Systematic detection and mitigation of antimicrobial resistance in soil environment and animal health contributing to human health

The SiNorAMR project kick-off was on August 31 with partners from Norway (NIBIO, NMBU, NVI, Columbia Farms Ltd) and China (Nanjing Agricultural University, NJAU, and Heilongjiang Academy of Agricultural Sciences, HAAS) participating in the meeting. In addition, the Scientific Advisory Board members Nobel Laureate Sir Richard Roberts and Prof. Wei Dan, Beijing Academy of Agricultural and Forest Sciences, attended the kick-off meeting and delivered talks. The Research Council of Norway’s project officer was also present at the kick-off meeting and gave an opening speech followed by presentations of each work package (WP). The meeting was successful, and all partners understood well their role in the project and were motivated to contribute to the project. The Consortium Agreement was signed by all partners, a Data Management Plan was made and the project website was created (https://www.nibio.no/en/projects/sinoramr?locationfilter=true). During the first year, each WP has carried out the designated activities and worked interactively throughout the project. For WP1, project management and coordination, we have in addition to the above activities, organized four online and five physical meetings (one in China on July 2 in Harbin with NIBIO, NJAU and HAAS partners). Moreover, WP1 has distributed funding to the Norwegian and Chinese partners according to the contract and submitted the financial report for 2023. WP2 has had frequent communication with WP3 and WP4. The 160 DNA samples prepared by the NJAU partner (WP3) were shipped to WP2 at NIBIO Svanhovd for DNA chip development. The samples were evaluated, and a test run was conducted with some trouble-shooting work ongoing. The Norwegian Veterinary Institute has also collected samples which will be analysed. The WP3 led by NJAU has produced more results in four tasks (T1-4). T1: samples of animal intestines from piglets and finishing pigs of different breeds and disease status were collected for the characterization of ARB and ARGs. The abundances of ARGs and bacterial pathogens in four breeds of pigs, including Meishan pig, large Yorkshire, Min pig and Tibetan pig have been examined. The results show that: 1) breed affects the abundance of certain ARGs in pig intestines, E. coli is a common ARB in pig guts; 2) disease status significantly affects ARG abundances in pig guts. The abundances of several ARGs in the intestines of piglets with diarrhoea were significantly higher than those of healthy piglets. In T2, extracellular ARGs (eARGs) and intracellular ARGs (iARGs) have been characterized in commercial manure composts sampled across China. The results show that: 1) on average eARGs and iARGs account for 28.5% and 71.5% respectively of the total ARGs in composts, 2) plasmids play a crucial role in both the storage and horizontal transfer of eARGs, 3) compost eARGs and iARGs contain relatively high proportions of high-risk ARGs, and 4) 38 human pathogenic bacteria are identified as the hosts of eARGs and iARGs. This work is published in Science of the Total Environment (2024, 929: 172558). In T3, experiments were conducted to assess the impact of environmental factors, such as antibiotics, metals/metalloids and inner oxidative stress, on the transfer of plasmid-borne ARGs between bacteria. Results show that endogenous glutathione in bacteria plays an important role in restricting the conjugation of the ARG-carrying plasmid in pure culture and in soil. These results have been summarized in a manuscript for possible publication. In T4, incubation experiments have been conducted to study the effects of water management on soil resistome. Results show that: 1) antibiotic resistance determinants and the bacterial community respond differently to the key environmental factors, and 2) flooding could not accelerate the attenuation of soil ARGs or MGEs introduced from organic fertilizer but could prolong the half-life of MGEs in soil. WP4 led by the NMBU partner has established good collaboration with Columbia Farms Ltd and carries out the controlled pot experiments together with NMBU, NIBIO and Columbia Farms. Moreover, NMBU has hosted PhD student Wei Liu from NJAU during May-July 2024. The PhD student acquired training at NMBU and conducted the laboratory pilot experiments which showed a promising result with biochar, a recycled and waste-derived product, reducing the conjugation frequency of the conjugable R-plasmid pRAS1. Exp will be repeated. WP5 led by the 2nd NJAU partner has also started the designated task in advance (T1 designated for 2025) and initiated a pilot study on testing natural antibiotic substitutes Hesperidin× Rosmarinic acid on gut health and AMR of pig. A WP6 case study has been initiated by the HAAS and NIBIO partners. A detailed workplan is prepared. The 1st sample collections of manure from 60+ locations will be done in October 2024. Scientific publications and social media articles are produced in the 1st year and provided in the RCN form.

Antibiotics are widely used in animal husbandry to promote growth and prevent diseases, but global spread of antimicrobial resistance (AMR) and antimicrobial-resistant genes (ARGs) due to the abuse of antibiotics has caused severe challenge to "One Health" (www.who.int). Application of animal manures or manure-based organic fertilizers, especially from swine production which is a primary reservoir for antibiotic resistant bacteria (ARB), amplifies the risk of horizontal gene transfer (HGT) of AMR from animal production systems to the agricultural environment and food chain (e.g., vegetables and fruits), potentially impacting animal and human health. Our SiNorAMR project aims to tackle AMR by establishing a systematic detection method for AMR and ARGs to mitigate AMR in the soil agricultural ecosystem and control the ARGs and antimicrobial-resistant bacteria (ARB) during animal production through Norway-China collaboration, contributing to "One Health". The project is led by a senior female scientist with a consortium consisting of experienced senior group leaders balanced with junior scientists and PhD students from Norway (NIBIO, NMBU, NVI) and China (HAAS, NJAU) to assure successful execution of the project and to achieve the anticipated outcomes. Urban Gartneren AS in Norway play a key role in the designed two case studies. China ranks on the top in terms of antibiotic consumption and has imposed law enforcement to reduce the AMR problem. Through this SiNorAMR project, Norwegian experience in reducing and eliminating antibiotic use in animal feed and knowledge, methodologies and solutions developed in this project can be shared with our Chinese partners, and subsequently implemented in China in the future. Thus, this SiNorAMR project is a timely initiative to contribute to science-based solutions to support the policy and law enforcement to mitigate the AMR, a global threat to "One Health".