Concurrent invasions of European starlings in Australia and North America reveal population-specific differentiation in shared genomic regions.

A species' success during the invasion of new areas hinges on an interplay between the demographic processes common to invasions and the specific ecological context of the novel environment. Evolutionary genetic studies of invasive species can investigate how genetic bottlenecks and ecological conditions shape genetic variation in invasions, and our study pairs two invasive populations that are hypothesized to be from the same source population to compare how each population evolved during and after introduction. Invasive European starlings (Sturnus vulgaris) established populations in both Australia and North America in the 19th century. Here, we compare whole-genome sequences among native and independently introduced European starling populations to determine how demographic processes interact with rapid evolution to generate similar genetic patterns in these recent and replicated invasions. Demographic models indicate that both invasive populations experienced genetic bottlenecks as expected based on invasion history, and we find that specific genomic regions have differentiated even on this short evolutionary timescale. Despite genetic bottlenecks, we suggest that genetic drift alone cannot explain differentiation in at least two of these regions. The demographic boom intrinsic to many invasions as well as potential inversions may have led to high population-specific differentiation, although the patterns of genetic variation are also consistent with the hypothesis that this infamous and highly mobile invader adapted to novel selection (e.g., extrinsic factors). We use targeted sampling of replicated invasions to identify and evaluate support for multiple, interacting evolutionary mechanisms that lead to differentiation during the invasion process.

Environmental correlates of genetic variation in the invasive European starling in North America.

Populations of invasive species that colonize and spread in novel environments may differentiate both through demographic processes and local selection. European starlings (Sturnus vulgaris) were introduced to New York in 1890 and subsequently spread throughout North America, becoming one of the most widespread and numerous bird species on the continent. Genome-wide comparisons across starling individuals and populations can identify demographic and/or selective factors that facilitated this rapid and successful expansion. We investigated patterns of genomic diversity and differentiation using reduced-representation genome sequencing of 17 winter-season sampling sites. Consistent with this species' high dispersal rate and rapid expansion history, we found low geographical differentiation and few FST outliers even at a continental scale. Despite starting from a founding population of ~180 individuals, North American starlings show only a moderate genetic bottleneck, and models suggest a dramatic increase in effective population size since introduction. In genotype-environment associations we found that ~200 single-nucleotide polymorphisms are correlated with temperature and/or precipitation against a background of negligible genome- and range-wide divergence. Given this evidence, we suggest that local adaptation in North American starlings may have evolved rapidly even in this wide-ranging and evolutionarily young system. This survey of genomic signatures of expansion in North American starlings is the most comprehensive to date and complements ongoing studies of world-wide local adaptation in these highly dispersive and invasive birds.

Nutritional depletion of total mixed rations by red-winged blackbirds and projected impacts on dairy cow performance.

This Research Communication describes an investigation of the nutritional depletion of total mixed rations (TMR) by pest birds. We hypothesized that species-specific bird depredation of TMR can alter the nutritional composition of the ration and that these changes can negatively impact the performance of dairy cows. Blackbirds selected the high energy fraction of the TMR (i.e., flaked corn) and reduced starch, crude fat and total digestible nutrients during controlled feeding experiments. For Holsteins producing 37·1 kg of milk/d, dairy production modeling illustrated that total required net energy intake (NEI) was 35·8 Mcal/d. For the reference TMR unexposed to blackbirds and the blackbird-consumed TMR, NEI supplied was 41·2 and 37·8 Mcal/d, and the resulting energy balance was 5·4 and 2·0 Mcal/d, respectively. Thus, Holsteins fed the reference and blackbird-consumed TMR were estimated to gain one body condition score in 96 and 254 d, and experience daily weight change due to reserves of 1·1 and 0·4 kg/d, respectively. We discuss these results in context of an integrated pest management program for mitigating the depredation caused by pest birds at commercial dairies.

Recent beak evolution in North American starlings after invasion.

