Predation of the endangered Ae'o (Hawaiian Stilt) by a native raptor, the Pueo (Hawaiian Short-eared Owl) on the island of O'ahu, Hawai'i, USA.

While the impact of introduced predators is a widely acknowledged issue and key component of conservation considerations for endemic waterbird populations in the Hawaiian Islands, the impact of native predators on endemic, endangered waterbirds is not as frequently discussed or factored into recovery models. The Pueo (Hawaiian Short-eared Owl; Asio flammeus sandwichensis) is a subspecies of Short-eared Owl endemic to the Hawaiian Islands and is State-listed as Endangered on the island of O'ahu. The Ae'o (Hawaiian Stilt; Himantopus mexicanus knudensi) is a subspecies of the Black-necked Stilt endemic to Hawai'i and is federally listed as Endangered throughout its range. A variety of non-native predators are confirmed to consume Ae'o eggs, chicks, and adults, including invasive mammals (e.g., feral cats), birds (e.g., Barn Owls), and amphibians (e.g., bullfrogs). While predation by native predators was suspected, there are no cases documented in the literature to date describing Pueo preying upon Ae'o. Here, we describe four events that provide evidence of Pueo predating Ae'o during the 2019-2021 breeding seasons in a wetland area on the island of O'ahu: (1) confirmed Pueo predating an Ae'o chick, (2) a suspected predation attempt of a Pueo chasing adult Ae'o, and (3) two suspected predation events based on (a) 10 adult-sized Ae'o carcasses and remains found near an active Pueo nest and (b) game camera photos of Pueo visiting two Ae'o nests. To our knowledge, these novel observations are the first published accounts of predator-prey interactions between these two subspecies.

Dietary effects on fitness in captive-reared Hawaiian tree snails.

The native terrestrial snail fauna of the Hawaiian Islands faces numerous threats that have led to severe range reductions, population declines, and extinction of species. With the continued declines of many wild populations, a crucial component of preserving Hawaiian terrestrial snail biodiversity is through captive rearing programs, like that implemented by the Hawai'i Department of Land and Natural Resources Snail Extinction Prevention Program. Rare and endangered tree snails in the family Achatinellidae, which feed on epiphytic microbial communities, are maintained in captivity with a diet that includes native vegetation brought in from nearby forests, as well as a cultured fungus originally isolated from native host trees. Recent mortality events in lab populations have been attributed to wild-gathered vegetation. These events have increased interest in developing a completely manufactured or cultured diet that would eliminate the need for exposure to wild-gathered plants. This study compared survival and egg production in Auriculella diaphana provided with lab-cultured fungus, and those provided with wild vegetation. We compared the number of eggs laid and number of deaths among three treatments: (1) wild collected vegetation only; (2) wild vegetation supplemented with laboratory-cultured fungus; and (3) laboratory cultured fungus only. Mortality did not significantly differ among treatments, but the number of eggs laid was significantly higher in snails provided wild vegetation and cultured fungus (F = 24.998; P < 0.001), compared with those provided with only wild vegetation (t = 1.88, P = 0.032) or only cultured fungus (t = 4.530, P = 0.004). Our results suggest: (1) the existing strain of cultured fungus alone is not sufficient to maintain captive-reared snail populations; (2) the additional energy or calcium provided by the cultured fungus appears to enhance egg reproduction in captive-reared populations; (3) the presence or absence of live vegetation influences snail behavior, including aestivation and egg laying. These results highlight the importance of ongoing research to culture additional species of fungi at a rate that could support captive-reared populations, as the diversity of fungi present in wild epiphytic microbial communities may be important for snail reproductive health.

Wedge-tailed Shearwater (Ardenna pacifica) nesting success in human-dominated coastal environments.

