The WHO considers that antimicrobial resistance (AMR) is among the ten greatest global public health risks of the 21st century. The expansion of human populations and anthropogenically related activities, accompanied by the fragmentation of natural habitats, has resulted in increased human–wildlife interaction. Natural ecosystems are therefore subjected to anthropogenic inputs, which affect the resistome of wild animals. Thus, urgent multisectoral action is needed to achieve the Sustainable Development Goals following the One Health approach. The present work falls within the scope of this approach and aims to characterize the AMR of the faecal microbiome of the red fox (Vulpes vulpes), an opportunistic and generalist synanthropic species whose abundance has been increasing in urban and peri-urban areas. A high number of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) were screened and quantified using a high-throughput qPCR approach, and the antimicrobial susceptibility of cultivable E. coli and Enterococcus spp. were assessed interpreted with both ECOFFs and clinical breakpoints. The most abundant ARGs detected confer resistance to trimethoprim and tetracyclines, although the first were absent in one of the locations studied. Several ARGs considered to be threats to human health were identified in high relative abundances (bla_TEM, ermB, aadA, tetM, tetW, tetL, drfA1 and drfA17), especially in the geographical area with greater anthropogenic influence. Although at a low percentage, resistant and multidrug-resistant (MDR) E. coli and Enterococcus spp. were isolated, including one MDR E. coli showing resistance to 12 antimicrobials from 6 different classes.

In the last decades, the wildlife-human interface has been increasing due to several anthropogenic factors. Therefore, it is crucial to be aware of the impact of these new dynamics on the health of wild animals and its associated zoonotic disease risks. This study aimed to characterise the faecal microbiota of two populations of red deer (Cervus elaphus) by metabarcoding, with a particular focus on potential human and veterinary pathogens, and to perform an assessment of antibiotic resistance genes (ARGs) ocorrence. The faecal microbiota of red deer was assessed by metabarcoding using the 16S rRNA marker, and OTUs of the genera Treponema, Yersinia, Clostridium, Mycobacterium and Rickettsia were identified. Two of them affiliated with species more commonly regarded as pathogens (Clostridium piliforme and Yersinia enterocolitica). The quantification of ARGs was performed by quantitative real-time PCR, using a metagenomic approach, and the most abundant genes were found to be bla_TEM, sul1, tetracycline resistance genes (tetW, tetO and tetQ) and ermF. From these, tetO and tetW are rank II ARGs, which were recently considered future threats for human health. Our results suggest the need for screening programs for the occurrence of pathogens and ARGs in wildlife and particularly in game species. This article is protected by copyright. All rights reserved

Antimicrobial resistance (AMR) is a global health concern. Nowadays, antibiotic resistance genes (ARGs) are considered emerging pollutants. This study followed the One Health framework, in which AMR surveillance in the environment, including in wild animals, is advisable to mitigate this problem. Here we investigated AMR associated with Eurasian otter, a semi-aquatic mammal considered an indicator of freshwater health. To do so, otter's faecal resistome was characterized by a high-throughput qPCR array. This technique has a high-capacity of ARGs profiling. Additionally, we have assessed the antimicrobial susceptibility of two indicator bacteria, E. coli and Enterococcus spp, isolated from otter spraints and interpreted the results according to clinical and epidemiological cut-offs (ECOFFs).