CN112730168A - Method for measuring food intake of sedimentary feeding small benthonic animals - Google Patents
Method for measuring food intake of sedimentary feeding small benthonic animals Download PDFInfo
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Abstract
本发明公开了一种测定沉积食性小型底栖动物摄食量的方法，采用了基于水体颗粒有机碳含量测定的摄食量测定方法，由于沉积食性动物的食物以颗粒有机颗粒的形式被摄食，消化残渣排出体外后也成为悬浮或沉积颗粒有机物，因此根据其生活环境中颗粒有机碳的数量变化来计算某一时间段内的有机碳摄食量；在预先测定食物有机碳、蛋白质、脂肪、能量等营养成分含量的基础上，通过将有机碳摄食量乘以食物不同营养成分含量与有机碳含量的比值，从而换算为不同营养成分的摄食量。The invention discloses a method for measuring the food intake of sedimentary small benthic animals, which adopts a method for determining the food intake based on the determination of the content of particulate organic carbon in water bodies. After being excreted from the body, it also becomes suspended or deposited particulate organic matter, so the organic carbon intake in a certain period of time is calculated according to the change in the amount of particulate organic carbon in its living environment; On the basis of the component content, the food intake of different nutrients is converted into the food intake by multiplying the organic carbon intake by the ratio of the content of different nutrients in the food to the content of organic carbon.
Description
[ technical field ] A method for producing a semiconductor device
The invention relates to the technical field of animal food intake research, in particular to a method for measuring food intake of sedimentary feeding small benthonic animals.
[ background of the invention ]
Benthonic animals (zobenthos) refer to aquatic fauna living in the bottom of a water body for all or most of the life history, and are mostly fixed on hard matrixes such as rocks and buried in soft matrixes such as silt besides living in a fixed and active manner. In addition, there are also benthic species that attach to the surface of plants or other benthic animals, and that inhabit intertidal zones. On the feeding method, feeding in suspension and feeding in sediment are abundant. Most of the invertebrates are huge ecological groups. According to the size, large-scale benthonic animals and small-scale benthonic animals are divided, the species and the living mode of the animals are more complicated than those of zooplankton, and common benthonic animals comprise conch of gastropoda of mollusca, mussel of lamellibranchia, corbicula and the like; annelida, Lupinworm, etc. of Annelida, Whitmania pigra, etc. of Chodellodellida, etc. of Chodellida, Nereid of Chodellida, etc. of Nereid; midge larvae of entomophytes of the phylum arthropoda, dragonfly larvae, mayflies larvae and the like, shrimps, crabs and the like of the class crustacea; flat animal phylum, class of Whiteback Worm, and the like.
Most benthonic animals live in bottom sediment for a long time, have the characteristics of strong regionality, weak migration capacity and the like, generally have little avoidance capacity on environmental pollution and change, and relatively long time is needed for destroying and rebuilding communities; and most of the individuals are large and easy to identify; meanwhile, different species of benthonic animals have different adaptability to environmental conditions and tolerance and sensitivity degrees to adverse factors such as pollution and the like; according to the characteristics, parameters such as population structure, dominant species and quantity of the benthonic animals can be utilized to exactly reflect the quality condition of the water body.
The benthonic animals are classified into large-sized benthonic animals, small-sized benthonic animals and micro-sized benthonic animals according to their sizes. According to their life style, they are classified into a fixed type, a buried type, a drilled and eroded type, a benthic type and a freely movable type. The fixation type is animals living in water bottom or in water, such as marine sponge animals, coelenterates, tubal polychaetes, bryozoans, etc.; the bottom embedded type is animals which are embedded in water bottom mud to live, such as most hairy clams and clams of bivalve, crabs of cave, sea snake tails of echinoderm and the like; the bur-etch type is an animal living in wood stone, soil bank or stem and leaf of aquatic plant, such as sea bamboo shoot, ship maggot of mollusk and wood louse moth of crustacean; the benthic type is animals living on the surface of underwater soil and capable of moving slightly, such as gastropod mollusk, sea urchin, sea cucumber, starfish and other echinoderm animals; the freely movable type is an animal such as aquatic insect, shrimp, crab, etc. which crawls on the water bottom or swims in the water layer for a certain period of time. Most benthonic animals live in the bottom sediment for a long time, have the characteristics of strong regionality, weak migration capacity and the like, generally have little avoidance capacity on environmental pollution and change, and relatively long time is needed for destroying and rebuilding communities; most of the individuals are large and easy to identify; different species of benthonic animals have different adaptability to environmental conditions and tolerance and sensitivity degrees to adverse factors such as pollution and the like. According to the characteristics, parameters such as population structure, dominant species and quantity of the benthonic animals can be used for reflecting the quality condition of the water body.
