Experimental evidence of dispersal of invasive cyprinid eggs inside migratory waterfowl.

Fish have somehow colonized isolated water bodies all over the world without human assistance. It has long been speculated that these colonization events are assisted by waterbirds, transporting fish eggs attached to their feet and feathers, yet empirical support for this is lacking. Recently, it was suggested that endozoochory (i.e., internal transport within the gut) might play a more important role, but only highly resistant diapause eggs of killifish have been found to survive passage through waterbird guts. Here, we performed a controlled feeding experiment, where developing eggs of two cosmopolitan, invasive cyprinids (common carp, Prussian carp) were fed to captive mallards. Live embryos of both species were retrieved from fresh feces and survived beyond hatching. Our study identifies an overlooked dispersal mechanism in fish, providing evidence for bird-mediated dispersal ability of soft-membraned eggs undergoing active development. Only 0.2% of ingested eggs survived gut passage, yet, given the abundance, diet, and movements of ducks in nature, our results have major implications for biodiversity conservation and invasion dynamics in freshwater ecosystems.

Genomic data support multiple introductions and explosive demographic expansions in a highly invasive aquatic insect.

The study of the genetic makeup and demographic fate of alien species is essential to understand their capacity to recover from founder effects, adapt to new environmental conditions and, ultimately, become invasive and potentially damaging. Here, we employ genomic data to gain insights into key demographic processes that might help to explain the extraordinarily successful invasion of the Western Mediterranean region by the North American boatman Trichocorixa verticalis (Hemiptera: Corixidae). Our analyses revealed the genetic distinctiveness of populations from the main areas comprising the invasive range and coalescent-based simulations supported that they originated from independent introductions events probably involving different source populations. Testing of alternative demographic models indicated that all populations experienced a strong bottleneck followed by a recent and instantaneous demographic expansion that restored a large portion (>30%) of their ancestral effective population sizes shortly after introductions took place (<60 years ago). Considerable genetic admixture of some populations suggest that hypothetical barriers to dispersal (i.e., land and sea water) are permeable to gene flow and/or that they originated from introductions involving multiple lineages. This study demonstrates the repeated arrival of propagules with different origins and short time lags between arrival and establishment, emphasizing the extraordinary capacity of the species to recover from founder effects and genetically admix in invaded areas. This can explain the demonstrated capacity of this aquatic insect to spread and outcompete native species once it colonizes new suitable regions. Future genomic analyses of native range populations could help to infer the genetic makeup of introduced populations and track invasion routes.

Adaptive introgression of the beta-globin cluster in two Andean waterfowl.

Introgression of beneficial alleles has emerged as an important avenue for genetic adaptation in both plant and animal populations. In vertebrates, adaptation to hypoxic high-altitude environments involves the coordination of multiple molecular and cellular mechanisms, including selection on the hypoxia-inducible factor (HIF) pathway and the blood-O2 transport protein hemoglobin (Hb). In two Andean duck species, a striking DNA sequence similarity reflecting identity by descent is present across the ~20 kb ß-globin cluster including both embryonic (HBE) and adult (HBB) paralogs, though it was yet untested whether this is due to independent parallel evolution or adaptive introgression. In this study, we find that identical amino acid substitutions in the ß-globin cluster that increase Hb-O2 affinity have likely resulted from historical interbreeding between high-altitude populations of two different distantly-related species. We examined the direction of introgression and discovered that the species with a deeper mtDNA divergence that colonized high altitude earlier in history (Anas flavirostris) transferred adaptive genetic variation to the species with a shallower divergence (A. georgica) that likely colonized high altitude more recently possibly following a range shift into a novel environment. As a consequence, the species that received these ß-globin variants through hybridization might have adapted to hypoxic conditions in the high-altitude environment more quickly through acquiring beneficial alleles from the standing, hybrid-origin variation, leading to faster evolution.

The future for Mediterranean wetlands: 50 key issues and 50 important conservation research questions.