European starlings are one of the most abundant and problematic avian invaders in the world. From their native range across Eurasia and North Africa, they have been introduced to every continent except Antarctica. In 160 years, starlings have expanded into different environments throughout the world, making them a powerful model for understanding rapid evolutionary change and adaptive plasticity. Here, we investigate their spatiotemporal morphological variation in North America and the native range. Our dataset includes 1217 specimens; a combination of historical museum skins and modern birds. Beak length in the native range has remained unchanged during the past 206 years, but we find beak length in North American birds is now 8% longer than birds from the native range. We discuss potential drivers of this pattern including dietary adaptation or climatic pressures. Additionally, body size in North American starlings is smaller than those from the native range, which suggests a role for selection or founder effect. Taken together, our results indicate rapid recent evolutionary change in starling morphology coincident with invasion into novel environments.

Transcript- and annotation-guided genome assembly of the European starling.

The European starling, Sturnus vulgaris, is an ecologically significant, globally invasive avian species that is also suffering from a major decline in its native range. Here, we present the genome assembly and long-read transcriptome of an Australian-sourced European starling (S. vulgaris vAU), and a second, North American, short-read genome assembly (S. vulgaris vNA), as complementary reference genomes for population genetic and evolutionary characterization. S. vulgaris vAU combined 10× genomics linked-reads, low-coverage Nanopore sequencing, and PacBio Iso-Seq full-length transcript scaffolding to generate a 1050 Mb assembly on 6222 scaffolds (7.6 Mb scaffold N50, 94.6% busco completeness). Further scaffolding against the high-quality zebra finch (Taeniopygia guttata) genome assigned 98.6% of the assembly to 32 putative nuclear chromosome scaffolds. Species-specific transcript mapping and gene annotation revealed good gene-level assembly and high functional completeness. Using S. vulgaris vAU, we demonstrate how the multifunctional use of PacBio Iso-Seq transcript data and complementary homology-based annotation of sequential assembly steps (assessed using a new tool, saaga) can be used to assess, inform, and validate assembly workflow decisions. We also highlight some counterintuitive behaviour in traditional busco metrics, and present buscomp, a complementary tool for assembly comparison designed to be robust to differences in assembly size and base-calling quality. This work expands our knowledge of avian genomes and the available toolkit for assessing and improving genome quality. The new genomic resources presented will facilitate further global genomic and transcriptomic analysis on this ecologically important species.

Toxicity of sodium nitrite-based vertebrate pesticides for European starlings (Sturnus vulgaris).

In the 21st century, invasive animals rank second only to habitat destruction as the greatest threat to global biodiversity. Socially-acceptable and cost-effective strategies are needed to reduce the negative economic and environmental impacts of invasive animals. We investigated the potential for sodium nitrite (SN; CAS 7632-00-0) to serve as an avian toxicant for European starlings (Sturnus vulgaris L.). We also assessed the non-target hazard of an experimental formulation of SN that is being developed as a toxicant for invasive wild pigs (Sus scrofa L.). In gavage experiments with European starlings, we identified a lowest observed adverse effect level (LOAEL) for mortality of 2.40% technical SN (w/v; 120 mg SN/kg body mass) and a no observed adverse effect level (NOAEL) for mortality of 1.30% technical SN (65 mg/kg). The exposure of ten starlings to the experimental formulation of SN (10% SN pig toxicant) resulted in one starling mortality during four days of exposure to the toxic bait. Sodium nitrite toxicity presented a moderate hazard to European starlings; thus, the future development of SN as an avian toxicant is dependent upon its cost-effectiveness. We discuss the management of toxic effects and non-target hazards of SN for wild birds, including best practices for toxic baiting of vertebrate pests and management of invasive wild pigs.

Inefficiency of anthraquinone-based avian repellents when applied to sunflower: the importance of crop vegetative and floral characteristics in field applications.