Many seabird populations are declining globally, but successful conservation efforts have led to population expansion of some species into human-dominated landscapes. Thus, there is an increased potential for direct human and seabird interactions for certain species in human-occupied areas, with nest-site characteristics potentially affecting the susceptibility of nests to human disturbance. We assessed the effect of human activity and nest-site characteristics on Wedge-tailed Shearwater (Ardenna pacifica, 'ua'u kani) nesting success at two breeding colonies, one with human exposure and one without, located in Kailua, O'ahu, Hawai'i. Human activity was measured by recording the frequency of people who entered a 5 m buffer around each nest. Nests were checked every two to three days to monitor nest success. The effect of human activity and nest-site characteristics on nesting success was determined using a variety of combinations of variables within binomial logistic regression models and AICc model selection. Nest-site characteristics among nests at both sites and human activity at the human-exposed site did not show a significant effect on nesting success. Our results suggest Wedge-tailed Shearwaters may experience some tolerance of human activity immediately around their nests-as long as burrow collapse does not occur. Given the small sample sizes and a single season of data collection, additional studies are needed to better understand the effect of human disturbance on Wedge-tailed Shearwaters. Infrastructure, such as fencing and signage, may be effective at reducing human-caused nest failure and may allow humans and disturbance-tolerant seabird species to coexist in shared coastal environments.

The global impact of wild pigs (Sus scrofa) on terrestrial biodiversity.

The International Union for the Conservation of Nature's (IUCN) Red List of Threatened Species is a comprehensive database of over 120,000 species and is a powerful tool to evaluate the threat of invasive species to global biodiversity. Several problematic species have gained global recognition due to comprehensive threat assessments quantifying the threat these species pose to biodiversity using large datasets like the IUCN Red List of Threatened Species. However, the global threat of wild pigs (Sus scrofa) to biodiversity is still poorly understood despite well-documented ecosystem level impacts. In this study, we utilized the IUCN Red List to quantify the impacts of this globally distributed species throughout its native and non-native range. Here we show that wild pigs threaten 672 taxa in 54 different countries across the globe. Most of these taxa are listed as critically endangered or endangered and 14 species have been driven to extinction as a direct result of impacts from wild pigs. Our results show that threats from wild pigs are pervasive across taxonomic groups and that island endemics and taxa throughout the non-native range of wild pigs are particularly vulnerable.

Perceived Barriers to the Use of Assisted Colonization for Climate Sensitive Species in the Hawaiian Islands.

Conservation actions to safeguard climate change vulnerable species may not be utilized due to a variety of perceived barriers. Assisted colonization, the intentional movement and release of an organism outside its historical range, is one tool available for species predicted to lose habitat under future climate change scenarios, particularly for single island or single mountain range endemic species. Despite the existence of policies that allow for this action, to date, assisted colonization has rarely been utilized for species of conservation concern in the Hawaiian Islands. Given the potential for climate driven biodiversity loss, the Hawaiian Islands are a prime location for the consideration of adaptation strategies. We used first-person interviews with conservation decision makers, managers, and scientists who work with endangered species in the Hawaiian Islands to identify perceived barriers to the use of assisted colonization. We found that assisted colonization was often not considered or utilized due to a lack of expertize with translocations; ecological risk and uncertainty, economic constraints, concerns regarding policies and permitting, concerns with public perception, and institutional resistance. Therefore, conservation planners may benefit from decision tools that integrate risk and uncertainty into decision models, and compare potential outcomes among conservation actions under consideration, including assisted colonization. Within a decision framework that addresses concerns, all conservation actions for climate sensitive species, including assisted colonization, may be considered in a timely manner.

Cooperative breeding behaviors in the Hawaiian Stilt (Himantopus mexicanus knudseni).

Cooperative breeding, which is commonly characterized by nonbreeding individuals that assist others with reproduction, is common in avian species. However, few accounts have been reported in Charadriiformes, particularly island-nesting species. We present incidental observations of cooperative breeding behaviors in the Hawaiian Stilt (Himantopus mexicanus knudseni), an endangered subspecies of the Black-necked Stilt (Himantopus mexicanus), during the 2012-2020 nesting seasons on the Hawaiian islands of O'ahu and Moloka'i. We describe two different behaviors that are indicative of cooperative breeding: (a) egg incubation by multiple adults; (b) helpers-at-the-nest, whereby juveniles delay dispersal and reproduction to assist parents and siblings with reproduction. These observations are the first published accounts of cooperative breeding in this subspecies and merit further investigation, as cooperative breeding may improve population viability of the endangered, endemic Hawaiian Stilt.

Evolutionary genomics of endangered Hawaiian tree snails (Achatinellidae: Achatinellinae) for conservation of adaptive capacity.