Wetlands are critically important for biodiversity and human wellbeing, but face a range of challenges. This is especially true in the Mediterranean region, where wetlands support endemic and threatened species and remain integral to human societies, but have been severely degraded in recent decades. Here, in order to raise awareness of future challenges and opportunities for Mediterranean wetlands, and to inform proactive research and management, we identified (a) 50 key issues that might affect Mediterranean wetlands between 2020 and 2050, and (b) 50 important research questions that, if answered, would have the greatest impact on the conservation of Mediterranean wetlands between 2020 and 2050. We gathered ideas through an online survey and review of recent literature. A diverse assessment panel prioritised ideas through an iterative, anonymised, Delphi-like process of scoring, voting and discussion. The prioritised issues included some that are already well known but likely to have a large impact on Mediterranean wetlands in the next 30 years (e.g. the accumulation of dams and reservoirs, plastic pollution and weak governance), and some that are currently overlooked in the context of Mediterranean wetlands (e.g. increasing desalination capacity and development of antimicrobial resistance). Questions largely focused on how best to carry out conservation interventions, or understanding the impacts of threats to inform conservation decision-making. This analysis will support research, policy and practice related to environmental conservation and sustainable development in the Mediterranean, and provides a model for similar analyses elsewhere in the world. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10113-020-01743-1.

When Parasites Are Good for Health: Cestode Parasitism Increases Resistance to Arsenic in Brine Shrimps.

Parasites and pollutants can both affect any living organism, and their interactions can be very important. To date, repeated studies have found that parasites and heavy metals or metalloids both have important negative effects on the health of animals, often in a synergistic manner. Here, we show for the first time that parasites can increase host resistance to metalloid arsenic, focusing on a clonal population of brine shrimp from the contaminated Odiel and Tinto estuary in SW Spain. We studied the effect of cestodes on the response of Artemia to arsenic (acute toxicity tests, 24h LC50) and found that infection consistently reduced mortality across a range of arsenic concentrations. An increase from 25°C to 29°C, simulating the change in mean temperature expected under climate change, increased arsenic toxicity, but the benefits of infection persisted. Infected individuals showed higher levels of catalase and glutathione reductase activity, antioxidant enzymes with a very important role in the protection against oxidative stress. Levels of TBARS were unaffected by parasites, suggesting that infection is not associated with oxidative damage. Moreover, infected Artemia had a higher number of carotenoid-rich lipid droplets which may also protect the host through the "survival of the fattest" principle and the antioxidant potential of carotenoids. This study illustrates the need to consider the multi-stress context (contaminants and temperature increase) in which host-parasite interactions occur.

Convergent Evolution of Hemoglobin Function in High-Altitude Andean Waterfowl Involves Limited Parallelism at the Molecular Sequence Level.

A fundamental question in evolutionary genetics concerns the extent to which adaptive phenotypic convergence is attributable to convergent or parallel changes at the molecular sequence level. Here we report a comparative analysis of hemoglobin (Hb) function in eight phylogenetically replicated pairs of high- and low-altitude waterfowl taxa to test for convergence in the oxygenation properties of Hb, and to assess the extent to which convergence in biochemical phenotype is attributable to repeated amino acid replacements. Functional experiments on native Hb variants and protein engineering experiments based on site-directed mutagenesis revealed the phenotypic effects of specific amino acid replacements that were responsible for convergent increases in Hb-O2 affinity in multiple high-altitude taxa. In six of the eight taxon pairs, high-altitude taxa evolved derived increases in Hb-O2 affinity that were caused by a combination of unique replacements, parallel replacements (involving identical-by-state variants with independent mutational origins in different lineages), and collateral replacements (involving shared, identical-by-descent variants derived via introgressive hybridization). In genome scans of nucleotide differentiation involving high- and low-altitude populations of three separate species, function-altering amino acid polymorphisms in the globin genes emerged as highly significant outliers, providing independent evidence for adaptive divergence in Hb function. The experimental results demonstrate that convergent changes in protein function can occur through multiple historical paths, and can involve multiple possible mutations. Most cases of convergence in Hb function did not involve parallel substitutions and most parallel substitutions did not affect Hb-O2 affinity, indicating that the repeatability of phenotypic evolution does not require parallelism at the molecular level.

Great cormorants reveal overlooked secondary dispersal of plants and invertebrates by piscivorous waterbirds.

