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.

Endoparasites are the potential source of substantial health complications in animals; exclusively the endoparasites of zoonotic importance are of great concern to researchers and health authorities for diverse perspectives. A coprological study was conducted to inspect the endoparasitic infestation in antelopes kept at three captive localities, i.e., Safari Park, Jallo Wildlife Park, and Lahore Zoo, situated in Lahore, Pakistan. There were 109 selected species of antelopes including nilgai (Boselaphus tragocamelus), blackbuck (Antilope cervicapra), urial (Ovis orientalis), and chinkara (Gazella bennettii). The fresh fecal samples of each experimental animal from individual and mixed animal enclosures were collected and tested by an appropriate parasitological method. The fecal samples were examined by applying the modified McMaster technique through the Whitlock chamber method and observed by a compound microscope for identification and fecal egg count. The prevalence (%) of egg per gram (EPG) and diversity of endoparasitic eggs identified from fecal samples of experimental animals from three captive localities were recorded. The analysis revealed species from phylogenetic groups of nematodes cestodes and trematodes along with coccidian occurrence. The prevalence of endoparasites was highest in nilgai (B. tragocamelus) with combined average prevalence (23.88 ± 3.13) from three captive localities followed by chinkara (G. bennettii) combined average prevalence (21.68 ± 2.64), urial (O. orientalis) combined average prevalence (21.41 ± 4.69), and blackbuck (A. cervicapra) combined average prevalence (16.88 ± 2.66). To prevent such infestations which prevail more intensely regarding changing climate and increasing pollution levels, there should be regular monitoring and appropriate prophylaxis combined with epizootiological investigation for future studies and implication of advance technology, for captive animals so that best possible adaptations can be made to reduce the spread of infective diseases that are of zoonotic importance also.