Nest quality predicts the probability of egg loss in the critically endangered 'Alala (Corvus hawaiiensis).

Conservation breeding programs for endangered species face challenges, notably in the development of husbandry techniques, complicated by the impracticality of conducting controlled experiments. To reduce uncertainty regarding what works in conservation breeding programs, it is essential to capture data. In avian breeding programs, the construction of quality nests and appropriate incubation and handling of eggs by the parents are essential prerequisites to the successful production of offspring. Here, we study factors influencing nest-building and parental incubation outcomes in 'alala (Corvus hawaiiensis), which is extinct in the wild and numbers fewer than 130 individuals in human care. Using parent-incubated egg data from 2018 to 2021 (171 clutches, 55 pairs), we evaluated the role of husbandry factors in determining the quality of nests constructed by 'alala and the likelihood of nest quality and other social and environmental factors to predict egg loss (breaking, cannibalization, or ejection of an egg from the nest). More than half of all eggs laid failed to reach the hatch date. Nest quality was the most influential predictor of egg loss, with eggs in higher quality nests more likely to reach the hatch date. Male age also influenced egg loss, with very young (3 years old) and older males (⪞12 years old) experiencing more egg loss. Furthermore, the two facilities and two aviary types also differed in the quality of nests 'alala constructed, indicating that nest-building behavior can be influenced by husbandry practices. These findings reduce uncertainty and provide insights for recommended management interventions to facilitate successful reproduction in human care.

Reduction of genetic diversity in 'Alala (Hawaiian crow; Corvus hawaiiensis) between the late 1800s and the late 1900s.

Genetic and genomic data are increasingly used to aid conservation management of endangered species by providing insights into evolutionary histories, factors associated with extinction risks, and potential for future adaptation. For the 'Alala, or Hawaiian crow (Corvus hawaiiensis), genetic concerns include negative correlations between inbreeding and hatching success. However, it is unclear if low genetic diversity and inbreeding depression are consequences of a historical population bottleneck, or if 'Alala had historically low genetic diversity that predated human influence, perhaps as a result of earlier declines or founding events. In this study, we applied a hybridization-based sequence capture to generate a genome-wide single nucleotide polymorphism (SNP) dataset for comparing historical specimens collected in the 1890s, when 'Alala were more numerous, to samples taken between 1973 and 1998, when 'Alala population densities were near the lowest documented levels in the wild, prior to all individuals being collected for captive rearing. We found low genome-wide diversity in both sample groups, however, the modern sample group (1973 to 1998 cohort) exhibited relatively fewer polymorphic alleles, a lower proportion of polymorphic loci, and lower observed heterozygosity, consistent with a population decline and potential bottleneck effects. These results combined with a current low population size highlight the importance of continued efforts by conservation managers to mitigate inbreeding and maintain founder representation to preserve what genetic diversity remains.

Adapting conservation breeding techniques using a data-driven approach to restore the 'Alala (Hawaiian crow, Corvus hawaiiensis).

For some critically endangered species, conservation breeding is a vital steppingstone toward re-establishing wild populations. The 'Alala (Hawaiian crow, Corvus hawaiiensis), currently extinct in the wild, exists today only in a conservation breeding program, which, for many years, utilized successful hands-on husbandry approaches such as separating and resocializing pairs, providing partially manmade nests, artificially incubating eggs, and puppet rearing nestlings. Yet, a top priority of any conservation breeding program is to retain natural behaviors essential to postrelease survival and reproduction, to achieve successful reintroduction and restoration to the wild. We describe how we are adapting 'Alala husbandry techniques to strengthen pair bonds through full-time socialization, enable pairs to build robust nests, encourage females to incubate eggs to hatch, and provide pairs and their offspring with vital parental rearing experiences. We discuss the use of standardized, data-driven methods to objectively track our progress towards successful parental breeding and to select release candidates based on their likelihood to survive and breed in the wild. The information shared in this report can be applied to other conservation breeding programs, particularly those implementing or transitioning to husbandry techniques geared towards preparing species to thrive in the wild.

Moving from trends to benchmarks by using regression tree analysis to find inbreeding thresholds in a critically endangered bird.

Understanding how inbreeding affects endangered species in conservation breeding programs is essential for their recovery. The Hawaiian Crow ('Alala) (Corvus hawaiiensis) is one of the world's most endangered birds. It went extinct in the wild in 2002, and, until recent release efforts starting in 2016, nearly all of the population remained under human care for conservation breeding. Using pedigree inbreeding coefficients (F), we evaluated the effects of inbreeding on Hawaiian Crow offspring survival and reproductive success. We used regression tree analysis to identify the level of inbreeding (i.e., inbreeding threshold) that explains a substantial decrease in 'Alala offspring survival to recruitment. Similar to a previous study of inbreeding in 'Alala, we found that inbreeding had a negative impact on offspring survival but that parental (vs. artificial) egg incubation improved offspring survival to recruitment. Furthermore, we found that inbreeding did not substantially affect offspring reproductive success, based on the assumption that offspring that survive to adulthood breed with distantly related mates. Our novel application of regression tree analysis showed that offspring with inbreeding levels exceeding F = 0.098 were 69% less likely to survive to recruitment than more outbred offspring, providing a specific threshold value for ongoing population management. Our results emphasize the importance of assessing inbreeding depression across all life history stages, confirm the importance of prioritizing parental over artificial egg incubation in avian conservation breeding programs, and demonstrate the utility of regression tree analysis as a tool for identifying inbreeding thresholds, if present, in any pedigree-managed population.

