Winged resistance: Storks and gulls increase carriage of antibiotic resistance by shifting from paddy fields to landfills.

Waterbirds are vectors for the dissemination of antimicrobial resistance across environments, with some species increasingly reliant on highly anthropized habitats for feeding. However, data on the impact of their feeding habits on the carriage of antibiotic resistance genes (ARGs) are still scarce. To fill this gap, we examined the microbiota (16S rRNA amplicon gene sequencing) and the prevalence of ARG (high-throughput qPCR of 47 genes) in faeces from white storks (Ciconia ciconia) and lesser black-backed gulls (Larus fuscus) feeding in highly (landfill) and less (paddy fields) polluted habitats. Faecal bacterial richness and diversity were higher in gulls feeding upon landfills and showed a greater abundance of potential pathogens, such as Staphylococcus. In contrast, faecal bacterial communities from storks were similar regardless of habitat preferences, maybe due to a less intense habitat use compared to gulls. In addition, birds feeding in the landfill carried a higher burden of ARGs compared to the surrounding soil and surface waters. Network analysis revealed strong correlations between ARGs and potential pathogens, particularly between tetM (resistance to tetracyclines), blaCMY (beta-lactam resistance), sul1 (sulfonamide resistance) and members of the genera Streptococcus, Peptostreptococcus, and Peptoclostridium. Our work demonstrates how transitioning from paddy fields to landfills fosters the carriage of ARGs and potential pathogens in the bird gut, shedding light on the ecological role of these avian vectors in antimicrobial resistance dissemination.

Biovectoring of plastic by white storks from a landfill to a complex of salt ponds and marshes.

Research into plastic pollution has extensively focused on abiotic vectors, overlooking transport by animals. Opportunistic birds, such as white storks (Ciconia ciconia) often forage on landfills, where plastic abounds. We assess plastic loading by ingestion and regurgitation of landfill plastic in Cadiz Bay, a major stopover area for migratory white storks in south-west Spain. On average, we counted 599 storks per day moving between a landfill and a complex of salt ponds and marshes, where they regurgitated pellets that each contained a mean of 0.47 g of plastic debris, dominated by polyethylene. Modelling reliant on GPS tracking estimated that 99 kg and >2 million particles of plastic were biovectored into the wetland during 2022, with seasonal peaks that followed migration patterns. GPS data enabled the correction of field censuses and the identification of plastic deposition hotspots. This study highlights the important role that biovectoring plays in plastic transport into coastal wetlands.

Seed Size, Not Dispersal Syndrome, Determines Potential for Spread of Ricefield Weeds by Gulls.

Recent field data suggest that migratory gulls disperse many rice field weeds by gut passage (endozoochory), most of which are dry fruited and widely assumed to have no long-distance dispersal mechanisms, except via human activity. We investigated this mechanism with a feeding experiment, in which seeds of five common rice field weeds (in order of increasing seed size: Juncus bufonius, Cyperus difformis, Polypogon monspeliensis, Amaranthus retroflexus, and the fleshy-fruited Solanum nigrum) were fed to seven individuals of lesser black-backed gulls Larus fuscus held in captivity. We quantified seed survival after collecting faeces at intervals for 33 h after ingestion, then extracting intact seeds and running germination tests, which were also conducted for control seeds. All five species showed high seed survival after gut passage, of >70%. Gut retention times averaged 2-4 h, but maxima exceeded 23 h for all species. Germinability after gut passage was 16-54%, and gut passage accelerated germination in J. bufonius and S. nigrum, but slowed it down in the other species. All species had lower germinability after gut passage compared to control seeds (likely due to stratification prior to the experiment), but the loss of germinability was higher in smaller seeds. There was no evidence that the different dispersal syndromes assigned to the five species (endozoochory, epizoochory or barochory) had any influence on our results. In contrast, mean gut retention time was strongly and positively related to seed size, likely because small seeds pass more quickly from the gizzard into the intestines. Non-classical endozoochory of dry-fruited seeds by waterbirds is a major but overlooked mechanism for potential long-distance dispersal, and more research into this process is likely essential for effective weed management.

