. The temporary culture removal method is to feed aquatic product fed in polluted water area to the monitored area for a period of time to make the toxin in the body reach edible standard (CAC/RCP 52-2003: aquatic product and aquatic product processed product operating specification). In the temporary culture method at the present stage, the daily elimination rate of the quick and medium-speed toxin-expelling shellfish is about 15%, the highest elimination rate is about 63%, and the period is more than 21 days. It can be seen that the toxin eliminating effect of the existing temporary culture removing method is not obvious, and the method is not suitable for selling the shellfish with high yield and consumption. In the temporary culture stage, the autoimmune effect of the shellfish is obviously influenced by various indexes of the seawater including environmental requirements such as salinity, temperature, pH and the like and parameter requirements such as the type of fed baits, biomass and the like, so that the elimination of toxins and other pollutants is influenced.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: provides a technology for extracting, screening and mass spectrum confirmation of a plurality of esterified toxins of okadaic acid in algae cells.
The invention provides a method for removing paralytic shellfish toxins by temporary culture, which comprises the following steps:
(1) Pretreatment of temporary shellfish culture: washing shellfish with natural seawater to remove surface silt and shell, spreading and putting in temporary culture pond;
(2) Treatment of a culture environment:
the seawater for cultivation is prepared by blending deep-well seawater and seawater salt, wherein the salinity is required to be 25 to 30%; ventilating and standing for more than 12 to 24 hours;
the circulating water purification material is prepared by compounding filter cotton, fluorite and activated carbon material; wherein the activated carbon material is selected from 20 to 50 meshes, the weight ratio of the activated carbon material to the shellfish is 1.5g, the activated carbon material has the functions of purifying water quality and adsorbing water to remove toxins, and the activated carbon material sucks the bottom once every day in the morning;
(3) A rapid excretion stage: setting the water temperature to be 10 to 12 ℃; the method is characterized in that the dry spirulina powder and the fresh and alive bait algae are put in a matched mode; the feeding amount of the spirulina is 0.5 to 1.0g/m 3 The feeding amount of diatom is 10 7 ~10 8 Shellfish for 3-5 days;
(4) A metabolism promoting stage: gradually raising the water temperature to 12-13 ℃; feeding high-concentration fresh and alive bait algae of 10 9 Shell; simultaneously adding a compound immunopotentiator: taurine, peptidoglycan, chitosan oligosaccharide (molecular weight) and yeast, wherein the mass ratio of each component is 15 to 22:3~6:0.6 to 1.2:60 to 80 days for 7 to 10 days;
(5) A fattening recovery stage: the water temperature is set to be 13 to 15 ℃, and the feeding amount of the diatom is 10 7 ~10 8 Shellfish, and shellfish without paralysis until samplingToxoid, for 7-15 days.
Preferably, in the step (2), the salinity requirement in the culture environment treatment is 28%.
Preferably, in the step (2) of treatment of the culture environment, the activated carbon material is selected to be 50 meshes.
Preferably, in the step (3) rapid excretion stage, the water temperature is set to 10 ℃.
Preferably, in the step (4) metabolism promoting stage, the water temperature is raised to 13 ℃.
Preferably, the water temperature in the fattening recovery stage in the step (5) is set to be 15 ℃.
Preferably, the mass ratio of taurine, peptidoglycan, chitosan oligosaccharide (molecular weight) and yeast is 15:5:0.8:70.
preferably, the molecular weight of the chitosan oligosaccharide is 2.0-2.5kDa.
The invention has the beneficial effects that:
the invention ensures the quality safety of shellfish products, and temporarily cultivates the scallops and the mussels before the scallops and the mussels are sold in the market to remove paralytic shellfish toxins. By applying the method, the healthy development of the industry and the quality safety of consumers can be guaranteed, and meanwhile, the nutritional quality of scallop and mussel products can be improved. The invention adopts a systematic temporary culture method for toxin elimination, and finishes the stages of quick excretion, metabolism promotion and fattening recovery step by step. Compared with the conventional temporary rearing method, the method has the advantages that the adverse adaptation of the shellfish is promoted and harmful algae in the gastric contents are directly eliminated through the rapid excretion stage, so that the accumulation of toxins in other tissues and organs is reduced, and the purpose of shortening the temporary rearing period is achieved. The normal growth physiology of the shellfish in the temporary rearing stage is ensured by adjusting diet and enhancing immunity in different stages.
Drawings
The following describes embodiments of the present invention in further detail with reference to the accompanying drawings.