At present, there are studies on biological food intake, for example, document CN201210274477 discloses a method for measuring individual food intake of fishes, and provides a method for measuring individual food intake of fishes aiming at the defects of radioactive hazard, inconvenient operation, difficult uniform mixing, large error, influence on fish food intake and the like in the prior art for measuring individual food intake of fishes, and the specific scheme is as follows: the metal particles with the diameter of about 0.25mm are screened and mixed with conventional feed according to a certain proportion to prepare feed containing the metal particles, then experimental fish is fed, the feed is absorbed by gastrointestinal tract and then the fish body is detected by an X-ray machine, and the food intake of the fish is indirectly measured according to the quantity of the metal particles. This document studies the food intake of individual fish and its conclusions do not necessarily apply to the study of food intake of small benthonic animals (surface and buried).
The food intake measurement of small benthonic animals (surface and buried) feeding on sediments has important significance in ecological research, and is one of research contents for providing basic data for the research of animal physiological ecology and the research of biogeochemical cycle of biogenic factors of habitat of the animals. Since the food source and fecal excretion of the predatory benthonic animals are on the surface of the sediment or inside the sediment, resulting in the mixing of food, sediment, and feces, it is difficult to separate and quantify them by a conventional separation method, and thus no effective method for measuring food intake has been developed. Although the identification and qualitative study of food sources and compositions of sedimentary food has been possible using biochemical and molecular biological methods, accurate quantification of food consumption has not been possible.
Therefore, there is a need to develop studies on food intake of sedimentary feeding small benthonic animals.
[ summary of the invention ]
Aiming at the problem that the food intake of small sedimentary feeding benthonic animals is difficult to measure in the existing ecological research, the invention provides a method for measuring the food intake of the sedimentary feeding small benthonic animals, which adopts a food intake measuring method based on the measurement of the organic carbon content of water body particles, because the food intake of the sedimentary feeding animals is fed in the form of the organic carbon particles, and the digestion residues are also suspended or deposited organic matters after being discharged out of the body, the organic carbon food intake in a certain time period is calculated according to the quantity change of the organic carbon particles in the living environment; the method is characterized in that the food intake is converted into the intake of different nutritional ingredients by multiplying the organic carbon intake by the ratio of the content of different nutritional ingredients of the food to the content of organic carbon after the content of the nutritional ingredients such as organic carbon, protein, fat, energy and the like of the food is measured in advance.
A method for determining food intake of sedimentary feeding small benthonic animals comprising the steps of:
1) the culture device comprises: according to the habit of the benthonic animals, quartz sand with a proper grain size is selected as sediment in a glass container with a proper water body size, and quartz sand with a certain thickness is paved according to the activity habit of the benthonic animals to be used as a domestication and experiment device for the benthonic animals;
2) determination of organic carbon of emission particles of benthonic animals: putting benthonic animals with certain biomass into the culture device which is not put and cultured with the benthonic animals and is in the step 1), after 24 hours, turning the culture device upside down and uniformly mixing, then quantitatively measuring the water body in the culture device, taking quantitative quartz sand, measuring the content of organic carbon particles in the water body and the quartz sand, and calculating the amount of organic carbon particles discharged by the benthonic animals within 24 hours according to the increase of the organic carbon particles in the culture device;
3) determination of organic carbon of feeding particles of benthic animals: adding quantitative food into the culture device, uniformly mixing the food into a water body of the culture device, then putting benthonic animals with certain biomass, after 24 hours, reversing the culture device, uniformly mixing the food, then quantitatively measuring the water body in the culture device, taking quantitative quartz sand, measuring the content of organic carbon particles in the water body and the quartz sand, and calculating the organic carbon intake of the benthonic animals within 24 hours according to the reduction amount of the organic carbon particles in the culture device.