Phylogenomic studies can provide insights into speciation, adaptation, and extinction, while providing a roadmap for conservation. Hawaiian tree snails are a model system for an adaptive radiation facing an extinction crisis. In the last 5 years, nearly all populations of Hawaiian tree snails across the 30 remaining species in the subfamily Achatinellinae (Achatinellidae) have declined from hundreds or thousands in the wild down to undetectable levels. Nearly 100 species historically occurred across dramatic environmental gradients on five of the Hawaiian Islands, but habitat loss, overcollection, and predation by invasive species have decimated populations. As such, this system offers the opportunity to integrate efforts to conserve evolutionary potential into conservation planning for a rapidly declining subfamily. Here, we used genome-wide, restriction-site associated DNA sequencing (RADseq), along with mitochondrial genome reconstruction, to resolve evolutionary relationships to inform conservation efforts. Phylogenetic analysis of nearly 400k genome-wide SNPs from 59 populations and 25 species across six genera in the family Achatinellidae, was generally concordant with taxonomy, geography, and mtDNA with several notable exceptions; mtDNA was unable to resolve some deeper nodes (e.g., the monophyly of Achatinella), while SNP data did not resolve as many shallow nodes. Both phylogenetic and coalescent analysis revealed deep divergences between populations within Achatinella mustelina that were consistent with species-level differences. Given cryptic species-level divergence within populations that are geographically proximate, they are at higher risk of extirpation from invasive predators and climate change than previously assumed. This study clarifies evolutionary relationships within this model system for adaptive radiation, forming the basis for conservation strategies such as translocation, captive rearing, and hybridization trials to prevent the loss of capacity to adapt to rapidly changing environmental conditions.

Mammal-exclusion fencing improves the nesting success of an endangered native Hawaiian waterbird.

Invasive predator control is often critical to improving the nesting success of endangered birds, but methods of control vary in cost and effectiveness. Poison-baiting or trapping and removal are relatively low-cost, but may have secondary impacts on non-target species, and may not completely exclude mammals from nesting areas. Mammal-exclusion fencing has a substantial up-front cost, but due to cost savings over the lifetime of the structure and the complete exclusion of mammalian predators, this option is increasingly being utilized to protect threatened species such as ground-nesting seabirds. However, non-mammalian predators are not excluded by these fences and may continue to impact nesting success, particularly in cases where the fence is designed for the protection of waterbirds, open to an estuary or wetland on one side. Thus, there remains a research gap regarding the potential gains in waterbird nesting success from the implementation of mammal-exclusion fencing in estuarine systems. In this study, we compared the nesting success of endangered Hawaiian Stilts (Ae'o; Himantopus mexicanus knudseni) within a mammal-exclusion fence to that of breeding pairs in a nearby wetland where trapping was the sole means for removing invasive mammals. We predicted success would be greater for breeding pairs inside the exclusion fence and the hatchlings inside the enclosure would spend more time in the nesting area than hatchlings at the unfenced site. During a single breeding season following construction of a mammal-exclusion fence, we used motion-activated game cameras to monitor nests at two sites, one site with mammal-exclusion fencing and one site without. Clutch sizes and hatch rates were significantly greater at the fenced site than the unfenced site, but time spent by chicks in the nesting area did not differ between sites. These results add to the mounting body of evidence that demonstrates the effectiveness of mammal-exclusion fencing in protecting endangered birds and suggests it can aid endangered Hawaiian waterbirds toward recovery. These results also suggest that the single greatest predatory threat to the Hawaiian Stilt may be invasive mammals, despite a host of known non-mammalian predators including birds, crabs, turtles, and bullfrogs, as the complete exclusion of mammals resulted in significant gains in nesting success. As additional fences are built, future studies are necessary to compare nesting success among multiple sites and across multiple seasons to determine potential gains in fledging success and recruitment.

Seasonal patterns in nest survival of a subtropical wading bird, the Hawaiian Stilt (Himantopus mexicanus knudseni).