Moverse de las Tendencias a los Referentes Mediante el Análisis de Árbol de Regresión para Encontrar los Umbrales de Endogamia de una Ave en Peligro Crítico Resumen Es necesario entender cómo afecta la endogamia a las especies en peligro que se encuentran en programas de reproducción para la conservación para la recuperación exitosa de estas especies. El cuervo hawaiano ('Alala) (Corvus hawaiiensis) es una de las especies de ave que se encuentra en mayor peligro de extinción en todo el mundo. Esta especie se extinguió en vida libre en el 2002 y no fue hasta los recientes esfuerzos de liberación que iniciaron en 2016 que casi toda la población permaneció bajo cuidado humano en programas de reproducción para la conservación. Usamos los coeficientes de linaje endogámico (F) para evaluar los efectos de la endogamia sobre el éxito reproductivo y de supervivencia de la descendencia de los cuervos hawaianos. Usamos un análisis de árbol de regresión para identificar el nivel de endogamia (es decir, el umbral de endogamia) que explicara una disminución sustancial en la supervivencia de la descendencia de 'Alala hasta el reclutamiento. Similar a un estudio previo de la endogamia en los 'Alala, descubrimos que la endogamia tuvo un impacto negativo sobre la supervivencia de la descendencia pero que la incubación parental (vs. la artificial) de huevos mejoró la supervivencia de la descendencia hasta el reclutamiento. Además, encontramos que la endogamia no afectó de manera importante al éxito reproductivo de la descendencia, esto basado en la suposición de que los descendientes que sobreviven hasta la edad adulta se reproducen con parejas cuya relación familiar es lejana. Nuestra aplicación novedosa del análisis de árbol de regresión mostró que la descendencia con niveles de endogamia que exceden F = 0.098 tuvieron 69% menos de probabilidad de sobrevivir hasta el reclutamiento que aquella progenie con menor endogamia, lo que proporciona un valor específico de umbral para el manejo continuo de la población. Nuestros resultados resaltan la importancia de la evaluación de la depresión endogámica en todas las etapas de la historia de vida, confirman la importancia de priorizar la incubación parental de huevos sobre la artificial en los programas de conservación de aves y demuestran la utilidad del análisis de árbol de regresión como herramienta para identificar los umbrales de endogamia, cuando presentes, en cualquier población manejada por linaje.

Preliminary observations of tool-processing behaviour in Hawaiian crows Corvus hawaiiensis.

Very few animal species habitually make and use foraging tools. We recently discovered that the Hawaiian crow is a highly skilled, natural tool user. Most captive adults in our experiment spontaneously used sticks to access out-of-reach food from a range of extraction tasks, exhibiting a surprising degree of dexterity. Moreover, many birds modified tools before or during deployment, and some even manufactured tools from raw materials. In this invited addendum article, we describe and discuss these observations in more detail. Our preliminary data, and comparisons with the better-studied New Caledonian crow, suggest that the Hawaiian crow has extensive tool-modification and manufacture abilities. To chart the full extent of the species' natural tool-making repertoire, we have started conducting dedicated experiments where subjects are given access to suitable raw materials for tool manufacture, but not ready-to-use tools.

A High-Quality, Long-Read De Novo Genome Assembly to Aid Conservation of Hawaii's Last Remaining Crow Species.

Abstract: Genome-level data can provide researchers with unprecedented precision to examine the causes and genetic consequences of population declines, which can inform conservation management. Here, we present a high-quality, long-read, de novo genome assembly for one of the world's most endangered bird species, the 'Alala (Corvus hawaiiensis; Hawaiian crow). As the only remaining native crow species in Hawai'i, the 'Alala survived solely in a captive-breeding program from 2002 until 2016, at which point a long-term reintroduction program was initiated. The high-quality genome assembly was generated to lay the foundation for both comparative genomics studies and the development of population-level genomic tools that will aid conservation and recovery efforts. We illustrate how the quality of this assembly places it amongst the very best avian genomes assembled to date, comparable to intensively studied model systems. We describe the genome architecture in terms of repetitive elements and runs of homozygosity, and we show that compared with more outbred species, the 'Alala genome is substantially more homozygous. We also provide annotations for a subset of immunity genes that are likely to be important in conservation management, and we discuss how this genome is currently being used as a roadmap for downstream conservation applications.

Animal Welfare in Conservation Breeding: Applications and Challenges.