BACKGROUND: Blackbirds (Icteridae) cause significant damage to sunflower (Helianthus annuus L.) prompting the need for effective management tools. Anthraquinone-based repellents can reduce feeding by > 80% in laboratory settings, but require birds to learn the negative association through repellent ingestion. We evaluated an anthraquinone-based repellent applied directly to mature sunflower plants for its ability to reduce bird damage. We used captive male red-winged blackbirds (Agelaius phoeniceus) to evaluate efficacy of two anthraquinone-based formulations in varying concentrations and applied in a manner attainable by sunflower producers. We also assessed field application methods for repellent coverage and anthraquinone residues when using ground-rigs equipped with drop-nozzles situated below the crop canopy. RESULTS: The repellents failed to reduce feeding and birds did not exhibit a preference between untreated and treated sunflowers at concentrations 2.7× the suggested application rate (i.e. 9.35 L ha-1 of repellent). In the absence of disk flowers, which obstruct repellent from reaching the achenes, the repellents failed to reduce consumption. Anthraquinone concentrations in field applications were considerably less than those in the laboratory experiments and did not reduce bird damage. CONCLUSION: Efficacy is difficult to achieve in the field due to application issues where growth patterns and floral components of sunflower limit residues on achenes, thus contact with foraging birds. Although field residues could be improved by increasing anthraquinone concentrations in tank mixtures and decreasing droplet size, repellents optimized for loose achenes are inefficient in reducing avian consumption of sunflower when applied to intact plants in a manner representative of commercial agriculture. © 2020 Society of Chemical Industry.

Multi-isotopic (δ2H, δ13C, δ15N) tracing of molt origin for European starlings associated with U.S. dairies and feedlots.

Introduced bird species can become invasive in agroecosystems and their management is inhibited if their origin and movements are not well understood. Stable isotope measurements of feathers can be used to infer molt origins and interstate movements in North America. We analyzed stable-hydrogen (δ2H), carbon (δ13C) and nitrogen (δ 15N) isotope ratios in feathers to better understand the molt origin of European starlings (Sturnus vulgaris) collected at dairies and feedlots throughout the United States. Primary feathers were used from 596 adult and 90 juvenile starlings collected during winter at dairies and feedlots that experience starling damages in Arizona, California, Colorado, Idaho, Illinois, Iowa, Kansas, Minnesota, Missouri, Nebraska, Nevada, New Hampshire, New Mexico, New York, North Carolina, Oregon, Texas, Vermont, Washington and Wisconsin. The best-fit model indicated that the combination of feather δ2H, δ13C and δ15N values best predicted the state where samples were collected and thus supported use of this approach for tracing molt origins in European starlings. Interestingly, molt origins of starlings collected at dairies and feedlots generally west of -90° longitude (i.e. 11 of 15 states west of the Mississippi River, including Wisconsin) were assigned to the collection state and/or the state adjacent to the collection state. In contrast, molt origins of starlings collected generally east of -90° longitude (four of five eastern states) were not assigned to the collection state and/or the state adjacent to the collection state. Among all starlings (N = 686), 23% were assigned to the collection state and 19% were assigned to the state adjacent to the collection state. Among all males (N = 489) and all females (N = 197), 23% and 26% were assigned to the collection state and 19% and 13% were assigned to the state adjacent to the collection state, respectively. We observed a greater proportion (88%) of juvenile starlings assigned to states other than their collection state (i.e. potentially a result of natal dispersal) than that proportion (76%) in adult starlings. This study included an unprecedented sample of feather isotopes from European starlings throughout the United States. As a novel contribution to the ecology and management of invasive and migratory passerines, we demonstrate how such feather isoscapes can be used to predict molt origin and, potentially, interstate movements of European starlings for subsequent ecological and management investigations.

Food color, flavor, and conditioned avoidance among red-winged blackbirds.