The invention relates to a method for measuring food intake of sedimentary feeding small benthonic animals, which comprises the following steps:
1) the culture device comprises: selecting a glass container with a proper size as a culture container for measurement according to the biomass and the life habit of the benthonic animals;
2) substrate: selecting quartz sand with proper particle size as a substrate according to the life habit of the benthonic animals to be measured, determining the thickness of the quartz sand paved in the container according to the drilling depth of the benthonic animals, wherein the thickness is 1.5 times of the drilling depth, and weighing the weight (W) of the added quartz sand;
3) food preparation: drying food to be fed at 70 deg.C to constant weight, pulverizing with electric pulverizer, sieving with 60 mesh sieve, and measuring organic carbon content (TOC) with TOC analyzerf) And determining food Protein (PRO)f) Fat (LIP)f) Energy (ENR)f) Calculating the ratio relation of the nutrient content and the organic carbon;
4) pretreatment of the glass fiber filter membrane: keeping a 0.45-micron glass fiber filter membrane at 800 ℃ for 10 hours by using a muffle furnace, removing organic carbon possibly carried by the filter membrane, cooling, and sealing and storing for later use;
5) domestication: putting benthonic animals with certain biomass into a breeding device, adding water with proper volume (V), and domesticating the benthonic animals for 10 days according to the designed feeding amount and feeding mode;
6) blank control: adding quantitative food (I, g) into the culture device according to the same feeding amount and feeding mode of the domestication in the previous step, uniformly mixing the quantitative food (I, g) into the water body of the culture device, and taking no benthonic animals as a blank control experiment which is repeated for 3-5 times; after 24 hours, the cultivation device is inverted and uniformly mixed, then 100ml of water in the cultivation device is measured, after filtration by a pretreated glass fiber filter membrane, the filter membrane is dried in an oven at 70 ℃ to constant weight, and the organic carbon content is measured by a TOC tester; taking a sample of 100-150 g of quartz sand as a substrate, drying the sample in an oven at 70 ℃ to constant weight, measuring the content of organic carbon by using a TOC (total organic carbon) tester, and calculating the change proportion of the total organic carbon of food in the culture device within 24 hours according to the change of organic carbon particles in the culture device to be used as a correction factor (f) for calculating the food intake;
7) organic carbon determination of fecal excretion particles: according to the domestication density, putting benthonic animals with certain biomass into a breeding device which is not bred with the benthonic animals, not feeding food, and setting 10-12 times of organic carbon determination experiments of excrement excretion particles for repetition; after 24 hours, the culture device is inverted and uniformly mixed, then 100ml of water in the device is measured, the water is filtered by a pretreated glass fiber filter membrane, the filter membrane is dried in an oven at 70 ℃ to constant weight, and the organic carbon content is measured by a TOC tester; taking a sample of 100-150 g of quartz sand as substrate, drying the sample in an oven at 70 ℃ to constant weight, measuring the content of organic carbon by using a TOC (total organic carbon) tester, and calculating the amount of organic carbon (F) discharged by benthonic animals within 24 hours according to the increase of organic carbon in particles in the devicePOC，g)；
8) And (3) measuring the food intake of the granular organic carbon: adding quantitative food (I, g) into a culture device according to the same feeding amount and feeding mode in the domesticating process, uniformly mixing the quantitative food (I, g) into a water body of the culture device, then adding benthonic animals with certain biomass (B, g), and setting 10-12 times of experiments for determining the food intake of the granular organic carbon; at 24 hoursThen, the cultivation device is inverted and mixed evenly, then 100ml of water in the cultivation device is measured, after filtration is carried out by a pretreated glass fiber filter membrane, the filter membrane is dried in an oven at 70 ℃ to constant weight, and the organic carbon content is measured by a TOC tester; taking a sample of 100-150 g of the quartz sand as the substrate, drying the sample in an oven at 70 ℃ to constant weight, and measuring the content of organic carbon by using a TOC (total organic carbon) tester; calculating the total organic carbon content (E) of the culture device at the end of the experiment according to the measured resultsPOCG) and calculating the feed rate of the granular organic carbon (C) of the benthonic animals in 24 hours according to the reduction amount of the granular organic carbon in the breeding devicePOC，g)；
9) And (3) data calculation:
calculation of correction factor (f)
f-total organic carbon/total organic carbon dosed food after 24 hours with the blank control device
② amount of organic carbon in fecal excrementPOCG) calculating
FPOCThe organic carbon content of water body particles plus the organic carbon content of sediments of the device for feeding food without feeding food
(iii) calculating the total organic carbon content after experiment
EPOCExperimental device water body particle organic carbon content + sediment organic carbon content
(iv) food intake of granular organic carbon (C)POCCalculation of g)
CPOC＝I×TOCf-(EPOC-FPOC)/f
Quantitative calculation of nutrient intake (based on energy intake (C)EnergyJ) as an example)
CEnergy＝CPOC×(ENRf/TOCf)
In the formula CEnergy、ENRf、TOCfRespectively the energy intake, the food energy content and the food organic carbon content.