In wetland ecosystems, birds and fish are important dispersal vectors for plants and invertebrates, but the consequences of their interactions as vectors are unknown. Darwin suggested that piscivorous birds carry out secondary dispersal of seeds and invertebrates via predation on fish. We tested this hypothesis in the great cormorant (Phalacrocorax carbo L.). Cormorants regurgitate pellets daily, which we collected at seven European locations and examined for intact propagules. One-third of pellets contained at least one intact plant seed, with seeds from 16 families covering a broad range of freshwater, marine and terrestrial habitats. Of 21 plant species, only two have an endozoochory dispersal syndrome, compared with five for water and eight for unassisted dispersal syndromes. One-fifth of the pellets contained at least one intact propagule of aquatic invertebrates from seven taxa. Secondary dispersal by piscivorous birds may be vital to maintain connectivity in meta-populations and between river catchments, and in the movement of plants and invertebrates in response to climate change. Secondary dispersal pathways associated with complex food webs must be studied in detail if we are to understand species movements in a changing world.

Immigration enhances fast growth of a newly established source population.

Abstract. Immigration and local recruitment play a central role in determining the growth rate of breeding populations. Unraveling these processes in newly established pop- ulations is of great importance to increase our understanding of how species change their distributions in response to global change. We studied the largest colony of glossy ibis (Plegadis falcinellus) in Western Europe (established in 1996 in Doñana, SW-Spain) by using capture-recapture methods, count estimates, and projection matrix modeling to: (1) test the effect of resource availability and competition on local recruitment dynamics, (2) investigate the contribution of local recruitment vs. immigration on population growth, and (3) assess the role of this population in source/sink dynamics. We found different dynamics before and after the establishment of satellite colonies in Doñana in 2004. Between 1996 and 2003, the population increased rapidly, fueled by immigrants (≈ 58 breeding females/ yr). Between 2003 and 2007, however, both colony size increase and immigration were negligible. Immigration played a major role in colony growth, but simultaneously this colony was a source population driving expansion of the species range as suggested by (1) absolute and relative estimates of the observed growth rate relative to that predicted by self-recruitment, and (2) numerous observations of Doñana-born individuals breeding elsewhere. Local recruitment, which was particularly high for first-year individuals (probability > 0.8 for the early study years), was not directly related to resource availability or previous-year breeding success. Local recruitment decreased rapidly at a threshold population size, however, when other satellite colonies became established at Dofiana. Our study suggests that even when recruitment at an early age and high productivity are observed, immigration can still play a pivotal role in promoting the fast growth of new populations at the edge of a species range, at least until density-dependent effects arise. This process can be so fast that within a few years a new population can itself become a source population, further driving range expansion of the species.

Waterfowl endozoochory: An overlooked long-distance dispersal mode for Cuscuta (dodder).

PREMISE OF THE STUDY: Dispersal of parasitic Cuscuta species (dodders) worldwide has been assumed to be largely anthropomorphic because their seeds do not match any previously known dispersal syndrome and no natural dispersal vectors have been reliably documented. However, the genus has a subcosmopolitan distribution and recent phylogeographic results have indicated that at least18 historical cases of long-distance dispersal (LDD) have occurred during its evolution. The objective of this study is to report the first LDD biological vector for Cuscuta seeds. METHODS: Twelve northern pintails (Anas acuta) were collected from Suisun Marsh, California and the contents of their lowest part of the large intestine (rectum) were extracted and analyzed. Seed identification was done both morphologically and using a molecular approach. Extracted seeds were tested for germination and compared to seeds not subjected to gut passage to determine the extent of structural changes caused to the seed coat by passing through the digestive tract. KEY RESULTS: Four hundred and twenty dodder seeds were found in the rectum of four northern pintails. From these, 411 seeds were identified as Cuscuta campestris and nine as most likely C. pacifica. The germination rate of C. campestris seeds after gut passage was 55%. Structural changes caused by the gut passage in both species were similar to those caused by an acid scarification. CONCLUSIONS: Endozoochory by waterbirds may explain the historical LDD cases in the evolution of Cuscuta. This also suggests that current border quarantine measures may be insufficient to stopping spreading of dodder pests along migratory flyways.

Risk assessment of lead poisoning and pesticide exposure in the declining population of red-breasted goose (Branta ruficollis) wintering in Eastern Europe.