The relationship between food flavors and postingestive feedback enables mammalian herbivores to procure nutrients and avoid toxins within ever-changing environments. We conducted four experiments with red-winged blackbirds (Agelaius phoeniceus) in captivity to determine the relative roles of color and flavor cues paired with negative postingestive feedback. We first conducted baseline tests to assess preferences for colors and flavors. All blackbirds preferred red- to blue-colored food, and they preferred umami- (l-alanine) flavored to bitter/astringent food (tannic acid). We observed no difference in consumption of salty (NaCl) vs sour (citric acid) foods during baseline tests (i.e., neutral flavors). We then conditioned experimentally naïve blackbirds with intraperitoneal injections of lithium chloride (LiCl) to avoid food treated with red and l-alanine, or red and NaCl (n=30 birds per conditioning group). Subsequent to conditioning with LiCl, three test groups were established from each conditioned group to evaluate color and flavor preferences, and preferences for novel color-flavor pairings (e.g., red/tannic acid vs blue/l-alanine). Blackbirds avoided red and salty food throughout the 4-day test. Avoidance conditioned with LiCl extinguished for preferred flavors, but not for colors, of food. Conditioning affected indifference for the otherwise preferred flavor and avoidance for the otherwise neutral flavor. Relative to the neutral-flavor conditioning group, the group conditioned with a preferred flavor exhibited stronger conditioned avoidance of colored food. Unlike conditioned flavor avoidance, birds were conditioned to avoid red food only when blue food was made familiar prior to conditioning. Collectively, these results illustrate that blackbirds used affective processes (flavor-feedback relationships) to shift preference for both novel and familiar flavors, and cognitive associations (colors) to avoid food, subsequent to toxin exposure. We discuss the opportunities afforded by affective and cognitive processing for reducing agricultural damage caused by blackbirds.

Bird use of organic apple orchards: Frugivory, pest control and implications for production.

As the largest terrestrial biomes, crop and pasturelands can have very large positive or negative impacts on biodiversity and human well-being. Understanding how animals use and impact agroecosystems is important for making informed decisions that achieve conservation and production outcomes. Yet, few studies examine the tradeoffs associated with wildlife in agricultural systems. We examined bird use of organic apple orchards as well as how birds influence fruit production positively through control of an economically important insect pest (codling moth (Cydia pomonella)) and negatively through fruit damage. We conducted transect surveys, observed bird frugivory and assessed bird and insect damage with an exclosure experiment in small organic farms in western Colorado. We found that organic apple orchards in this region provide habitat for a large number of both human-adapted and human-sensitive species and that the species in orchards were relatively similar to adjacent hedgerow habitats. Habitat use did not vary as a function of orchard characteristics, and apple damage by both birds and C. pomonella was consistent within and across apple blocks that varied in size. A small subset of bird species was observed foraging on apples yet the effect of birds as agents of fruit damage appeared rather minor and birds did not reduce C. pomonella damage. Our results demonstrate that organic apple orchards have the potential to provide habitat for diverse bird communities, including species typically sensitive to human activities, with little apparent effect on production.

Functional visual sensitivity to ultraviolet wavelengths in the Pileated Woodpecker (Dryocopus pileatus), and its influence on foraging substrate selection.