Fig. 1 is a tissue structure diagram of temporary rearing front mussels and temporary rearing back mussels.
Detailed Description
Example 1
The embodiment provides a method for providing temporary culture removal of paralytic shellfish toxins, which comprises the following steps:
(1) Pretreatment of temporary shellfish culture: washing shellfish with natural seawater to remove surface silt and remove empty shell, spreading and putting in a temporary culture pond;
(2) Treatment of a culture environment:
the seawater for cultivation is prepared by blending deep-well seawater with seawater salt, wherein the salinity is required to be 25 to 30%; ventilating and placing for more than 12 to 24 hours;
the circulating water purification material is prepared by compounding filter cotton, fluorite and active carbon material; the activated carbon material is selected from 20 to 50 meshes, the weight ratio of the shellfish is 1.5g, the activated carbon material has the effects of purifying water quality and adsorbing water to remove toxins, and the activated carbon material sucks the bottom once in the morning every day;
(3) A rapid excretion stage: in order to accelerate the rapid excretion of harmful red tide algae in stomach contents of scallops and mussels, the water temperature is set to be 10 to 12 ℃; the method comprises the following steps of (1) putting spirulina dry powder and fresh and alive bait algae (Chaetoceros minutissima) in a matching manner; the feeding amount of the spirulina is 0.5 to 1.0g/m 3 The feeding amount of diatom is 10 7 ~10 8 Shellfish for 5 days;
(4) A metabolism promoting stage: gradually raising the water temperature to 12-13 ℃. Feeding high-concentration fresh live bait algae (Chaetoceros minutissima) about 10 9 Shell only. Simultaneously adding a compound immunopotentiator: taurine, peptidoglycan, chitosan oligosaccharide (molecular weight) and yeast, wherein the mass ratio of the components is (15) - (22): 3~6:0.6 to 1.2:60 to 80 days for 7 days;
(5) And (3) fattening recovery stage: the water temperature is set to be 13 to 15 ℃, and the feeding amount of the diatom is 10 7 ~10 8 Shellfish for 10 days until no paralytic shellfish toxin is detected.
Example 2
Collecting natural perna canaliculus in the sea area of Qinhuang island in month 4, and detecting that the background content of paralytic shellfish poison is 1160 mu g STX eq/kg. The mussel shell has a length of 40.6 + -5.10 cm, a width of 22.4 + -3.50 cm, a weight of 6.84 + -1.05 g and a weight of 2.97 + -0.55 g, and has poor growth state and low meat yield of 69.7%.
(1) Pretreatment of temporary shellfish culture: washing shellfish with natural seawater to remove surface silt and shell, spreading and putting in temporary culture pond;
(2) Treatment of a culture environment:
the seawater for cultivation is prepared by blending deep well seawater and seawater salt, and has a salinity requirement of 28%; ventilating and standing for more than 24 hours;
the circulating water purification material is prepared by compounding filter cotton, fluorite and active carbon material; wherein 50g of the activated carbon material is selected to play a role in purifying water quality and adsorbing water to remove toxins, and the activated carbon material sucks the bottom once every morning;
(3) A rapid excretion stage: the water temperature is set to be 10 ℃; adopting a mode of putting spirulina dry powder and chaetoceros parviflora in a matching way; the feeding amount of spirulina is 0.5g/m 3 The feeding amount of diatom is 10 7 Shellfish for 5 days;
(4) A metabolism promoting stage: the water temperature was gradually increased to 13 ℃. Feeding Chaetoceros minutissima about 10 9 Shell only. Simultaneously adding 0.5g/L of compound immunopotentiator: taurine, peptidoglycan, chitosan oligosaccharide (molecular weight) and yeast, wherein the mass ratio of the components is 15:5:0.8:70, the time is 7 days; the molecular weight of the chitosan oligosaccharide is 2.0-2.5KDa
(5) And (3) fattening recovery stage: the water temperature is set to be 15 ℃, and the feeding amount of the diatom is 10 8 Shellfish for 10 days until no paralytic shellfish toxin is detected.
After the temporary rearing and removal for 22 days, the content of paralytic shellfish toxin in the mussels is detected to be 105 mu g STX eq/kg, and the mussels can be eaten safely. As a result, the results are shown in FIG. 1, which is a comparison of the histogram of mussels before temporary rearing (A) and after temporary rearing (B), wherein the mussel shell has a length of 48.4 + -3.10 cm, a width of 27.6 + -1.60 cm, a weight of 13.4 + -2.45 g, a weight of 3.55 + -0.94 g and a meat yield of 79.0%.
