Wild gut microbiomes reveal individuals, species, and location as drivers of variation in two critically endangered Hawaiian honeycreepers.

BACKGROUND: The gut microbiome of animals is an important component that has strong influence on the health, fitness, and behavior of its host. Most research in the microbiome field has focused on human populations and commercially important species. However, researchers are now considering the link between endangered species conservation and the microbiome. In Hawai'i, several threats (e.g., avian malaria and habitat loss) have caused widespread population declines of Hawaiian honeycreepers (subfamily: Carduelinae). These threats can have a significant effect on the avian gut microbiome and may even lead to disruption of microbial function. However, the gut microbiome of honeycreeper in the wild has yet to be explored. METHODS: We collected 13 and 42 fecal samples, respectively, from two critically endangered honeycreeper species, the 'akikiki (Oreomystis bairdi) and the 'akeke'e (Loxops caeruleirostris). The 16S rRNA gene was sequenced and processed though a MOTHUR-based bioinformatics pipeline. Bacterial ASVs were identified using the DADA2 program and bacterial community analyses, including alpha and beta diversity measures, were conducted using R packages Phyloseq and vegan. RESULTS: A total of 8,958 bacterial ASVs were identified from the fecal samples. Intraspecific differences in the gut microbiome among individual birds explained most of the variation present in the dataset, however differences between species did exist. Both species had distinct microbiomes with minimal overlap in beta diversity. 'Akikiki had a more diverse microbiome compared to 'akeke'e. Additionally, small but stastically significant differences in beta diversity also exist between sampling location and sexes in 'akikiki. CONCLUSION: 'Akikiki and 'akeke'e are currently the focus of captive breeding efforts and plans to translocate the two species to other islands are underway. This baseline knowledge will help inform management decisions for these honeycreeper species in their native habitats, on other islands, and in captivity.

Loss of cultural song diversity and the convergence of songs in a declining Hawaiian forest bird community.

The effects of population decline on culturally transmitted behaviours in animals have rarely been described, but may have major implications to population viability. Learned vocal signals in birds are of critical importance to behaviours associated with reproduction, intrasexual interactions and group cohesion, and the complexity of vocal signals such as song can serve as an honest signal of an individual's quality as well as the viability of a population. In this study, we examined how rapid population declines recently experienced by Hawaiian honeycreepers on the island of Kaua'i (USA) may have influenced the diversity, complexity and similarity of learned honeycreeper songs. We analysed the acoustic characteristics of songs recorded during three time periods over a 40-year time frame for three species of declining Kaua'i honeycreepers. We detected a loss of song complexity and diversity over the 40-year time period that paralleled dramatic population declines. Concurrent with the loss of complexity, we also found that the acoustic characteristics of the three honeycreepers' songs became more similar to one another. To our knowledge, this is the first documentation of convergence of acoustic characteristics among rapidly declining species. The reduction in song complexity and diversity and convergence of songs not only signals a loss of culturally transmitted behaviours in these endemic Hawaiian honeycreepers, but also potential challenges to the recovery of these rapidly declining species. Moreover, the present study highlights that there is a 'hidden' cost to declining populations beyond just the loss of individuals that is not often considered, the loss of culturally transmitted social behaviours.

Using population viability analysis to evaluate management activities for an endangered Hawaiian endemic, the Puaiohi (Myadestes palmeri).

Evolution in the Hawaiian Islands has produced a unique avian assemblage. Unfortunately, many of these bird species are highly endangered or extinct. Despite numerous and increasing threats and great effort aimed at saving endemic birds, we lack basic science necessary for understanding many species of concern. One such species is the critically endangered Puaiohi (Myadestes palmeri), a rare songbird endemic to the island of Kaua'i and the only remaining native thrush on the island. At present, the Puaiohi's breeding population is estimated to be ~500 birds restricted to the Alaka'i Wilderness Preserve. We collected demographic data from 2007-2012 and supplemented it with published sources. Using Vortex, we developed stochastic population models to represent Puaiohi population dynamics under current and potential management scenarios to determine management's potential efficacy in aiding species recovery. Management scenarios modeled included rat control, habitat improvement, general survival facilitation, and provision of nest boxes. The model indicated a decline in abundance with a growth rate (r) of -0.267 under baseline conditions. Female and juvenile survival appeared to be the most influential parameters related to population growth and persistence, so management should focus on increasing female and juvenile Puaiohi survival. Rat control, even at more conservative levels, appeared to be the most effective method of increasing Puaiohi abundance. Our results indicate that practical, attainable management activities can increase Puaiohi and bring the species back from the brink of extinction. Such findings provide an example for other endangered species conservation efforts.

Potential disruption of seed dispersal in the absence of a native Kauai thrush.