Most diurnal birds are presumed visually sensitive to near ultraviolet (UV) wavelengths, however, controlled behavioral studies investigating UV sensitivity remain few. Although woodpeckers are important as primary cavity excavators and nuisance animals, published work on their visual systems is limited. We developed a novel foraging-based behavioral assay designed to test UV sensitivity in the Pileated Woodpecker (Dryocopus pileatus). We acclimated 21 wild-caught woodpeckers to foraging for frozen mealworms within 1.2m sections of peeled cedar (Thuja spp.) poles. We then tested the functional significance of UV cues by placing frozen mealworms behind UV-reflective covers, UV-absorptive covers, or decayed red pine substrates within the same 1.2m poles in independent experiments. Behavioral responses were greater toward both UV-reflective and UV-absorptive substrates in three experiments. Study subjects therefore reliably differentiated and attended to two distinct UV conditions of a foraging substrate. Cue-naïve subjects showed a preference for UV-absorptive substrates, suggesting that woodpeckers may be pre-disposed to foraging from such substrates. Behavioral responses were greater toward decayed pine substrates (UV-reflective) than sound pine substrates suggesting that decayed pine can be a useful foraging cue. The finding that cue-naïve subjects selected UV-absorbing foraging substrates has implications for ecological interactions of woodpeckers with fungi. Woodpeckers transport fungal spores, and communication methods analogous to those of plant-pollinator mutualisms (i.e. UV-absorbing patterns) may have evolved to support woodpecker-fungus mutualisms.

Review of anthraquinone applications for pest management and agricultural crop protection.

We have reviewed published anthraquinone applications for international pest management and agricultural crop protection from 1943 to 2016. Anthraquinone (AQ) is commonly found in dyes, pigments and many plants and organisms. Avian repellent research with AQ began in the 1940s. In the context of pest management, AQ is currently used as a chemical repellent, perch deterrent, insecticide and feeding deterrent in many wild birds, and in some mammals, insects and fishes. Criteria for evaluation of effective chemical repellents include efficacy, potential for wildlife hazards, phytotoxicity and environmental persistence. As a biopesticide, AQ often meets these criteria of efficacy for the non-lethal management of agricultural depredation caused by wildlife. We summarize published applications of AQ for the protection of newly planted and maturing crops from pest birds. Conventional applications of AQ-based repellents include preplant seed treatments [e.g. corn (Zea mays L.), rice (Oryza sativa L.), sunflower (Helianthus annuus L.), wheat (Triticum spp.), millet (Panicum spp.), sorghum (Sorghum bicolor L.), pelletized feed and forest tree species] and foliar applications for rice, sunflower, lettuce (Lactuca sativa L.), turf, sugar beets (Beta vulgaris L.), soybean (Glycine max L.), sweet corn and nursery, fruit and nut crops. In addition to agricultural repellent applications, AQ has also been used to treat toxicants for the protection of non-target birds. Few studies have demonstrated AQ repellency in mammals, including wild boar (Sus scrofa, L.), thirteen-lined ground squirrels (Ictidomys tridecemlineatus, Mitchill), black-tailed prairie dogs (Cyomys ludovicainus, Ord.), common voles (Microtus arvalis, Pallas), house mice (Mus musculus, L.), Tristram's jirds (Meriones tristrami, Thomas) and black rats (Rattus rattus L.). Natural sources of AQ and its derivatives have also been identified as insecticides and insect repellents. As a natural or synthetic biopesticide, AQ is a promising candidate for many contexts of non-lethal and insecticidal pest management. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

Multi-Isotopic (δ2H, δ13C, δ15N) Tracing of Molt Origin for Red-Winged Blackbirds Associated with Agro-Ecosystems.

