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CN-122024824-A - Microplastic pollution antibiotic resistance gene risk evaluation method based on relative and absolute abundance

CN122024824ACN 122024824 ACN122024824 ACN 122024824ACN-122024824-A

Abstract

The invention relates to the field of risk assessment of environmental Antibiotic Resistance Genes (ARGs), and provides a double-index collaborative risk assessment method for introducing Meta analysis, aiming at the problems that conclusion of microplastic on ARGs in the existing study is inconsistent, and evaluation is easy to be caused only by dependence on relative abundance (abundance normalized by taking 16S rRNA genes and the like as references) or absolute abundance (abundance based on copy numbers). The method comprises the steps of data acquisition, index processing, meta analysis and multi-factor adjustment, double index interpretation, risk assessment and the like. By integrating ARG relative abundance and absolute abundance data of a micro-plastic exposure group and a control group and combining factors such as environmental matrixes, exposure time, particle size, concentration, type and the like, the influence trend of micro-plastic on ARG abundance under different conditions is evaluated, risk types are judged from a double-index view, and accuracy and applicability of resistance risk evaluation are improved.

Inventors

  • YANG BAOSHAN
  • Huang Gengkui
  • WANG HUI

Assignees

  • 济南大学

Dates

Publication Date
20260512
Application Date
20260202

Claims (7)

  1. 1. A method for evaluating the risk of a micro plastic pollution antibiotic resistance gene based on relative and absolute abundance, which is characterized by comprising the following steps: s1, determining an evaluation index and a data collection range Setting a double index of the abundance of the resistance genes, namely the relative abundance and the absolute abundance, as a core index for evaluating the influence of the microplastic on the ARG abundance; S2, document retrieval Collecting researches related to the influence of micro plastic exposure on ARG abundance in a plurality of databases through keyword retrieval documents, screening the documents according to a priority report Project (PRISMA) of systematic evaluation and Meta analysis, rejecting the researches which do not meet the inclusion standard, and incorporating the researches meeting the conditions into a Meta analysis database; S3, data extraction Extracting basic information of articles, ARG abundance and related data of microplastic from the incorporation study, extracting numerical values of the data which are presented in a graph and cannot be directly read by a digitizing tool, and collecting all the extracted data for subsequent analysis; s4, data processing and effect calculation Processing the extracted data and calculating the effect quantity of each test comparison, wherein the effect quantity is expressed by natural logarithm (ln RR) of the average value ratio of a processing group to a comparison group, and corresponding sampling variance is calculated for weighted analysis; S5 Meta analytical modeling Constructing a multi-level random effect Meta analysis model on the effect quantity data set, and setting the identification of each independent study as a random effect factor to consider the correlation of multiple effect quantities in the same study; S6, heterogeneity analysis Calculating the Cochran's Q statistic and the significance level thereof to evaluate the heterogeneity of the total effect quantity, and determining whether a significant regulating effect exists or not through the difference of the effect quantity among different levels of each regulating variable of Q statistical test (QM) of Meta regression; s7, result visualization and publishing bias evaluation Drawing an orchard graph based on a Meta analysis result, visually presenting the effect quantity and 95% confidence intervals of each test comparison in a graphical mode, and simultaneously drawing a funnel graph and Egger's test to evaluate the robustness and the credibility of the Meta analysis result; S8, interpretation of results And analyzing effect results under two indexes of relative abundance and absolute abundance, and judging the risk type represented by ARG change caused by exposure of the microplastic.
  2. 2. The method of claim 1, wherein the literature search and screening in step S2 follows PRISMA system specifications, and the selected study is satisfied by a microplastic exposure experimental design with a set control group, provides an average value and standard error of the abundance of the resistance genes, has no less than 3 independent repetitions, and explicitly reports key information such as polymer type, particle size, dosing concentration, environmental substrate and the like of the microplastic.
  3. 3. The method of claim 1, wherein the step S4 of correcting the detection limit for the observation data with the standard error report of 0 comprises multiplying the minimum value by 0.7 as the estimated detection limit when the other resistance genes in the same test have the reported minimum average value, multiplying the minimum value of all the data of the category to which the genes belong by 1.2 as the estimated detection limit when the internal correlation data of the same test are absent, and multiplying the global minimum value of the observation values in the whole Meta analysis dataset by 0.8 as the detection limit if the basis is not obtained yet. And setting the corrected average value as half of the estimated detection limit, and setting the corrected standard deviation as one fourth of the estimated detection limit to replace the original observed data for effect quantity calculation.
  4. 4. The method of claim 1, wherein the effect in step S4 is expressed as natural logarithm of the ratio of the mean of the microplastic treated group to the mean of the control group, and the calculation formula is: wherein Is the average value of the microplastic treatment group, As an average value of the control group, And the sampling variance of the effect quantity is calculated according to a standard deviation of a processing group and a control group and a sample quantity according to a set formula, and the effect quantity ln RR can be converted into a corresponding percentage change rate for result interpretation.
  5. 5. The method according to claim 1, wherein the Meta analysis model in step S5 incorporates five factors of polymer type, microplastic particle size, microplastic concentration, exposure duration, and environmental matrix as classification adjustment variables into the model, and evaluates the effect of each factor level on the effect.
  6. 6. The method of claim 1, wherein step S7 further comprises publishing bias assessment by plotting a funnel of Meta analysis results and detecting the published bias using Egger' S regression test, if the funnel is significantly asymmetric, estimating potential missing studies using a Trim-and-Fill method (Trim-and-Fill) and adjusting the aggregate effect values to ensure robustness of the resulting conclusions.
  7. 7. The method according to claim 1, wherein the step S8 is characterized in that the risk of the propagation of the resistance gene related to the microplastic is judged by simultaneously analyzing the results of the two effects of the relative abundance and the absolute abundance, wherein the exposure of the microplastic is judged to lead to the actual amplification of the resistance gene and the increase of the risk of the environmental resistance only when the effect values of the two abundance indexes are obviously increased and are consistent, and the risk is judged to be possibly overestimated if the increase of the relative abundance index is detected only and the absolute abundance is not obviously increased or even decreased, and the situation that the deviation conclusion is reached based on a single index is required to be alerted in the risk assessment.

