Extended-spectrum β-lactamases (ESBL)-producing Enterobacterales have been classified as critical priority pathogens by the World Health Organization (WHO). ESBL are universally distributed and, in 2006, were firstly reported on a wild animal. Understanding the relative contributions of wild animals to ESBL circulation in the environment is urgently needed. In this work, we have conducted a nationwide study in Portugal to investigate the occurrence of bacteria carrying clinically significant antimicrobial resistance genes (ARG), using widely distributed wild ungulates as model species. A total of 151 antimicrobial resistant-Enterobacterales isolates were detected from 181 wild ungulates: 50% (44/88) of isolates from wild boar (Sus scrofa), 40.3% (25/62) from red deer (Cervus elaphus), 41.4% (12/29) from fallow deer (Dama dama) and 100% (2/2) from mouflon (Ovis aries subsp. musimon). Selected isolates showed a diversified resistance profile, with particularly high values corresponding to ampicillin (71.5%) and tetracycline (63.6%). Enterobacterales strains carried blaTEM, tetA, tetB, sul2, sul1 or dfrA1 ARG genes. They also carried blaCTX₋M-type genes, which are prevalent in human infections, namely CTX-M-14, CTX-M-15 and CTX-M-98. Strikingly, this is the first report of CTX-M-98 in wildlife. Almost 40% (n = 59) of Enterobacterales were multi-drug resistant. The diversity of plasmids carried by ESBL isolates was remarkable, including IncF, K and P. This study highlights the potential role of wild ungulates as environmental reservoirs of CTX-M ESBL-producing E. coli and in the spill-over of AMR bacteria and their determinants. Our findings suggest that wild ungulates are useful as strategic sentinel species of AMR in terrestrial environments, especially in response to potential sources of anthropogenic pollution, providing early warning of potential risks to human, animal and environmental health.

Endotoxemia is mainly caused by translocation of bacterial lipopolysaccharides (LPS) into the bloodstream. This in turn enhances systemic inflammation and inappropriate production of reactive oxygen species, leading to oxidative injury of vital internal organs and other dangerous effects that can be life-threatening. Here, we evaluated/compared the modulatory effects of consuming two different doses (2% and 4% of the diet) of brown seaweeds (Sargassum latifolium) for 40 consecutive days on thermo-respiratory response, inflammation, and oxidative stress in Barki male sheep (Ovis aries) challenged twice with bacterial LPS (1.25 μg/kg body weight, injected intravenously on days 28 and 35 of the experimental period). The results showed that the diet containing Sargassum latifolium (especially at 4%) modulated significantly (P < 0.05–0.001) the increase in the thermo-respiratory response (skin and rectal temperatures, and respiration rate) and the obtained systemic inflammation (blood leukocytosis, the elevation in the erythrocyte sedimentation rate, and the increase in serum proinflammatory cytokines and heat shock protein-70 concentrations) in the LPS-challenged sheep. In addition, it improved significantly (P < 0.001, especially at 4%) the total antioxidant capacity of the blood of LPS-challenged sheep by increasing the catalase and superoxide dismutase activities. Moreover, it decreased the blood markers of tissue damage (malondialdehyde concentration and the activities of alanine aminotransferase and lactate dehydrogenase) in the LPS-challenged sheep. In conclusion, the diet containing 4% Sargassum latifolium may have potential impact in protecting the ruminant livestock from the serious effects of endotoxemia through improving the animals’ antioxidant defense system and regulating their inflammatory and thermo-respiratory responses.