We analyzed stable-hydrogen (δ2H), carbon (δ13C) and nitrogen (δ 15N) isotope ratios in feathers to better understand the molt origin and food habits of Red-winged Blackbirds (Agelaius phoeniceus) near sunflower production in the Upper Midwest and rice production in the Mid-South of the United States. Outer primary feathers were used from 661 after-second-year (ASY) male blackbirds collected in Minnesota, Montana, North Dakota and South Dakota (spring collection), and Arkansas, Louisiana, Mississippi, Missouri and Texas (winter collection). The best-fit model indicated that the combination of feather δ2H, δ13C and δ15N best predicted the state of sample collections and thus supported the use of this approach for tracing molt origins in Red-winged Blackbirds. When considering only birds collected in spring, 56% of birds were classified to their collection state on the basis of δ2H and δ13C alone. We then developed feather isoscapes for δ13C based upon these data and for δ2H based upon continental patterns of δ2H in precipitation. We used 81 birds collected at the ten independent sites for model validation. The spatially-explicit assignment of these 81 birds to the δ2H isoscape resulted in relatively high rates (~77%) of accurate assignment to collection states. We also modeled the spatial extent of C3 (e.g. rice, sunflower) and C4 (corn, millet, sorghum) agricultural crops grown throughout the Upper Midwest and Mid-South United States to predict the relative use of C3- versus C4-based foodwebs among sampled blackbirds. Estimates of C3 inputs to diet ranged from 50% in Arkansas to 27% in Minnesota. As a novel contribution to blackbird conservation and management, we demonstrate how such feather isoscapes can be used to predict the molt origin and interstate movements of migratory blackbirds for subsequent investigations of breeding biology (e.g. sex-specific philopatry), agricultural depredation, feeding ecology, physiology of migration and sensitivity to environmental change.

Proximal cues of pocket gopher burrow plugging behavior: influence of light, burrow openings, and temperature.

Burrow plugging is readily observed among mammals adapted for digging (i.e., fossorial mammals) as they create and maintain their burrows. We investigated the influence of light, burrow openings, and thermal environment as cues of pocket gopher (Thomomys mazama, Thomomys talpoides) behavior. When given free access to light and no light during artificial-burrow preference trials, both Thomomys spp. consistently plugged (i.e., avoided) light treatments. Burrow openings did not notably affect plugging behavior of T. mazama. Gophers (T. talpoides) plugged the artificial burrows within the light and cold (7 degrees C) treatments, but not within the no-light, and 18 or 31 degrees C treatments when light and temperature were varied independently. Whereas the presence of light and low ambient temperatures induce burrow maintenance by pocket gophers, these cues help meliorate adverse conditions within subsurface environs.

Functional significance of ultraviolet feeding cues in wild turkeys.

Most birds are able to sense ultraviolet (UV) visual signals. Ultraviolet wavelengths are used for plumage signaling and sexual selection among birds. The aim of our study was to determine if UV cues are also used for the process of food selection in wild turkeys (Meleagris gallopavo). We used avoidance conditioning to test the hypothesis that UV feeding cues can be used functionally for foraging behavior in wild turkeys. Female turkeys exhibited no avoidance of untreated food and 75­98% avoidance of food treated with an UV-absorbent, postingestive repellent (0.5­4% anthraquinone; wt./wt.) during repellent exposure. Male turkeys exhibited 78­99% avoidance of food treated with 0.5­4% anthraquinone. Female and male turkeys that consumed more than 200 mg and 100 mg of anthraquinone, respectively, subsequently avoided food treated only with an UV-absorbent cue. In contrast, unconditioned females consumed 58% more food treated with the UV-absorbent cue than untreated food. Thus, wild turkeys do not prefer foods associated with UV wavelengths regardless of feeding experience. We also observed 1) a weak negative correlation between body condition and intestinal parasite infection and 2) moderate, positive correlations between consumption of food treated with the conditioned UV cue and intestinal parasite infection among male turkeys. The UV feeding cue was used to maintain food avoidance during the four days subsequent to postingestive conditioning. Moreover, the consequences of consuming food treated with the postingestive, UV-absorbent repellent were necessary for conditioned avoidance of the UV-absorbent cue. These findings suggest functional significance of UV feeding cues for avian foraging behavior, the implications of which will enable subsequent investigations regarding the sensory physiology and behavioral ecology of wild birds.

The role of a generalized ultraviolet cue for blackbird food selection.