Description

Microplastic pollution antibiotic resistance gene risk evaluation method based on relative and absolute abundance Technical Field The invention belongs to the technical field of evaluation of environmental pollution risks of antibiotic resistance genes, and particularly relates to an evaluation method for evaluating influence of exposure of microplastic on ARG abundance. Background ARG can be transmitted between environmental microorganisms and human pathogens, impairing antibiotic efficacy, and has become a significant challenge for public health worldwide. Microplastic (MPs) widely exist in ecological systems such as water, soil and the like, have high specific surface area and stable physicochemical properties, can widely interact with microorganisms, organic pollutants, heavy metals and the like, rapidly form a plastic ring biomembrane, and become an active carrier for promoting microorganism enrichment and gene exchange. Microplastic may affect the abundance and diffusion of ARG in the environment by providing mechanisms to attach substrates and promote horizontal gene transfer, etc. However, the results of the prior literature on the effect of microplastic on ARG abundance are not consistent, some studies have found that microplastic exposure can promote a significant increase in ARG, others have observed conditions of insignificant effect or even a decrease in abundance. This conclusion difference reflects the lack of systematic quantitative analysis, which is detrimental to the overall knowledge of the problem. In addition, most researches are evaluated by only using one index of the relative abundance or absolute abundance of ARG, so that risk judgment is easy to be achieved on one side, and conditions such as environment substrates, exposure time and micro plastic characteristics of different researches are different, and the results are difficult to compare due to the lack of uniform integration. These deficiencies indicate that it is necessary to build a unified assessment framework. Meta analysis is a powerful tool to integrate multiple independent research results on the basis of standardized effectors, evaluate overall trends, and identify sources of heterogeneity. However, no global Meta analysis for the effect of microplastic exposure on ARG abundance is currently available, neither explicit comparison of differences in relative abundance versus absolute abundance index nor systematic study of the co-regulation of multiple factors such as environment and materials. Therefore, the development of a comprehensive evaluation method with double index cooperation and multi-factor regulation is of great significance for understanding the mechanism of microplastic driving resistance transmission in depth and improving risk evaluation. Disclosure of Invention The invention provides an evaluation method for the ARG abundance of a microplastic influence environment, and aims to solve the problems that in the prior art, risk judgment deviation is caused by single index analysis, heterogeneity of different research results is difficult to explain and the like. Aiming at the problems of scattered data, non-uniform indexes, scene dependence and the like in the research of the exposure of the micro-plastics, the invention provides a method for analyzing 'double index' multi-dimensional Meta, which combines the relative abundance with the absolute abundance to carry out comprehensive analysis so as to distinguish the composition change of a microbial community from the total ARG change, thereby more accurately evaluating the antibiotic resistance risk related to the micro-plastics. The technical scheme of the method comprises the following steps: s1, determining an evaluation index and a data collection range Setting a double index of the abundance of the resistance genes, namely the relative abundance and the absolute abundance, as a core index for evaluating the influence of the microplastic on the ARG abundance; S2, document retrieval Collecting researches related to the influence of the exposure of the microplastic on ARG abundance in a plurality of databases through keyword search documents, screening the documents according to PRISMA, eliminating the researches which do not meet inclusion criteria, and incorporating the researches meeting the conditions into a Meta analysis database; S3, data extraction Extracting basic information of articles, ARG abundance and related data of microplastic from the incorporation study, extracting numerical values of the data which are presented in a graph and cannot be directly read by a digitizing tool, and collecting all the extracted data for subsequent analysis; s4, data processing and effect calculation Processing the extracted data and calculating the effect quantity of each test comparison, wherein the effect quantity is expressed by natural logarithm (ln RR) of the average value ratio of a processing group to a comparison group, and corresponding sampling var