In the invention:
the glass container with the proper size in the step 1) is a glass container with the specification of 50 multiplied by 40cm3Aquarium or glass tank.
The quartz sand with the proper particle size in the step 2) is quartz sand with the particle size of 0.10-0.25 mm.
The thickness of the quartz sand in the step 2) is 10 cm.
The biomass in the step 5) is determined by placing 15 seedlings of Sipunculus nudus with the average weight of 0.5g into a container with the thickness of 50 multiplied by 40cm3The proportion in the aquarium.
Adding water with the proper volume in the step 5) is adding water with the volume equivalent to 80% of that of the culture device.
The biomass in step 6) is 50 multiplied by 40cm according to 15 seedlings cultured in Sipunculus nudus with the average weight of 0.5g3The proportion in the aquarium.
Compared with the prior art, the invention has the following advantages:
1. the method for measuring the food intake of the sedimentary feeding small benthonic animals can accurately calculate the food intake of the sedimentary feeding small benthonic animals.
2. The method for measuring the food intake of the sedimentary feeding small benthonic animals can calculate the intake of different nutrient components according to the ratio of the nutrient components of the bait to the content of organic carbon;
3. the method for measuring the food intake of the sedimentary feeding small benthonic animals can be applied to the food intake measurement of various small sedimentary feeding small benthonic animals.
[ detailed description ] embodiments
The following examples are provided to further illustrate the embodiments of the present invention.
Examples
A method for determining food intake of sedimentary feeding small benthonic animals comprising the steps of:
1 materials and methods
1.1 Gracilaria verrucosa offspring seeds
The sipunculus nudus seedlings for experiments are the sipunculus nudus seedlings bred by Guangxi ocean institute, are subjected to intermediate culture in an intermediate culture pond for about 4 months, are dug, caught and collected from the intermediate culture pond after the specification reaches about 0.5g/ind, and are selected for standby after 24 hours of clear water culture and sand spitting, wherein the seedlings have no damage to appearance, good activity and regular specification;
1.2 feed
The 4 feed raw materials for experiments are respectively feed for mangrove leaves, enteromorpha, cord grass and prawns; the red leaves are leaves collected on the beach, are taken back to a laboratory to be washed by clean water to remove silt, are washed by distilled water, are dried to constant weight at 70 ℃ after water control, are crushed by an electric crusher and are sieved by a 60-mesh sieve for later use; the Enteromorpha prolifera is collected on the beach, and the treatment method is the same as that of the mangrove leaves; the cord grass is a mixture of dead leaves and stems of the cord grass collected on mudflats, and the post-treatment method of the cord grass taken back to a laboratory is the same as that of the mangrove leaves; the prawn feed is prepared by mixing 5 commercially available prawn feeds in equal proportion, oven drying at 70 deg.C to constant weight, pulverizing with electric pulverizer, and sieving with 60 mesh sieve;
1.3 Experimental conditions
The experiment was conducted in an indoor workshop; the aquarium for cultivation has a specification of 50 × 40 × 40cm3Spreading sand 10cm at the bottom, and spreading water 10cm higher than the sediment; controlling temperature and feeding food for the experiment workshop, wherein the illumination is semitransparent shed roof transmission natural light, and the illumination period is consistent with the natural illumination period; the aquarium is continuously inflated with air; during the experiment, the salinity range of the seawater is 25-30, the pH value range is 8.0-8.3, the water temperature is 10 ℃ at the lowest in winter, and the water temperature is 32 ℃ at the highest in 7 months;
1.4 design of the experiment
1.4.1 acclimatization experiment
5 aquariums are used for each feed, 5 aquariums are arranged in parallel, and 20 aquariums are used for cultivation; putting 15 seedlings of Sipunculus nudus with the average weight of 0.5g into each aquarium, and weighing to obtain the initial weight (IW, g); different feed treatments are arranged in the aquarium by completely randomized treatment;
1.4.2 fecal excretion granule organic carbon and calibration factor determination