Birds utilize ultraviolet (UV) wavelengths for plumage signaling and sexual selection. Ultraviolet cues may also be used for the process of avian food selection. The aim of our study was to investigate whether a UV cue and a postingestive repellent can be used to condition food avoidance in red-winged blackbirds (Agelaius phoeniceus). We found that birds conditioned with an UV-absorbent, postingestive repellent subsequently avoided UV-absorbent food. Thus, the UV-absorbent cue (coupled with 0-20% of the conditioned repellent concentration) was used to maintain avoidance for up to 18 days post-conditioning. Similarly, birds conditioned with the UV-absorbent, postingestive repellent subsequently avoided UV-reflective food. Thus, conditioned avoidance of an UV-absorbent cue can be generalized to an unconditioned, UV-reflective cue for nutrient selection and toxin avoidance. These findings support the hypothesized function of UV vision for avian food selection, the implications of which remain to be explored for the sensory and behavioral ecology within agronomic and natural environments.

Reconciling sensory cues and varied consequences of avian repellents.

We learned previously that red-winged blackbirds (Agelaius phoeniceus) use affective processes to shift flavor preference, and cognitive associations (colors) to avoid food, subsequent to avoidance conditioning. We conducted three experiments with captive red-winged blackbirds to reconcile varied consequences of treated food with conditioned sensory cues. In Experiment 1, we compared food avoidance conditioned with lithium chloride (LiCl) or naloxone hydrochloride (NHCl) to evaluate cue-consequence specificity. All blackbirds conditioned with LiCl (gastrointestinal toxin) avoided the color (red) and flavor (NaCl) of food experienced during conditioning; birds conditioned with NHCl (opioid antagonist) avoided only the color (not the flavor) of food subsequent to conditioning. In Experiment 2, we conditioned experimentally naïve blackbirds using free choice of colored (red) and flavored (NaCl) food paired with an anthraquinone- (postingestive, cathartic purgative), methiocarb- (postingestive, cholinesterase inhibitor), or methyl anthranilate-based repellent (preingestive, trigeminal irritant). Birds conditioned with the postingestive repellents avoided the color and flavor of foods experienced during conditioning; methyl anthranilate conditioned only color (not flavor) avoidance. In Experiment 3, we used a third group of blackbirds to evaluate effects of novel comparison cues (blue, citric acid) subsequent to conditioning with red and NaCl paired with anthraquinone or methiocarb. Birds conditioned with the postingestive repellents did not avoid conditioned color or flavor cues when novel comparison cues were presented during the test. Thus, blackbirds cognitively associate pre- and postingestive consequences with visual cues, and reliably integrate visual and gustatory experience with postingestive consequences to procure nutrients and avoid toxins.

Dispersal of viable row-crop seeds of commercial agriculture by farmland birds: implication for genetically modified crops.

To address some concerns about the expansion of genetically engineered pharmaceutical and industrial crops to outdoor plantings and potential impacts on the human food supply, we determined whether commercial agriculture seeds of maize or corn Zea mays L., barley Hordeum vulgare L., safflower Carthamus tinctorius L. and rice Oryza sativa L. are digested or pass viably through the digestive tract, or are transported externally, by captive mallard ducks Anas platyrhynchos L., ring-necked pheasants Phasianus colchicus L., red-winged blackbirds Agelaius phoeniceus (L.) and rock pigeons Columba livia Gmelin (with the exception of whole maize seeds which were too large to feed to the blackbirds). These crop seeds, whether free-fed or force-fed, did not pass through the digestive tract of these bird species. The birds nonetheless did retain viable seeds in the esophagus/crop and gizzard for several hours. For example, after foraging for 6 h, mallards had retained an average of 228 +/- 112 barley seeds and pheasants 192 +/- 78 in the esophagus/crop, and their germination rates were 93 and 50%, respectively. Birds externally transported seeds away from the feeding location, but in only four instances were seeds found attached to their muddy feet or legs and in no case to feathers. Risk of such crop seeds germinating, establishing and reproducing off site after transport by a bird (externally or internally) or movement of a carcass by a predator, will depend greatly on the crop and bird species, location, environmental conditions (including soil characteristics), timing, and seed condition.