The red-breasted goose Branta ruficollis is a globally threatened species (IUCN Vulnerable) and the only European goose species currently in decline. Working on the wintering grounds on the Black Sea Coast, we address two potential causes of decline of this species for the first time: lead poisoning, and contamination from pesticides. We quantified the densities of spent Pb shot in three wetlands used by the geese in north-east Bulgaria, and analysed the Pb concentration in the faeces of red-breasted geese and the more abundant greater white-fronted geese Anser albifrons, using Al concentration as an indicator of soil ingestion. Pb shot densities in sediments were low, and we found no evidence for Pb shot ingestion in red-breasted geese. On the other hand, we found that the geese were feeding on wheat whose seeds were treated with four fungicides: thiram, tebuconazole, difenoconazole and fludioxonil, and the two first were even detected in geese faecal samples. Using data on the daily food intake, we estimated the exposure levels of the geese to these fungicides, both by measuring the concentrations remaining on seeds and by estimating the amount used to coat the seeds at the time of sowing. We found that the exposure rates estimated during the sowing period for both geese species can exceed the recognized hazardous doses for thiram, and to a lesser extent for tebuconazole, which indicates that some pesticides may be playing a previously overlooked role in the decline of red-breasted geese.

Phylogenetic Relationships of Avian Cestodes from Brine Shrimp and Congruence with Larval Morphology.

Determining molecular markers for parasites provides a useful tool for their identification, particularly for larval stages with few distinguishable diagnostic characters. Avian cestodes play a key role in the food webs and biodiversity of hypersaline wetlands, yet they remain understudied. Using naturally infected Artemia, we identified cestode larvae (cysticercoids), assessed their genetic diversity, and explored phylogenetic relationships in relation to larval morphology and waterbird final hosts. We obtained partial 18S rDNA sequences for 60 cysticercoids of the family Hymenolepidae infecting Artemia spp. from seven localities and three countries (Spain, the USA, and Chile). We present the first DNA sequences for six taxa: Confluaria podicipina, Fimbriarioides sp., Flamingolepis liguloides, Flamingolepis sp. 1, Flamingolepis sp. 2, and Hymenolepis californicus. Intraspecific sequence variation (0.00-0.19% diversity) was lower than intergroup genetic distance (0.7-14.75%). Phylogenetic analysis revealed three main clades: 1-Flamingolepis, 2-Fimbriarioides, 3-Confluaria and Hymenolepis, all of which separated from hymenolepidids from mammals and terrestrial birds. This clear separation among taxa is congruent with previous morphological identification, validating the 18S gene as a useful marker to discriminate at generic/species level. Working with intermediate hosts allows the expansion of knowledge of taxonomic and genetic diversity of cestodes in wildlife, as well as elucidation of their life cycles.

Connectivity at the human-wildlife interface: starling movements relate to carriage of E. coli.

Synanthropic bird species in human, poultry or livestock environments can increase the spread of pathogens and antibiotic-resistant bacteria between wild and domestic animals. We present the first telemetry-based spatial networks for a small songbird. We quantified landscape connectivity exerted by spotless starling movements, and aimed to determine if connectivity patterns were related to carriage of potential pathogens. We captured 28 starlings on a partridge farm in 2020 and tested them for Avian influenza virus, West Nile virus WNV, Avian orthoavulavirus 1, Coronavirus, Salmonella spp. and Escherichia coli. We did not detect any viruses or Salmonella, but one individual had antibodies against WNV or cross-reacting Flaviviruses. We found E. coli in 61 % (17 of 28) of starlings, 76 % (13 of 17) of which were resistant to gentamicin, 12 % (2 of 17) to cefotaxime/enrofloxacin and 6 % (1 of 17) were phenotypic extended spectrum beta-lactamase (ESBL) carriers. We GPS-tracked 17 starlings and constructed spatial networks showing how their movements (i.e. links) connect different farms with nearby urban and natural habitats (i.e. nodes with different attributes). Using E. coli carriage as a proxy for acquisition/dispersal of bacteria, we found differences across spatial networks constructed for E. coli positive (n = 7) and E. coli negative (n = 9) starlings. We used Exponential Random Graph Models to reveal significant differences between networks. In particular, an urban roost was more connected to other sites by movements of E. coli positive than by movements of E. coli negative starlings. Furthermore, an open pine forest used mainly for roosting was more connected to other sites by movements of E. coli negative than by movements of E. coli positive starlings. Using E. coli as a proxy for a potential pathogen carried by starlings, we reveal the pathways of spread that starlings could provide between farms, urban and natural habitats.

Demographic and conservation genomic assessment of the threatened marbled teal (Marmaronetta angustirostris).