Animal welfare and conservation breeding have overlapping and compatible goals that are occasionally divergent. Efforts to improve enclosures, provide enriching experiences, and address behavioral and physical needs further the causes of animal welfare in all zoo settings. However, by mitigating stress, increasing behavioral competence, and enhancing reproduction, health, and survival, conservation breeding programs must also focus on preparing animals for release into the wild. Therefore, conservation breeding facilities must strike a balance of promoting high welfare, while minimizing the effects of captivity to increase population sustainability. As part of the Hawaii Endangered Bird Conservation Program, San Diego Zoo Global operates two captive breeding facilities that house a number of endangered Hawaiian bird species. At our facilities we aim to increase captive animal welfare through husbandry, nutrition, behavior-based enrichment, and integrated veterinary practices. These efforts help foster a captive environment that promotes the development of species-typical behaviors. By using the "Opportunities to Thrive" guiding principles, we outline an outcome-based welfare strategy, and detail some of the related management inputs, such as transitioning to parental rearing, and conducting veterinary exams remotely. Throughout we highlight our evidence-based approach for evaluating our practices, by monitoring welfare and the effectiveness of our inputs. Additionally we focus on some of the unique challenges associated with improving welfare in conservation breeding facilitates and outline concrete future steps for improving and evaluating welfare outcomes that also meet conservation goals.

Discovery of species-wide tool use in the Hawaiian crow.

Only a handful of bird species are known to use foraging tools in the wild. Amongst them, the New Caledonian crow (Corvus moneduloides) stands out with its sophisticated tool-making skills. Despite considerable speculation, the evolutionary origins of this species' remarkable tool behaviour remain largely unknown, not least because no naturally tool-using congeners have yet been identified that would enable informative comparisons. Here we show that another tropical corvid, the 'Alala (C. hawaiiensis; Hawaiian crow), is a highly dexterous tool user. Although the 'Alala became extinct in the wild in the early 2000s, and currently survives only in captivity, at least two lines of evidence suggest that tool use is part of the species' natural behavioural repertoire: juveniles develop functional tool use without training, or social input from adults; and proficient tool use is a species-wide capacity. 'Alala and New Caledonian crows evolved in similar environments on remote tropical islands, yet are only distantly related, suggesting that their technical abilities arose convergently. This supports the idea that avian foraging tool use is facilitated by ecological conditions typical of islands, such as reduced competition for embedded prey and low predation risk. Our discovery creates exciting opportunities for comparative research on multiple tool-using and non-tool-using corvid species. Such work will in turn pave the way for replicated cross-taxonomic comparisons with the primate lineage, enabling valuable insights into the evolutionary origins of tool-using behaviour.

Residue profiles of brodifacoum in coastal marine species following an island rodent eradication.

The second-generation anticoagulant rodenticide brodifacoum is an effective tool for the eradication of invasive rodents from islands and fenced sanctuaries, for biodiversity restoration. However, broadcast application of brodifacoum bait on islands may expose non-target wildlife in coastal marine environments to brodifacoum, with subsequent secondary exposure risk for humans if such marine wildlife is harvested for consumption. We report a case study of monitoring selected marine species following aerial application of brodifacoum bait in August 2011 to eradicate Norway rats (Rattus norvegicus) from Ulva Island, New Zealand. Residual concentrations of brodifacoum were detected in 3 of 10 species of coastal fish or shellfish sampled 43-176d after bait application commenced. Residual brodifacoum concentrations were found in liver, but not muscle tissue, of 2 of 24 samples of blue cod (0.026 and 0.092 µg/g; Parapercis colias) captured live then euthanized for tissue sampling. Residual brodifacoum concentrations were also found in whole-body samples of 4 of 24 mussels (range=0.001-0.022 µg/g, n=4; Mytilus edulis) and 4 of 24 limpets (range=0.001-0.016 µg/g, n=4; Cellana ornata). Measured residue concentrations in all three species were assessed as unlikely to have eventually caused mortality of the sampled individuals. We also conducted a literature review and determined that in eleven previous accounts of residue examination of coastal marine species following aerial applications of brodifacoum bait, including our results from Ulva Island, the overall rate of residue detection was 5.6% for marine invertebrates (11 of 196 samples tested) and 3.1% for fish (2 of 65 samples tested). Furthermore, our results from Ulva Island are the first known detection of brodifacoum residue in fish liver following an aerial application of brodifacoum bait. Although our findings confirm the potential for coastal marine wildlife to be exposed to brodifacoum following island rodent eradications using aerial bait application, the risk of mortality to exposed individual fish or shellfish appears very low. There is also a very low risk of adverse effects on humans that consume fish or shellfish containing residual concentrations in the ranges reported here. Furthermore, any brodifacoum residues that occur in marine wildlife decline to below detectable concentrations over a period of weeks. Thus potential human exposure to brodifacoum through consumption of marine wildlife containing residual brodifacoum could be minimized by defining 'no take' periods for harvest following bait application and regular monitoring to confirm the absence of detectable residues in relevant marine wildlife.