Spatial and temporal variation in the prevalence of illegal lead shot in reared and wild mallards harvested in England.

The use of lead shotgun ammunition for shooting wildfowl has been restricted in England since 1999, but surveys finding lead shot in harvested birds show compliance with regulations has been low. Following the announcement in 2020 of a voluntary transition from lead to non-lead shot by UK shooting organizations, we investigated spatiotemporal variation in the composition of ammunition used for shooting mallards Anas platyrhynchos. We collected 176 harvested mallards during the 2021/22 shooting season and analyzed recent shot extracted from carcasses to determine shot composition. Using a separate collection of ducks of known provenance, we used stable isotope analysis as a means of differentiating captive-reared from wild mallards. This allowed us to understand how compliance might vary between driven game shooters, characterized by shooting birds that are flushed over a stationary line of shooters, and who primarily harvest captive-reared and released ducks, and wild duck shooters. Of 133 mallards containing recent shot, 92 (69%) had been illegally shot with lead. Analysis of this and five comparable surveys between 2001 and 2019 indicates regional and temporal variation in lead shot presence in England. In the North West and West Midlands, the likelihood of mallards containing lead shot decreased significantly over time, but no other regions showed significant changes. The use of non-lead shot types varied over time, with increases in steel shot use approximately matched by declines in bismuth shot. Mallards likely to be reared were more likely to have been shot with lead (75%) than those likely to be wild (48%). This suggests the use of lead shot is more frequent among driven game shooters than wild duck shooters. In England in 2021/22, most mallards continued to be shot with lead, suggesting that neither legislation nor voluntary approaches have been effective in substantially reducing illegal use of lead shot.

Sarcocystis rileyi in UK free-living wildfowl (Anatidae): surveillance, histopathology and first molecular characterisation.

BACKGROUND: Reports from UK hunters of 'rice grains' in muscles of shot wildfowl (Anatidae) coincided temporally with the finding of sarcocystosis in a number of ducks found as part of the Wildfowl & Wetlands Trust long-term general surveillance of found dead waterbirds. Sarcocystis rileyi has also been relatively recently confirmed in wildfowl in north-eastern Europe. METHODS: This study uses four approaches to investigate UK wildfowl sarcocystosis: first, through a hunter questionnaire that captured historical case data; secondly, through an online reporting system; thirdly, DNA sequencing to characterise UK cases; and fourthly, histological myopathy assessment of infected pectoral muscle. RESULTS: Our questionnaire results suggest Sarcocystis infection is widely distributed throughout the UK and observed in 10 Anatidae species, reported cases increased since the 2010/2011 shooting season, with the online reporting system reflecting this increase. DNA sequencing (18S rRNA gene and internal transcribed spacer-1 region) of UK isolates confirmed S rileyi in the five dabbling duck host species tested and the associated histopathological myopathy is described. CONCLUSION: This work highlights an emerging issue to European wildfowl species and provides much opportunity for further research, including the impacts of S rileyi and the described myopathy on host health, fitness and survival.

The Evolution of Innate Immune Genes: Purifying and Balancing Selection on ß-Defensins in Waterfowl.

In disease dynamics, high immune gene diversity can confer a selective advantage to hosts in the face of a rapidly evolving and diverse pathogen fauna. This is supported empirically for genes involved in pathogen recognition and signalling. In contrast, effector genes involved in pathogen clearance may be more constrained. ß-Defensins are innate immune effector genes; their main mode of action is via disruption of microbial membranes. Here, five ß-defensin genes were characterized in mallards (Anas platyrhynchos) and other waterfowl; key reservoir species for many zoonotic diseases. All five genes showed remarkably low diversity at the individual-, population-, and species-level. Furthermore, there was widespread sharing of identical alleles across species divides. Thus, specific ß-defensin alleles were maintained not only spatially but also over long temporal scales, with many amino acid residues being fixed across all species investigated. Purifying selection to maintain individual, highly efficacious alleles was the primary evolutionary driver of these genes in waterfowl. However, we also found evidence for balancing selection acting on the most recently duplicated ß-defensin gene (AvBD3b). For this gene, we found that amino acid replacements were more likely to be radical changes, suggesting that duplication of ß-defensin genes allows exploration of wider functional space. Structural conservation to maintain function appears to be crucial for avian ß-defensin effector molecules, resulting in low tolerance for new allelic variants. This contrasts with other types of innate immune genes, such as receptor and signalling molecules, where balancing selection to maintain allelic diversity has been shown to be a strong evolutionary force.

Potential hazard to human health from exposure to fragments of lead bullets and shot in the tissues of game animals.

BACKGROUND: Lead is highly toxic to animals. Humans eating game killed using lead ammunition generally avoid swallowing shot or bullets and dietary lead exposure from this source has been considered low. Recent evidence illustrates that lead bullets fragment on impact, leaving small lead particles widely distributed in game tissues. Our paper asks whether lead gunshot pellets also fragment upon impact, and whether lead derived from spent gunshot and bullets in the tissues of game animals could pose a threat to human health. METHODOLOGY/PRINCIPAL FINDINGS: Wild-shot gamebirds (6 species) obtained in the UK were X-rayed to determine the number of shot and shot fragments present, and cooked using typical methods. Shot were then removed to simulate realistic practice before consumption, and lead concentrations determined. Data from the Veterinary Medicines Directorate Statutory Surveillance Programme documenting lead levels in raw tissues of wild gamebirds and deer, without shot being removed, are also presented. Gamebirds containing > or =5 shot had high tissue lead concentrations, but some with fewer or no shot also had high lead concentrations, confirming X-ray results indicating that small lead fragments remain in the flesh of birds even when the shot exits the body. A high proportion of samples from both surveys had lead concentrations exceeding the European Union Maximum Level of 100 ppb w.w. (0.1 mg kg(-1) w.w.) for meat from bovine animals, sheep, pigs and poultry (no level is set for game meat), some by several orders of magnitude. High, but feasible, levels of consumption of some species could result in the current FAO/WHO Provisional Weekly Tolerable Intake of lead being exceeded. CONCLUSIONS/SIGNIFICANCE: The potential health hazard from lead ingested in the meat of game animals may be larger than previous risk assessments indicated, especially for vulnerable groups, such as children, and those consuming large amounts of game.

PCR analysis of the presence and location of Mycobacterium avium in a constructed reed bed, with implications for avian tuberculosis control.

The potential of reed beds to act as biofilters of pathogenic and environmental mycobacteria was investigated through examination of the fate of mycobacteria in a constructed reed bed filtering effluent from a large captive wildfowl collection. Particular emphasis was placed on the presence and location of Mycobacterium avium--the causal agent of avian tuberculosis (ATB)--in an effort to clarify the potential role of reed beds in the control of this disease. Water, sediment, and stems and roots of common reed (Phragmites australis) and greater reedmace (Typha latifolia) were taken from 15 locations within the reed bed plus sites upstream and downstream. Samples were analysed for mycobacteria using PCR and specifically for M. avium using nested PCR. Environmental mycobacteria were found throughout the entire reed bed but M. avium was not found downstream of the first vegetation growth. The reed bed was found to effectively remove M. avium from the water through a combination of sedimentation and adsorption onto vegetation stems. The results of this study show that constructed reed beds composed of a settlement lagoon and one or more vegetation beds can act as valuable and ecologically friendly tools in the environmental control of ATB.