Genetic assessment of species that have experienced dramatic population declines provides critical information that is instrumental for the design of conservation recovery programs. Here, we use different sources of molecular data (mtDNA and ddRAD-seq) to evaluate the genetic status of wild and captive populations of marbled teal (Marmaronetta angustirostris), a duck species classified as critically endangered in Spain and near threatened at a global scale. First, we determined the evolutionary and demographic trajectories of the wild population from Spain and the currently much larger population from Iraq, which is also the documented source of European zoo stocks. Second, we evaluated the suitability of the different captive populations for ongoing restocking programs in Spain and assessed their potential impact on the genetic composition of wild populations. Populations from Spain and Iraq were assigned to distinct genetic clusters, albeit with an overall low level of genetic differentiation, in line with their recent divergence (<8000 years ago) and lack of phylogeographic structure in the species. Demogenomic inferences revealed that the two populations have experienced parallel demographic trajectories, with a marked bottleneck during the last glacial period followed by a sudden demographic expansion and stability since the onset of the Holocene. The wild population from Spain presented high levels of inbreeding, but we found no evidence of recent genetic bottlenecks compatible with the human-driven decline of the species during the past century. The captive populations from the two Spanish centers involved in restocking programs showed genetic introgression from European zoos; however, we found limited evidence of introgression from the zoo genetic stock into the wild population from Spain, suggesting captive-bred birds have limited breeding success in the wild. Our study illustrates how ex situ conservation programs should consider the genetic distinctiveness of populations when establishing breeding stocks and highlights the importance of genetically assessing captive populations prior to reinforcement actions.

Variability of faecal microbiota and antibiotic resistance genes in flocks of migratory gulls and comparison with the surrounding environment.

Gulls commonly rely on human-generated waste as their primary food source, contributing to the spread of antibiotic-resistant bacteria and their resistance genes, both locally and globally. Our understanding of this process remains incomplete, particularly in relation to its potential interaction with surrounding soil and water. We studied the lesser black-backed gull, Larus fuscus, as a model to examine the spatial variation of faecal bacterial communities, antibiotic resistance genes (ARGs), and mobile genetic elements (MGEs) and its relationship with the surrounding water and soil. We conducted sampling campaigns within a connectivity network of different flocks of gulls moving across functional units (FUs), each of which represents a module of highly interconnected patches of habitats used for roosting and feeding. The FUs vary in habitat use, with some gulls using more polluted sites (notably landfills), while others preferring more natural environments (e.g., wetlands or beaches). Faecal bacterial communities in gulls from flocks that visit and spend more time in landfills exhibited higher richness and diversity. The faecal microbiota showed a high compositional overlap with bacterial communities in soil. The overlap was greater when compared to landfill (11%) than to wetland soils (6%), and much lower when compared to bacterial communities in surrounding water (2% and 1% for landfill and wetland water, respectively). The relative abundance of ARGs and MGEs were similar between FUs, with variations observed only for specific families of ARGs and MGEs. When exploring the faecal carriage of ARGs and MGEs in bird faeces relative to soil and water compartments, gull faeces were enriched in ARGs classified as High-Risk. Our results shed light on the complex dynamics of antibiotic resistance spread in wild bird populations, providing insights into the interactions among gull movement and feeding behavior, habitat characteristics, and the dissemination of antibiotic resistance determinants across environmental reservoirs.

Leakage of plastics and other debris from landfills to a highly protected lake by wintering gulls.

GENERAL CONTEXT: Gulls ingest plastic and other litter while foraging in open landfills, because organic matter is mixed with other debris. Therefore, gulls are potential biovectors of plastic pollution into natural habitats, especially when they concentrate in wetlands for roosting. NOVELTY: We quantified, for the first time, the flow of plastic and other anthropogenic debris from open landfills to a natural lake via the movement of gulls. We focused on Fuente de Piedra, an inland closed-basin lake in Spain that is internationally important for biodiversity. METHODOLOGY: In 2022, we sampled gull pellets regurgitated in the lake by lesser black-backed gulls Larus fuscus that feed on landfills, as well as their faeces, then characterized and quantified debris particles of ≥0.5 mm. By combining GPS and census data from 2010 to 2022, together with plastic quantification based on FTIR-ATR analysis, we estimated the average annual deposition of plastic and other debris by the wintering gull population into the lake. MAIN RESULTS: 86 % of pellets contained plastics, and 94 % contained other debris such as glass and textiles. Polyethylene (54 %), polypropylene (11.5 %) and polystyrene (11.5 %) were the main plastic polymers. An estimated annual mean of 400 kg of plastics were moved by gulls into the lake. Only 1 % of plastic mass was imported in faeces. DISCUSSION: Incorporating the biovectoring role of birds can provide a more holistic view of the plastic cycle and waste management. Biovectoring is predictable in sites worldwide where gulls and other waterbirds feed in landfills and roost in wetlands. We discuss bird deterrence and other ways of mitigating debris leakage into aquatic ecosystems.