Example 3
Collecting Mytilus edulis in Qingdao sea area in month 4, and detecting to have no paralytic shellfish poison. The exposure test is carried out on mussels by using AT5-3 strain of Alexandrium tamarense which is typical paralytic shellfish poison toxigenic algae separated from sea areas of China. The feeding amount of the toxic algae is 8 multiplied by 10 6 cells/(∙ d only), after three days of continuous feeding, the content of paralytic toxin in the mussel is 1903 μ g STX eq/kg by detection.
(1) Pretreatment of temporary shellfish culture: washing shellfish with natural seawater to remove surface silt and remove empty shell, spreading and putting in a temporary culture pond;
(2) Treatment of a culture environment:
the seawater for cultivation is prepared by blending deep-well seawater with seawater salt, wherein the salinity is required to be 28 to 30%; ventilating and standing for more than 12 to 24 hours;
the circulating water purification material is prepared by compounding filter cotton, fluorite and active carbon material; wherein 50g of 50 meshes of activated carbon material is selected to play the roles of purifying water quality and adsorbing water to remove toxins, and the activated carbon material sucks the bottom once every morning;
(3) A rapid excretion stage: the water temperature is set to be 10 ℃; the method adopts the way that the spirulina dry powder and the chaetoceros gracilis are matched and put into the reactor; the feeding amount of spirulina is 0.5g/m 3 The feeding amount of diatom is 10 7 Shellfish for 5 days;
(4) A metabolism promoting stage: the water temperature was gradually increased to 13 ℃. Feeding Chaetoceros minutissima about 10 9 Shell is/only. Simultaneously adding 0.5g/L of compound immunopotentiator: taurine, peptidoglycan, chitosan oligosaccharide (molecular weight) and yeast, wherein the mass ratio of the components is 15:5:0.8:70, the time is 7 days;
(5) A fattening recovery stage: the water temperature is set to be 15 ℃, and the feeding amount of the diatom is 10 8 Shellfish for 10 days until no paralytic shellfish toxin is detected.
After the temporary rearing and removal for 22 days, the content of paralytic shellfish toxin in the mussels is detected to be 210 mu g STX eq/kg, and the mussels can be eaten safely.
The foregoing is directed to preferred embodiments of the present invention, other and further embodiments of the invention may be devised without departing from the basic scope thereof, and the scope thereof is determined by the claims that follow. However, any simple modification, equivalent change and modification of the above embodiments according to the technical essence of the present invention are within the protection scope of the technical solution of the present invention.

Claims (1)

1. A method for providing temporary culture removal of paralytic shellfish toxins, comprising the steps of:
(1) Pretreatment of temporary shellfish culture: washing shellfish with natural seawater to remove surface silt and remove empty shell, spreading and putting in a temporary culture pond;
(2) Treatment of a culture environment:
the seawater for cultivation is prepared by adding seawater salt into deep well seawater, and the salinity is required to be 25 to 30 per mill; ventilating and standing for 12 to 24 hours;
the circulating water purification material is prepared by compounding filter cotton, fluorite and active carbon material; wherein the active carbon material is selected from 20 to 50 meshes, plays the roles of purifying water quality and adsorbing water to remove toxins, and sucks the water bottom once every morning;
(3) A rapid excretion stage: in order to accelerate the rapid excretion of harmful red tide algae in stomach contents of scallops and mussels, the water temperature is set to be 10 to 12 ℃; adopting a mode of putting spirulina dry powder and chaetoceros parviflora in a matching way; the feeding amount of the spirulina is 0.5 to 1.0g/m 3 The feeding amount of the Chaetoceros minutissima is 10 7 ~10 8 Shellfish for 5 days;
(4) A metabolism promoting stage: gradually raising the water temperature to 12 to 13 ℃, and feeding high-concentration Chaetoceros parvulus into the water until the water temperature is 10 DEG 9 Shellfish, with the addition of a compounded immunopotentiator: taurine, peptidoglycan, chitosan oligosaccharide and yeast, wherein the mass ratio of the components is (15) - (22): 3~6:0.6 to 1.2:60 to 80 days for 7 days;
(5) And (3) fattening recovery stage: the water temperature is set to be 13 to 15 ℃, and the feeding amount of the diatom is 10 7 ~10 8 Shellfish for 10 days until no paralytic shellfish toxin is detected.
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