Putting benthonic animals with certain biomass into a conical flask in which the benthonic animals are not put according to domestication density, wherein the volume is 5L, quartz sand with the thickness of 10cm and the grain diameter of 0.10-0.25mm is paved at the bottom of the conical flask, and the organic carbon determination experiment of excrement and urine excretion grains is repeated for 10 times without feeding food; after 24 hours, the device is turned upside down and mixed, and then the device is measuredFiltering 100ml of internal water body by using a pretreated glass fiber filter membrane, drying the filter membrane in an oven at 70 ℃ to constant weight, and measuring the content of organic carbon by using a TOC (total organic carbon) tester; taking 100 g of a substrate quartz sand sample, drying the substrate quartz sand sample in an oven at 70 ℃ to constant weight, and measuring the content of organic carbon by using a TOC (total organic carbon) tester; calculating the amount of organic carbon (F) emitted from benthonic animals in 24 hours according to the increase of organic carbon in particles in the devicePOC，g)；
Adding quantitative food (I, g) according to the same feeding amount and feeding mode in the domesticating process in each experiment, uniformly mixing into the device water body, taking no benthonic animals as a control experiment, and repeating the control experiment for 3 times; after 24 hours, the device is turned upside down and mixed evenly, then 100ml of water in the device is measured, after filtration is carried out by a pretreated glass fiber filter membrane, the filter membrane is dried in an oven at 70 ℃ to constant weight, and the organic carbon content is measured by a TOC tester; taking 100 g of a substrate quartz sand sample, drying the substrate quartz sand sample in an oven at 70 ℃ to constant weight, and measuring the content of organic carbon by using a TOC (total organic carbon) tester; calculating the change proportion of the total organic carbon of the food in the device within 24 hours according to the change of the organic carbon of the particles in the device as a correction factor (f) for calculating the food intake;
1.4.3 food intake determination experiment
During the culture experiment period, the energy metabolism experiment is carried out once a month, and the experiment period is carried out for 7 times; measuring the food intake of Sipunculus nudus cultured with different feeds for 24h by using a sealed conical flask; the capacity of the conical flask is 5L, quartz sand with the particle size of 0.5mm and the thickness of 5cm is paved at the bottom of the conical flask, 4 Sipunculus nudus are put into each conical flask on the basis of a pre-experiment, 6 repeated reference flasks without the Sipunculus nudus are arranged for carrying out the experiment; firstly, putting 5g of feed into 50L of seawater, inflating to uniformly distribute the feed, then filling the feed into an experimental conical flask and a control conical flask, sealing the experimental conical flask and the control conical flask by using a preservative film, simultaneously taking 3 parts of water sample, filtering 3 parts of 100ml of water sample by using a 0.45 mu m glass fiber filter membrane, and storing the water sample by using the filter membrane to determine the TOC content; after 24 hours, reversing each conical flask for 15 times, uniformly mixing the water body, and respectively taking a water sample and quartz sand for treatment to determine the TOC content; the total amount of particulate organic carbon in the apparatus at the end of the experiment was calculated from the measurement results (E)POCG) and calculating the amount of suspended carbon in the device for 24 hours based on the amount of decrease in the particulate organic carbon in the deviceGranular organic carbon intake (C) of animalsPOC，g)；
1.5 sample composition determination
Drying food to be fed at 70 deg.C to constant weight, pulverizing with electric pulverizer, sieving with 60 mesh sieve, and measuring organic carbon content (TOC) with TOC measuring instrumentf) And determining food Protein (PRO)f) Fat (LIP)f) Energy (ENR)f) Calculating the ratio relation between the nutrient components and the organic carbon according to the content of the nutrient components; wherein TOC and TN are measured by an element analyzer, and energy content is measured by an oxygen bomb calorimeter; determining the TOC and TN contents of the filter membrane and the quartz sand by using an element analyzer;
1.6 data calculation
1.6.1 calculation of correction factor (f)
f-total organic carbon/total organic carbon dosed food after 24 hours with the blank control device
1.6.2 fecal Excretion particle organic carbon amount (F)POCG) calculating
FPOCThe organic carbon content of water body particles plus the organic carbon content of sediments of the device for feeding food without feeding food
1.6.3 Experimental facility Total organic carbon calculation
EPOCExperimental device water body particle organic carbon content + sediment organic carbon content
1.6.4 food intake of granular organic carbon (C)POCCalculation of mg)
CPOC＝I×TOCf-(EPOC-FPOC)/f
1.6.5 calculation of food intake (C, mg)
C＝CPOC/TOCf
1.6.6 quantitative calculation of nutrient intake
