Consumption of meat from animals hunted with Pb ammunition can cause toxic accumulation with consequent health risks, even if relatively small amounts are consumed in each exposure. In El Palmar National Park, Argentina, invasive alien mammals, wild boar (Sus scrofa) and axis deer (Axis axis), are culled with Pb ammunition and their meat is consumed. In this study, we evaluated blood Pb concentrations in 58 consumers of culled game and examined Pb exposure risk according to their demographics, duty, and consumption habits. Likewise, the likelihood of exposure was evaluated by quantifying Pb concentrations in meat samples of seven culled axis deer. Twenty-seven participants (46%) had detectable blood Pb levels (limit of detection = 3.3 μg/dL), with an average 4.75 ± 1.35 μg/dL (geometric mean ± geometric S.D.); the average for all participants was 3.25 ± 1.51 μg/dL. Blood Pb concentrations were significantly higher in hunters, in participants who reported consuming game meat more than 5 times per week, and in participants who reported frequently consuming cured game meat (compared to cooked or pickled). Pb concentration varied significantly along the trajectory of the bullet in deer muscle, being highest at mid-point but with detectable Pb levels even in distant tissue samples (control), suggesting potential for dietary intake by consumers. These findings provide evidence of Pb exposure risk in consumers and emphasize the relevance of replacing Pb ammunition with non-toxic alternatives. This change would reduce dietary exposure in frequent consumers and allow the use of game meat as safe food for people whilst eliminating collateral risks to wild animals and the environment.
The hepatitis E virus (HEV) is an emergent zoonotic virus causing viral hepatitis worldwide. Clinically, hepatitis E is not easily distinguished from other types of acute viral hepatitis. There is a need for HEV diagnostic assays to detect and prevent interspecies transmission among susceptible populations. Nanobodies (Nbs) are expressed recombinantly in different systems, produced with high yields, and have superior physicochemical properties compared with conventional antibodies (Ab). Several Nbs against ORF2, the capsid protein and main antigen, were selected and produced in E. coli. Nb39 and Nb74 specifically recognized HEV ORF2 (genotypes 3 and 4). A competitive ELISA (cELISA) was developed and validated using a reference panel of human (n = 86) and swine sera (n = 116) tested in comparison with a commercial kit. The optimal cutoff values determined by ROC analysis were 69.16% (human) and 58.76% (swine); the sensitivity and specificity were high: 97.4% (IC95%; 86.5–99.5%) and 95.8% (IC95%; 86.0-98.8%) for human vs. 100% (IC95%: 93.5–100%) and 98.3% (IC95%: 91.0-99.7%) for swine. Further, the cELISA detected total anti-HEV antibodies in wild boar, deer, and mice. To our knowledge, this is the first report of production of Nbs against HEV-3 ORF2 for diagnostic purposes.
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