Trace element bioaccumulation in hypersaline ecosystems and implications of a global invasion.

Hypersaline ecosystems are under increasing threat due to anthropogenic pressures such as environmental pollution and biological invasions. Here we address the ecotoxicological implications of the Artemia franciscana (Crustacea) invasion in saltpans of southern Spain. This North American species is causing the extinction of native Artemia populations in many parts of the globe. The bioaccumulation of trace elements (As, Cd, Cu, Co, Cr, Mn, Ni, Pb and Zn) in native populations (A. parthenogenetica) from Cabo de Gata and Odiel saltpans and invasive Artemia from Cádiz saltpan was studied at different salinities. Furthermore, in Odiel, the most polluted study site, we also analysed the bioaccumulation of trace elements by Chironomus salinarius larvae (Diptera) and Ochthebius notabilis adults (Coleoptera). High levels of trace elements were detected in the studied saltpans, many of them exceeding the recommended threshold guidelines for aquatic life. Bioaccumulation of trace elements by Artemia was lowest at the highest salinity. The invasive A. franciscana showed higher potential to bioaccumulate trace elements than its native counterpart (in particular for As, Cd, Ni and Cr). In Odiel, O. notabilis stood out as showing the highest potential to bioaccumulate As and Cu. Results showed that the shift from a native to an alien Artemia species with a higher bioaccumulation capacity may increase the transfer of trace elements in hypersaline food webs, especially for waterbirds that depend on Artemia as food. Thus, our study identifies an indirect impact of the Artemia franciscana invasion that had not previously been recognised.

Faecal microbiota and antibiotic resistance genes in migratory waterbirds with contrasting habitat use.

Migratory birds may have a vital role in the spread of antimicrobial resistance across habitats and regions, but empirical data remain scarce. We investigated differences in the gut microbiome composition and the abundance of antibiotic resistance genes (ARGs) in faeces from four migratory waterbirds wintering in South-West Spain that differ in their habitat use. The white stork Ciconia ciconia and lesser black-backed gull Larus fuscus are omnivorous and opportunistic birds that use highly anthropogenic habitats such as landfills and urban areas. The greylag goose Anser anser and common crane Grus grus are herbivores and use more natural habitats. Fresh faeces from 15 individuals of each species were analysed to assess the composition of bacterial communities using 16S rRNA amplicon-targeted sequencing, and to quantify the abundance of the Class I integron integrase gene (intI1) as well as genes encoding resistance to sulfonamides (sul1), beta-lactams (blaTEM, blaKPC and blaNDM), tetracyclines (tetW), fluoroquinolones (qnrS), and colistin (mcr-1) using qPCR. Bacterial communities in gull faeces were the richest and most diverse. Beta diversity analysis showed segregation in faecal communities between bird species, but those from storks and gulls were the most similar, these being the species that regularly feed in landfills. Potential bacterial pathogens identified in faeces differed significantly between bird species, with higher relative abundance in gulls. Faeces from birds that feed in landfills (stork and gull) contained a significantly higher abundance of ARGs (sul1, blaTEM, and tetW). Genes conferring resistance to last resort antibiotics such as carbapenems (blaKPC) and colistin (mcr-1) were only observed in faeces from gulls. These results show that these bird species are reservoirs of antimicrobial resistant bacteria and suggest that waterbirds may disseminate antibiotic resistance across environments (e.g., from landfills to ricefields or water supplies), and thus constitute a risk for their further spread to wildlife and humans.

Life history and physiological responses of native and invasive brine shrimps exposed to zinc.

