Method utilizing environmental sediment samples to monitor freshwater benthic animals

A benthic and sediment technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of cumbersome operation, high labor cost, long time consumption, etc., to ensure uniformity and overcome one-time collection. small amount of effect

Pending Publication Date: 2020-08-28
南京易基诺环保科技有限公司
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Problems solved by technology

[0002] Freshwater macrobenthos are one of the key monitoring objects for watershed ecological health assessment. The current monitoring process mainly consists of collecting sediment samples, manual picking, and morphological identification. High identification level and other limitations
By extracting the DNA in the sediment sample, the method of tracing the source of the sediment type through PCR amplification can effectively overcome the defects of long time-consuming, high labor cost and cumbersome operation in the prior art using morphological identification.

Method used

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  • Method utilizing environmental sediment samples to monitor freshwater benthic animals
  • Method utilizing environmental sediment samples to monitor freshwater benthic animals
  • Method utilizing environmental sediment samples to monitor freshwater benthic animals

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Embodiment 1

[0054] Sediment samples were collected from 72 sites in the Taihu Lake Basin in Jiangsu, including rivers, lakes, reservoirs and other water body types. The following steps will give a detailed introduction to the implementation method.

[0055] (1) Collection and preservation of environmental sediment samples

[0056] 1-1) Use tools such as a Peterson dredger and a triangular trawl to collect the bottom sediment of the water body. The bottom sediment contains plant residues, individual benthic animals and a small amount of sand and gravel.

[0057] 1-2) After filtering and rinsing the sediment through a screen, the mixture is basically drained of water, stored in a plastic bag, and frozen at -20°C to -80°C for 1-3 days.

[0058] (2) Pretreatment of environmental sediment samples

[0059] 2-1) Weigh the sediment sample after thawing, convert 1g of the mixture into 1ml, and add 3 to 5 times the volume of absolute ethanol to the sediment sample;

[0060] 2-2) After leaching with absolute ...

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Abstract

The invention belongs to the technical field of benthic animal identification, and discloses a method utilizing environmental sediment samples to monitor freshwater benthic animals. The method includes the following steps: 1) collecting an environmental sediment sample to preserve after cleaning; 2) adding anhydrous ethanol in the preserved environmental sediment sample to perform extraction treatment, taking uniformly mixed extract liquid after treating for a period of time, performing vacuum centrifugation on the extract liquid, and discarding supernatant to obtain dried tissue residues so as to perform DNA extraction; 3) utilizing the extracted DNA barcode fragment to perform amplification so as to obtain an amplified product; and 4) sequencing and analyzing the amplified product. The method performs extraction on the sample by adopting the anhydrous ethanol so as to make the DNA of a benthic animal uniformly released in the solution, so that a homogeneous state can be achieved; andthrough the collection of the ethanol solution containing the DNA sample and the extraction of the DNA after centrifugal concentration, the accurate monitoring of a wide range of environmental samples can be realized.

Description

Technical field [0001] The invention belongs to the technical field of benthic animal identification, and specifically relates to a method for monitoring freshwater benthic animals by using environmental sediment samples. Background technique [0002] Freshwater macrobenthos is one of the key monitoring objects of the ecological health assessment of the river basin. The current monitoring process is mainly collecting sediment samples, manual picking, and morphological identification. This process has a long time, high labor cost, and requires experts to Limitations such as high identification standards. By extracting DNA from sediment samples and tracing the types of sediments through PCR amplification, it can effectively overcome the defects of long time-consuming, high labor cost and cumbersome operation in the prior art using morphological identification. [0003] After searching, related applications have been disclosed in the prior art for the DNA extraction methods of organi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6869
CPCC12Q1/6869C12Q2531/113C12Q2565/125C12Q2535/122C12Q2537/165
Inventor 金珂杨江华张效伟
Owner 南京易基诺环保科技有限公司
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