Nest survival is influenced by where and when birds decide to breed. For ground-nesting species, nest-site characteristics, such as vegetation height and proximity to water, may impact the likelihood of nest flooding or depredation. Further, habitat characteristics, and thus nest survival, may fluctuate across the breeding season. The Hawaiian Stilt ('Ae'o; Himantopus mexicanus knudseni) is an endangered Hawaiian waterbird that nests in wetlands across the Hawaiian Islands. In this study, we used observational surveys and nest cameras to examine the impact of nest-site characteristics and day of nesting season on nest survival of the Hawaiian Stilt. Early nests had a higher chance of survival than late nests. For most of the nesting season, taller vegetation was correlated with increased nest survival, while shorter vegetation was correlated with increased nest survival late in the nesting season. Seasonal patterns in nest survival may be due to changes in parental behavior or predator activity. Nest depredation was responsible for 55% of confirmed nest failures and introduced mammals were the primary nest predators. Our study is the first to examine seasonality in nest survival of Hawaiian Stilts and suggests that, despite longer nesting seasons and year-round occupation of wetlands, late nesters in subtropical regions may have lower nest survival than early nesters, similar to trends observed in temperate regions.

Unexpectedly high genetic diversity in a rare and endangered seabird in the Hawaiian Archipelago.

Seabirds in the order of Procellariiformes have one of the highest proportions of threatened species of any avian order. Species undergoing recovery may be predicted to have a genetic signature of a bottleneck, low genetic diversity, or higher rates of inbreeding. The Hawaiian Band-rumped Storm Petrel ('Ake'ake; Hydrobates castro), a long-lived philopatric seabird, suffered massive population declines resulting in its listing under the Endangered Species Act in 2016 as federally Endangered. We used high-throughput sequencing to assess patterns of genetic diversity and potential for inbreeding in remaining populations in the Hawaiian Islands. We compared a total of 24 individuals, including both historical and modern samples, collected from breeding colonies or downed individuals found on the islands of Kaua'i, O'ahu, Maui, and the Big Island of Hawai'i. Genetic analyses revealed little differentiation between breeding colonies on Kaua'i and the Big Island colonies. Although small sample sizes limit inferences regarding other island colonies, downed individuals from O'ahu and Maui did not assign to known breeding colonies, suggesting the existence of an additional distinct breeding population. The maintenance of genetic diversity in future generations is an important consideration for conservation management. This study provides a baseline of population structure for the remaining nesting colonies that could inform potential translocations of the Endangered H. castro.

A comparison of mitochondrial genomes from five species in three genera suggests polyphyly in the subfamily Achatinellinae (Gastropoda: Pulmonata: Stylommatophora: Achatinellidae).

We compare the complete mitochondrial genomes of Achatinella fulgens, A. mustelina, A. sowerbyana, Partulina redfieldi, and Perdicella helena, five species of Hawaiian tree snails across three genera. Mitogenomes ranged in length from 15,187 to 16,793 base pairs, with a base composition of A (36.4-37.4%); T (42.2-42.7%); C (8.8-9.2%); and G (11.3-11.8%). Similar with other pulmonates, these mitogenomes contain 13 protein-coding genes, two ribosomal RNA genes, and 22 transfer RNA genes, with the order conserved among genera. Our study suggests polyphyly in the current arrangement of the subfamily Achatinellinae, part of a spectacular radiation in the Hawaiian Islands.

Diverse habitat use during two life stages of the critically endangered Bahama Oriole (Icterus northropi): community structure, foraging, and social interactions.