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.

High prevalence of cestodes in Artemia spp. throughout the annual cycle: relationship with abundance of avian final hosts.

Brine shrimp, Artemia spp., act as intermediate hosts for a range of cestode species that use waterbirds as their final hosts. These parasites can have marked influences on shrimp behavior and fecundity, generating the potential for cascading effects in hypersaline food webs. We present the first comprehensive study of the temporal dynamics of cestode parasites in natural populations of brine shrimp throughout the annual cycle. Over a 12-month period, clonal Artemia parthenogenetica were sampled in the Odiel marshes in Huelva, and the sexual Artemia salina was sampled in the Salinas de Cerrillos in Almería. Throughout the year, 4-45 % of A. parthenogenetica were infected with cestodes (mean species richness = 0.26), compared to 27-72 % of A. salina (mean species richness = 0.64). Ten cestode species were recorded. Male and female A. salina showed similar levels of parasitism. The most prevalent and abundant cestodes were those infecting the most abundant final hosts, especially the Greater Flamingo Phoenicopterus ruber. In particular, the flamingo parasite Flamingolepis liguloides had a prevalence of up to 43 % in A. parthenogenetica and 63.5 % in A. salina in a given month. Although there was strong seasonal variation in prevalence, abundance, and intensity of cestode infections, seasonal changes in bird counts were weak predictors of the dynamics of cestode infections. However, infection levels of Confluaria podicipina in A. parthenogenetica were positively correlated with the number of their black-necked grebe Podiceps nigricollis hosts. Similarly, infection levels of Anomotaenia tringae and Anomotaenia microphallos in A. salina were correlated with the number of shorebird hosts present the month before. Correlated seasonal transmission structured the cestode community, leading to more multiple infections than expected by chance.

Long term impacts of endozoochory and salinity on germination of wetland plants after entering simulated seed banks.

Migratory waterbirds disperse a broad range of angiosperms by endozoochory (seed dispersal via gut passage), especially plants in coastal wetlands. However, there is no previous information about the capacity of seeds to remain in the seed bank after waterbird endozoochory, and very little about how wetland salinity can influence the effect of gut passage on germination. We collected seeds of Juncus subulatus (Juncaceae), Bolboschoenus maritimus, and Schoenoplectus litoralis (Cyperaceae) from Doñana marshes in Spain. All three species are considered to have physiological dormancy. After gut passage following ingestion by ducks, seeds were stored in darkness in solutions with six different conductivities (1, 2, 4, 8, 16, and 32 dSm-1), for periods of 1, 6, or 12 months to simulate presence in a seed bank. After storage, 1800 seeds of each plant species assigned to these treatments were subjected to germination tests in demineralized water, together with 1800 control seeds that had not been ingested before storage. All species germinated readily after storage, with or without gut passage beforehand. Storage time and salinity both had important effects on germinability and time to germination, which differed between control and ingested seeds, and between plant species. After ≥6 months, germinability of Cyperaceae was enhanced by gut passage (≤25% higher than control seeds) at some salinities. Only J. subulatus showed consistently lower germinability after passage (≤30%). Only B. maritimus showed consistently slower germination after passage (≤33%). Salinity effects were more complex after passage, but increasing salinity did not generally have a negative impact on germination of ingested seeds. When compared to additional seeds that had not been stored before germination tests, storage reduced germinability in J. subulatus (≤39% reduction), but increased it in B. maritimus (≤17%) and S. litoralis (≤46%). Seeds dispersed by waterbird endozoochory may be easily incorporated into wetland seed banks, where they can remain halotolerant and delay germination until conditions become suitable. This can benefit wetland plants by increasing rates of long-distance dispersal, gene flow, and establishment of new populations. Avian gut passage can have positive and species-specific effects on germination in plants with persistent seed banks and/or physiological dormancy.