1.6.6.1 energy intake (C)Energy，J)
CEnergy＝CPOC×(ENRf/TOCf)
In the formula CEnergy、ENRf、TOCfEnergy intake, food energy content, and food organic carbon content, respectively;
1.6.6.2 food intake of crude protein (C)Protein，mg)
CEnergy＝CPOC×(PROf/TOCf)
PRO in the formulaf、TOCfRespectively food crude protein content and food organic carbon content, wherein PROfObtained by multiplying the total nitrogen content (TN) of the food by 6.25;
1.7 statistical analysis
Performing one-factor analysis of variance on data among different treatments, and performing Duncan's multiple comparison to check whether significant differences exist among different treatments; the percentage energy distribution data was arcsine transformed before analysis of variance
Statistical results in P<0.05 as a criterion for significant difference;
2 results of the experiment
2.1 survival and growth of Sipunculus nudus
The energy, organic carbon and nitrogen contents of the 4 baits are obviously different, and are shown in table 1;
TABLE 1 energy, organic carbon and nitrogen contents of four baits
Note: the numerical values in the same row without the same superscript letter are significantly different from each other;
2.2 food intake of Sipunculus nudus
The daily food intake of the Sipunculus nudus fed with 4 kinds of baits is shown in table 2, the organic carbon food intake is the highest based on prawn feed, the food intake weight and energy are the largest based on Enteromorpha prolifera, and the crude protein food intake is the largest based on prawn feed;
TABLE 2 organic carbon intake, feed weight, energy intake and crude protein intake of Sipunculus nudus on 4 feeds
Note: values in the same column that do not have the same superscript letter differ significantly from one another.
3, results:
the method for measuring the food intake of the sedimentary feeding small benthic animals can accurately calculate the food intake of the sipunculus nudus to organic carbon of different bait particles, and converts the food intake into food intake of different nutritional ingredients according to the ratio of the nutritional indexes of different baits to the content of the organic carbon, thereby providing great convenience for developing the physiological and ecological research of the sipunculus nudus food intake needing to be measured.
The above description is intended to describe in detail the preferred embodiments of the present invention, but the embodiments are not intended to limit the scope of the claims of the present invention, and all equivalent changes and modifications made within the technical spirit of the present invention should fall within the scope of the claims of the present invention.
Claims (6)
1. A method of determining food intake of sedimentary feeding small benthonic animals, comprising: the method specifically comprises the following steps:
1) the culture device comprises: selecting a glass container with a proper size as a culture container for measurement according to the biomass and the life habit of the benthonic animals;
2) substrate: selecting quartz sand with proper particle size as a substrate according to the life habit of the benthonic animals to be measured, determining the thickness of the quartz sand paved in the container according to the drilling depth of the benthonic animals, wherein the thickness is 1.5 times of the drilling depth, and weighing the weight (W) of the added quartz sand;
3) food preparation: drying food to be fed at 70 deg.C to constant weight, pulverizing with electric pulverizer, sieving with 60 mesh sieve, and measuring organic carbon content (TOC) with TOC analyzerf) And determining food Protein (PRO)f) Fat (LIP)f) Energy (ENR)f) Calculating the ratio relation of the nutrient content and the organic carbon;
4) pretreatment of the glass fiber filter membrane: keeping a 0.45-micron glass fiber filter membrane at 800 ℃ for 10 hours by using a muffle furnace, removing organic carbon possibly carried by the filter membrane, cooling, and sealing and storing for later use;
5) domestication: putting benthonic animals with certain biomass into a breeding device, adding water with proper volume (V), and domesticating the benthonic animals for 10 days according to the designed feeding amount and feeding mode;
6) blank control: adding quantitative food (I, g) into the culture device according to the same feeding amount and feeding mode of the domestication in the previous step, uniformly mixing the quantitative food (I, g) into the water body of the culture device, and taking no benthonic animals as a blank control experiment which is repeated for 3-5 times; after 24 hours, the cultivation device is inverted and uniformly mixed, then 100ml of water in the cultivation device is measured, after filtration by a pretreated glass fiber filter membrane, the filter membrane is dried in an oven at 70 ℃ to constant weight, and the organic carbon content is measured by a TOC tester; taking a sample of 100-150 g of quartz sand as a substrate, drying the sample in an oven at 70 ℃ to constant weight, measuring the content of organic carbon by using a TOC (total organic carbon) tester, and calculating the change proportion of the total organic carbon of food in the culture device within 24 hours according to the change of organic carbon particles in the culture device to be used as a correction factor (f) for calculating the food intake;