Our ability to prevent extinction in declining populations often depends on effective management of habitats that are disturbed through wildfire, logging, agriculture, or development. In these disturbed landscapes, the juxtaposition of multiple habitat types can be especially important to fledglings and young birds, which may leave breeding grounds in human-altered habitat for different habitats nearby that provide increased foraging opportunities, reduced competition, and higher protection from predators. In this study, we evaluated the importance of three habitat types to two life stages of the critically endangered Bahama Oriole (Icterus northropi), a synanthropic songbird endemic to Andros, The Bahamas. First, we determined the avian species composition and relative abundance of I. northropi among three major vegetation types on Andros: Caribbean pine (Pinus caribaea) forest, coppice (broadleaf dry forest), and anthropogenic areas, dominated by nonnative vegetation (farmland and developed land). We then compared the foraging strategies and social interactions of two age classes of adult Bahama Orioles in relation to differential habitat use. Bird surveys late in the Bahama Oriole's breeding season indicated the number of avian species and Bahama Oriole density were highest in coppice. Some bird species occurring in the coppice and pine forest were never observed in agricultural or residential areas, and may be at risk if human disturbance of pine forest and coppice increases, as is occurring at a rapid pace on Andros. During the breeding season, second-year (SY) adult Bahama Orioles foraged in all vegetation types, whereas after-second-year (ASY) adults were observed foraging only in anthropogenic areas, where the species nested largely in introduced coconut palms (Cocos nucifera). Additionally, SY adults foraging in anthropogenic areas were often observed with an ASY adult, suggesting divergent habitat use for younger, unpaired birds. Other aspects of foraging (vegetation features, food-gleaning behavior, and food items) were similar for the two age classes. Older Bahama Orioles exhibited relatively higher rates of social interactions (intraspecific and interspecific pooled) in anthropogenic areas, and won more interaction outcomes compared to younger adults. Our findings concur with those of other studies indicating dry broadleaf forest is vitally important to migrating, wintering, and resident birds, including the critically endangered Bahama Oriole, which appears to depend heavily on this vegetation type during certain life stages.

The complete mitochondrial genome of Achatinella sowerbyana (Gastropoda: Pulmonata: Stylommatophora: Achatinellidae).

In this study, we report the complete mitochondrial genome sequence of Achatinella sowerbyana, an endangered Hawaiian tree snail. The mitogenome is 15,374 bp in length and has a base composition of A (36.4%), T (42.7%), C (9.1%), and G (11.8%). Similar to other pulmonates, it contains 13 protein-coding genes, 2 ribosomal RNA genes, and 22 transfer RNA genes. The gene order is the same as its sister species, A. mustelina. To our knowledge, this is the second mitochondrial genome sequenced within the superfamily Achatinelloidea, and will aid in the examination of the genus Achatinella, part of a spectacular radiation in the Hawaiian Islands.

The complete mitochondrial genome of Achatinella mustelina (Gastropoda: Pulmonata: Stylommatophora).

In this study, we report the complete mitochondrial genome sequence of Achatinella mustelina, an endangered Hawaiian tree snail. The mitogenome is 16 323 bp in length and has a base composition of A (34.7%), T (42.6%), C (12.7%) and G (10.0%). Similar to other Pulmonates, it contains 13 protein-coding genes, 2 rRNA genes and 22 tRNA genes. To our knowledge, this is the first mitochondrial genome sequenced within the Achatinelloidea superfamily, which contains a high number of endangered species. As such, this mitogenome will be useful in conservation genetics studies.

Geographic variation and genetic structure in the Bahama Oriole (Icterus northropi), a critically endangered synanthropic species.

Bird species may exhibit unexpected population structuring over small distances, with gene flow restricted by geographic features such as water or mountains. The Bahama Oriole (Icterus northropi) is a critically endangered, synanthropic island endemic with a declining population of fewer than 300 individuals. It now remains only on Andros Island (The Bahamas), which is riddled with waterways that past studies assumed did not hinder gene flow. We examined 1,858 base pairs of mitochondrial DNA sequenced from four gene regions in 14 birds (roughly 5% of the remaining population) found on the largest land masses of Andros Island (North Andros and Mangrove Cay/South Andros). We sought to discern genetic structuring between the remaining subpopulations and its relationship to current conservation concerns. Four unique haplotypes were identified, with only one shared between the two subpopulations. Nucleotide and haplotype diversity were higher for the North Andros subpopulation than for the Mangrove Cay/South Andros subpopulation. Analysis of molecular variance (AMOVA) yielded a Wright's fixation index (F st) of 0.60 (P Fst = 0.016), with 40.2% of the molecular variation explained by within-population differences and 59.8% by among-population differences. Based on the mitochondrial regions examined in this study, we suggest the extant subpopulations of Bahama Oriole exhibit significant population structuring over short distances, consistent with some other non-migratory tropical songbird species.

Demographic and genetic factors in the recovery or demise of ex situ populations following a severe bottleneck in fifteen species of Hawaiian tree snails.