Although a substantial amount of research exists on pollution and biological invasions, there is a paucity of understanding of how both factors interact. Most studies show that pollution favours the establishment of invasive species, but pollution may also promote local adaptation of native species and prevent the establishment of new incomers. However, evidence for this is extremely limited because most studies focus on successful invasions and very few on cases where an invasion has been resisted. Here we provide evidence of local adaptation of native species to pollution combining life history and physiological data. We focused on the invasion of the North American brine shrimp Artemia franciscana, which is causing a dramatic biodiversity loss in hypersaline ecosystems worldwide, and one of the last native Artemia populations in SW Europe (A. parthenogenetica from the historically polluted Odiel estuary, SW Spain). Life table response experiments were carried out in the laboratory to compare the demographic responses of A. parthenogenetica and a nearby A. franciscana population to long-term Zn exposure (0.2 mg L-1). We also evaluated oxidative stress by measuring antioxidant defences (catalase, glutathione reductase and superoxide dismutase) and lipid peroxidation (thiobarbituric acid reactive substances). A high concentration of Zn induced strong mortality in A. franciscana, which also showed high levels of lipid peroxidation, suggesting relatively poor physiological resistance to pollution compared with A. parthenogenetica. The age at maturity was shorter in A. parthenogenetica, which may be an adaptation to the naturally high mortality rate observed in the Odiel population. Exposure to Zn accelerated age at first reproduction in A. franciscana but not in A. parthenogenetica. In contrast, Zn had a stimulatory effect on offspring production in A. parthenogenetica,which also showed higher reproductive parameters (number of broods, total offspring and offspring per brood) than A. franciscana. Overall, the results of this study strongly suggest that native Artemia from Odiel estuary is locally adapted (at both, reproductive and physiological levels) to Zn contamination and that A. franciscana is highly sensitive. This is a good example of how pollution may play a role in the persistence of the last native Artemia populations in the Mediterranean.

Larval helminths in the invasive American brine shrimp Artemia franciscana throughout its annual cycle.

One of the best examples of rapid displacement of native species by an invader is the eradication of native Artemia salina and A. parthenogenetica in the Mediterranean by the introduced American A. franciscana. Previous studies based on sampling from limited time periods suggest that the success of the American species as a competitor may be due partly to different parasite burden, since native Artemia spp. have high cestode infection rates regulating their density. The aim of this study is to test the hypothesis that the helminth infection in A. franciscana in its invasive range is low throughout its annual life cycle. Samples of A. franciscana were collected every second month from La Tapa saltern (Andalusia) during one year. Five helminth species were recorded: cestodes Flamingolepis liguloides, F. flamingo, Gynandrotaenia stammeri (all flamingo parasites), Eurycestus avoceti (a shorebird parasite) and larval spirurids of the Acuariinae (the first record of nematodes in Artemia). The overall infection rate was low, with total prevalence 5.9% and prevalence of individual parasite species between 0.2 and 3.2%. The mean abundance of helminths was 0.005-0.155 (av. 0.068), 5-13 times lower than in native congeners. Waterbird counts indicate that the low infection rates cannot be explained by lack of definitive hosts. The results are consistent with the hypothesis that helminths have no regulating effect on the invasive brine shrimp in the Mediterranean. The replacement of the native populations by the invader can be partially explained by a competition mediated by parasites/predators through a differential impact on host fitness.

Interannual variations in feeding frequencies and food quality of greater flamingo chicks (Phoenicopterus roseus): evidence from plasma chemistry and effects on body condition.

Greater flamingos in southern Spain foraged in areas distant from a breeding site, spending 4-6 days in foraging areas between successive visits to the colony to feed their chicks. During four years, we took blood samples from chicks to ascertain whether there were interannual variations in several blood parameters, indicative of food quality and feeding frequencies. When the chicks were captured, 20-31% of them had their crops empty, indicating that not all chicks were fed daily. Additional evidence of variations in feeding frequencies was obtained from a principal component analysis (PCA) on plasma chemistry values, which also indicated that there were annual variations in the quality of food received by chicks. The association of cholesterol and glucose with some PC axes indicated that some chicks were experiencing fasting periods. Of all plasma metabolites considered, cholesterol was the best one to predict body condition. Greater flamingo chicks experiencing longer fasting intervals, as suggested by higher plasma levels of cholesterol, were in lower body condition.