Hawaii has experienced a catastrophic decline in frugivorous native birds coupled with the introduction of non-native species. Puaiohi (Myadestes palmeri), a critically endangered thrush, is the sole extant native songbird capable of dispersing fleshy fruited plants in the rainforest of Kauai island, Hawaii. As this species has declined to occupy a small proportion of its original range, a suite of largely omnivorous non-native birds have been introduced to this region, including the common and widespread Japanese White-eye (Zosterops japonicus). This reshuffling of the bird community could have long-term implications for plant community composition if introduced birds incompletely replace the ecological role of native species. The objective of this study was to evaluate the potential consequences of the local extirpation of Puaiohi for seed dispersal. Specifically, we compared the diet of Puaiohi and Japanese White-eye, vegetation characteristics, and seed rain at sites with and without Puaiohi in the Na Pali-Kona Forest Reserve on the island of Kauai. We found high overlap in the composition of seeds consumed by the two bird species, but differences in the characteristics of seeds consumed; Japanese White-eye appeared more likely to consume smaller seeded species compared with Puaiohi. Sites with Puaiohi received substantially higher seed rain during the study period, despite no significant differences in overall fruit abundance. Our results suggest that non-native birds are unlikely to completely replace the seed dispersal services provided by Puaiohi. If Puaohi continue to be rare and range restricted, we predict a shift in plant community composition through an increase in non-native and small-seeded plants, and possible dispersal failure of other native species. Our findings lend further support to efforts to conserve Puaiohi across its current and former range, and to consider introductions to other suitable areas to ensure the persistence not only of the species and but also its functional role in Hawaii's montane ecosystems.

Collapsing avian community on a Hawaiian island.

The viability of many species has been jeopardized by numerous negative factors over the centuries, but climate change is predicted to accelerate and increase the pressure of many of these threats, leading to extinctions. The Hawaiian honeycreepers, famous for their spectacular adaptive radiation, are predicted to experience negative responses to climate change, given their susceptibility to introduced disease, the strong linkage of disease distribution to climatic conditions, and their current distribution. We document the rapid collapse of the native avifauna on the island of Kaua'i that corresponds to changes in climate and disease prevalence. Although multiple factors may be pressuring the community, we suggest that a tipping point has been crossed in which temperatures in forest habitats at high elevations have reached a threshold that facilitates the development of avian malaria and its vector throughout these species' ranges. Continued incursion of invasive weeds and non-native avian competitors may be facilitated by climate change and could also contribute to declines. If current rates of decline continue, we predict multiple extinctions in the coming decades. Kaua'i represents an early warning for the forest bird communities on the Maui and Hawai'i islands, as well as other species around the world that are trapped within a climatic space that is rapidly disappearing.

Local prevalence and transmission of avian malaria in the Alakai Plateau of Kauai, Hawaii, U.S.A.

Avian malaria is among the most important threats to native Hawaiian forest birds. It is caused by the parasite Plasmodium relictum and is transmitted by the introduced mosquito vector Culex quinquefasciatus. Temperature increases and precipitation declines due to climate change over the last decade may be responsible for the observed recent expansion in the range and prevalence of avian malaria on the Alakai Plateau, Kauai Island. To examine the hypothesis that conditions are now favorable for transmission of malaria on the Plateau, mosquitoes were sampled with CO2 and Reiter oviposition traps at three sites (Kawaikoi, Halepa'akai, and Koke'e) on several occasions between October, 2013 and April, 2014. P. relictum infection was assessed by PCR or dissection under a microscope. We also surveyed mosquito larvae along Halepa'akai and Kawaikoi streams. We observed that Cx. quinquefasciatus is well established on the Alakai Plateau, as mosquitoes were caught on all field trips, except in April at Halepa'akai, and larvae were found throughout the year. We observed differences in adult abundance among sites and microhabitats (stream vs ridge lines).

Changing climate and the altitudinal range of avian malaria in the Hawaiian Islands - an ongoing conservation crisis on the island of Kaua'i.

Transmission of avian malaria in the Hawaiian Islands varies across altitudinal gradients and is greatest at elevations below 1500 m where both temperature and moisture are favorable for the sole mosquito vector, Culex quinquefasciatus, and extrinsic sporogonic development of the parasite, Plasmodium relictum. Potential consequences of global warming on this system have been recognized for over a decade with concerns that increases in mean temperatures could lead to expansion of malaria into habitats where cool temperatures currently limit transmission to highly susceptible endemic forest birds. Recent declines in two endangered species on the island of Kaua'i, the 'Akikiki (Oreomystis bairdi) and 'Akeke'e (Loxops caeruleirostris), and retreat of more common native honeycreepers to the last remaining high elevation habitat on the Alaka'i Plateau suggest that predicted changes in disease transmission may be occurring. We compared prevalence of malarial infections in forest birds that were sampled at three locations on the Plateau during 1994-1997 and again during 2007-2013, and also evaluated changes in the occurrence of mosquito larvae in available aquatic habitats during the same time periods. Prevalence of infection increased significantly at the lower (1100 m, 10.3% to 28.2%), middle (1250 m, 8.4% to 12.2%), and upper ends of the Plateau (1350 m, 2.0% to 19.3%). A concurrent increase in detections of Culex larvae in aquatic habitats associated with stream margins indicates that populations of the vector are also increasing. These increases are at least in part due to local transmission because overall prevalence in Kaua'i 'Elepaio (Chasiempis sclateri), a sedentary native species, has increased from 17.2% to 27.0%. Increasing mean air temperatures, declining precipitation, and changes in streamflow that have taken place over the past 20 years are creating environmental conditions throughout major portions of the Alaka'i Plateau that support increased transmission of avian malaria.