Wild populations of endangered Hawaiian tree snails have declined precipitously over the last century due to introduced predators and other human impacts. Life history traits, such as very low fecundity (<5 offspring per year across taxa) and maturity at approximately four years of age have made recovery difficult. Conservation efforts such as in situ predator-free enclosures may increase survival to maturity by protecting offspring from predation, but no long-term data existed prior to this study demonstrating the demographic and genetic parameters necessary to maintain populations within those enclosures. We evaluated over 20 years of evidence for the dynamics of survival and extinction in captive ex situ populations of Hawaiian tree snails established from wild-collected individuals. From 1991 to 2006, small numbers of snails (<15) from fifteen species were collected from the wild to initiate captive-reared populations as a hedge against extinction. This small number of founders resulted in a severe bottleneck in each of the captive-reared populations. We identified key demographic parameters that predicted population recovery from this bottleneck. Species with captive populations that produced between two and four offspring per adult per year and had 20-50% of those offspring survive to maturity recovered to numbers above 100 individuals, and maintained viable populations following a decline that occurred between 2009 and 2014. Those populations that had less than two offspring per adult per year and less than 20% survival to maturity did not reach 100 individuals in captivity, and many of these populations died out during the recent decline. We suggest that small reductions in fitness may contribute to extirpation in taxa with inherently low fecundity, by keeping populations below a threshold number essential to long-term recovery. Future ex situ populations should be founded with no less than 15 adults, and maintained in conditions closely approximating the temperature and humidity of source locations to optimize fitness. Permanent translocations of wild populations for conservation purposes will be more likely to succeed with greater than 100 adults, and should be limited to locations with a similar climate to source locations.

Dining local: the microbial diet of a snail that grazes microbial communities is geographically structured.

Achatinella mustelina is a critically endangered tree snail that subsists entirely by grazing microbes from leaf surfaces of native trees. Little is known about the fundamental aspects of these microbe assemblages: not taxonomic composition, how this varies with host plant or location, nor whether snails selectively consume microbes. To address these questions, we collected 102 snail faecal samples as a proxy for diet, and 102 matched-leaf samples from four locations. We used Illumina amplicon sequencing to determine bacterial and fungal community composition. Microbial community structure was significantly distinct between snail faeces and leaf samples, but the same microbes occurred in both. We conclude that snails are not 'picky' eaters at the microbial level, but graze the surface of whatever plant they are on. In a second experiment, the gut was dissected from non-endangered native tree snails in the same family as Achatinella to confirm that faecal samples reflect gut contents. Over 60% of fungal reads were shared between faeces, gut and leaf samples. Overall, location, sample type (faeces or leaf) and host plant identity all significantly explained the community composition and variation among samples. Understanding the microbial ecology of microbes grazed by tree snails enables effective management when conservation requires captive breeding or field relocation.

Population genetics and the effects of a severe bottleneck in an ex situ population of critically endangered Hawaiian tree snails.

As wild populations decline, ex situ propagation provides a potential bank of genetic diversity and a hedge against extinction. These programs are unlikely to succeed if captive populations do not recover from the severe bottleneck imposed when they are founded with a limited number of individuals from remnant populations. In small captive populations allelic richness may be lost due to genetic drift, leading to a decline in fitness. Wild populations of the Hawaiian tree snail Achatinella lila, a hermaphroditic snail with a long life history, have declined precipitously due to introduced predators and other human impacts. A captive population initially thrived after its founding with seven snails, exceeding 600 captive individuals in 2009, but drastically declined in the last five years. Measures of fitness were examined from 2,018 captive snails that died between 1998 and 2012, and compared with genotypic data for six microsatellite loci from a subset of these deceased snails (N = 335), as well as live captive snails (N = 198) and wild snails (N = 92). Surprisingly, the inbreeding coefficient (Fis) declined over time in the captive population, and is now approaching values observed in the 2013 wild population, despite a significant decrease in allelic richness. However, adult annual survival and fecundity significantly declined in the second generation. These measures of fitness were positively correlated with heterozygosity. Snails with higher measures of heterozygosity had more offspring, and third generation offspring with higher measures of heterozygosity were more likely to reach maturity. These results highlight the importance of maintaining genetic diversity in captive populations, particularly those initiated with a small number of individuals from wild remnant populations. Genetic rescue may allow for an increase in genetic diversity in the captive population, as measures of heterozygosity and rarified allelic richness were higher in wild tree snails.