7) organic carbon determination of fecal excretion particles: according to the domestication density, putting benthonic animals with certain biomass into a breeding device which is not bred with the benthonic animals, not feeding food, and setting 10-12 times of organic carbon determination experiments of excrement excretion particles for repetition; after 24 hours, the culture device is inverted and uniformly mixed, then 100ml of water in the device is measured, the water is filtered by a pretreated glass fiber filter membrane, the filter membrane is dried in an oven at 70 ℃ to constant weight, and the organic carbon content is measured by a TOC tester; taking a sample of 100-150 g of quartz sand as substrate, drying the sample in an oven at 70 ℃ to constant weight, measuring the content of organic carbon by using a TOC (total organic carbon) tester, and calculating the amount of organic carbon (F) discharged by benthonic animals within 24 hours according to the increase of organic carbon in particles in the devicePOC，g)；
8) And (3) measuring the food intake of the granular organic carbon: in the culture device, the culture is performed according to the same domestication processAdding quantitative food (I, g) into the feeding amount and feeding mode, uniformly mixing the food and the feeding mode into a water body of a culture device, then putting benthonic animals with certain biomass (B, g), and repeating the particle organic carbon food intake measuring experiment by 10-12 times; after 24 hours, the cultivation device is inverted and uniformly mixed, then 100ml of water in the cultivation device is measured, after filtration by a pretreated glass fiber filter membrane, the filter membrane is dried in an oven at 70 ℃ to constant weight, and the organic carbon content is measured by a TOC tester; taking a sample of 100-150 g of the quartz sand as the substrate, drying the sample in an oven at 70 ℃ to constant weight, and measuring the content of organic carbon by using a TOC (total organic carbon) tester; calculating the total organic carbon content (E) of the culture device at the end of the experiment according to the measured resultsPOCG) and calculating the feed rate of the granular organic carbon (C) of the benthonic animals in 24 hours according to the reduction amount of the granular organic carbon in the breeding devicePOC，g)；
9) And (3) data calculation:
calculation of correction factor (f)
f-total organic carbon/total organic carbon dosed food after 24 hours with the blank control device
② amount of organic carbon in fecal excrementPOCG) calculating
FPOCThe organic carbon content of water body particles plus the organic carbon content of sediments of the device for feeding food without feeding food
(iii) calculating the total organic carbon content after experiment
EPOCExperimental device water body particle organic carbon content + sediment organic carbon content
(iv) food intake of granular organic carbon (C)POCCalculation of g)
CPOC＝I×TOCf-(EPOC-FPOC)/f
Quantitative calculation of nutrient intake (based on energy intake (C)EnergyJ) as an example)
CEnergy＝CPOC×(ENRf/TOCf)
In the formula CEnergy、ENRf、TOCfRespectively the energy intake, the food energy content and the food organic carbon content.
2. The method of claim 1 for determining food intake of sedimentary feeding small benthic animals, wherein: the glass container with the proper size in the step 1) is a glass container with the specification of 50 multiplied by 40cm3Aquarium or glass tank.
3. The method of claim 1 for determining food intake of sedimentary feeding small benthic animals, wherein: the quartz sand with the proper particle size in the step 2) is quartz sand with the particle size of 0.10-0.25 mm.
4. The method of claim 1 for determining food intake of sedimentary feeding small benthic animals, wherein: the thickness of the quartz sand in the step 2) is 10 cm.
5. The method of claim 1 for determining food intake of sedimentary feeding small benthic animals, wherein: the biomass in the steps 5) and 6) is determined by placing 15 seedlings of Sipunculus nudus with the average weight of 0.5g into 50 multiplied by 40cm3The proportion in the aquarium.
6. The method of claim 1 for determining food intake of sedimentary feeding small benthic animals, wherein: adding water with the proper volume in the step 5) is adding water with the volume equivalent to 80% of that of the culture device.
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