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Carter T. Atkinson

Publications and source records attributed to Carter T. Atkinson.

At least 19 recordsLinked to original sources
USGS2026

Phylogenomic analyses reveal introgression and cryptic speciation in the globally distributed, vector-transmitted pathogen Plasmodium relictum

Establishing species limits is challenging, particularly for pathogens of wildlife. These pathogens can be difficult to sample and culture, and their genome sequencing must often be conducted in the presence of high levels of host DNA. Plasmodium relictum is a mosquito-vectored avian malaria pathogen that is a globally distributed host generalist, comprised of several genetic lineages. We used sequence capture data from 52 P. relictum infections originating from multiple continents to generate a genomic dataset of the pathogen. With this data, we established a robust phylogeny and determined species limits among P. relictum lineages. We generated phylogenomic trees by maximum likelihood and Bayesian methods with multi-species coalescent models and confirmed robustness of the topology by varying the amount of missing data in the analyses. Our results suggest the existence of two cryptic species among the infections we analyzed and provide evidence of genetic introgression between these species. One of the cryptic species, GRW4, devastated the endemic and immunologically naïve avifauna of Hawaii after its introduction to the islands ca. 100 years ago, and so was tested for positive selection in the GRW4 Hawaiian clade. Although we hypothesized it would be released from host selective pressures, we did not find evidence of positive selection in the Hawaiian GRW4 clade, and we discuss possible explanations. Overall, our results underscore the importance of genomic analyses for resolving pathogen species limits and understanding pathogen evolution.

Molecular Phylogenetics and Evolution↗
USGS2024

Environmental and geographical factors influence the occurrence and abundance of the southern house mosquito, Culex quinquefasciatus, in Hawai‘i

Hawaiian honeycreepers, a group of endemic Hawaiian forest birds, are being threatened by avian malaria, a non-native disease that is driving honeycreepers populations to extinction. Avian malaria is caused by the parasite Plasmodium relictum , which is transmitted by the invasive mosquito Culex quinquefasciatus . Environmental and geographical factors play an important role in shaping mosquito-borne disease transmission dynamics through their influence on the distribution and abundance of mosquitoes. We assessed the effects of environmental (temperature, precipitation), geographic (site, elevation, distance to anthropogenic features), and trap type (CDC light trap, CDC gravid trap) factors on mosquito occurrence and abundance. Occurrence was analyzed using classification and regression tree models (CART) and generalized linear models (GLM); abundance (count data) was analyzed using generalized linear mixed models (GLMMs). Models predicted highest mosquito occurrence at mid-elevation sites and between July and November. Occurrence increased with temperature and precipitation up to 580 mm. For abundance, the best model was a zero-inflated negative-binomial model that indicated higher abundance of mosquitoes at mid-elevation sites and peak abundance between August and October. Estimation of occurrence and abundance as well as understanding the factors that influence them are key for mosquito control, which may reduce the risk of forest bird extinction.

Hawaii↗
USGS2024

Linking avian malaria parasitemia estimates from quantitative PCR and microscopy reveals new infection patterns in Hawai'i

Plasmodium parasites infect thousands of species and provide an exceptional system for studying host-pathogen dynamics, especially for multi-host pathogens. However, understanding these interactions requires an accurate assay of infection. Assessing Plasmodium infections using microscopy on blood smears often misses infections with low parasitemias (the fractions of cells infected), and biases in malaria prevalence estimates will differ among hosts that differ in mean parasitemias. We examined Plasmodium relictum infection and parasitemia using both microscopy of blood smears and quantitative polymerase chain reaction (qPCR) on 299 samples from multiple bird species in Hawai'i and fit models to predict parasitemias from qPCR cycle threshold (Ct) values. We used these models to quantify the extent to which microscopy underestimated infection prevalence and to more accurately estimate infection patterns for each species for a large historical study done by microscopy. We found that most qPCR-positive wild-caught birds in Hawaii had low parasitemias (Ct scores ≥35), which were rarely detected by microscopy. The fraction of infections missed by microscopy differed substantially among eight species due to differences in species’ parasitemia levels. Infection prevalence was likely 4–5-fold higher than previous microscopy estimates for three introduced species, including Zosterops japonicus , Hawaii’s most abundant forest bird, which had low average parasitemias. In contrast, prevalence was likely only 1.5–2.3-fold higher than previous estimates for Himatione sanguinea and Chlorodrepanis virens , two native species with high average parasitemias. Our results indicate that relative patterns of infection among species differ substantially from those observed in previous microscopy studies, and that differences depend on variation in parasitemias among species. Although microscopy of blood smears is useful for estimating the frequency of different Plasmodium stages and host attributes, more sensitive quantitative methods, including qPCR, are needed to accurately estimate and compare infection prevalence among host species.

Hawai'i↗
USGS2023

Gene expression reveals immune response strategies of naïve Hawaiian honeycreepers experimentally infected with introduced avian malaria

The unprecedented rise in the number of new and emerging infectious diseases in the last quarter century poses direct threats to human and wildlife health. The introduction to the Hawaiian archipelago of Plasmodium relictum and the mosquito vector that transmits the parasite has led to dramatic losses in endemic Hawaiian forest bird species. Understanding how mechanisms of disease immunity to avian malaria may evolve is critical as climate change facilitates increased disease transmission to high elevation habitats where malaria transmission has historically been low and the majority of the remaining extant Hawaiian forest bird species now reside. Here, we compare the transcriptomic profiles of highly susceptible Hawai‘i ‘amakihi ( Chlorodrepanis virens ) experimentally infected with P. relictum to those of uninfected control birds from a naïve high elevation population. We examined changes in gene expression profiles at different stages of infection to provide an in-depth characterization of the molecular pathways contributing to survival or mortality in these birds. We show that the timing and magnitude of the innate and adaptive immune response differed substantially between individuals that survived and those that succumbed to infection, and likely contributed to the observed variation in survival. These results lay the foundation for developing gene-based conservation strategies for Hawaiian honeycreepers by identifying candidate genes and cellular pathways involved in the pathogen response that correlate with a bird’s ability to recover from malaria infection.

Hawaii↗
USGS2023

Carbon dioxide (CO2) gas and eDNA monitoring as tools for eradicating invasive fish from anchialine pools in Hawai‘i

Invasive fish can profoundly affect communities they invade. In Hawai‘i, invasive fishes have become established in many anchialine pools, threatening the persistence of resident invertebrates, including several endangered species. Tools to eradicate invasive fishes from these pools are lacking. This study tested the efficacy of carbon dioxide (CO2) gas diffused into anchialine pool water as a method to eradicate invasive Mozambique tilapia (Oreochromis mossambicus), guppies (Poecilia reticulata), and western mosquitofish (Gambusia affinis). We first conducted aquarium trials to identify how these fishes were affected by elevated CO2 and the concomitant reduction in pH. We then carried out field trials in pools containing these fish in one pool each at two national historical parks on the Island of Hawai‘i during July 2021–January 2022. We also developed environmental DNA (eDNA) protocols to detect fish that may have survived CO2 treatments. The effect of CO2 on fish behavior varied among species; at pH 5.3 (CO2 = 255 mg/L) for tilapia and 5.0 (CO2 = 488 mg/L) for tilapia, guppies, and mosquitofish, all generally lost their ability to swim, showed slow or no gill movement, and altered their position in the water column. No tilapia survived the trials (n = 4 and 6 individuals at pH 5.3 and 5.0, respectively). In contrast, 41.7% (n = 12) of adult guppies and 66.7% (n = 12) of adult mosquitofish survived treatment at pH 5.0. In the field we were unable to reduce anchialine pool water pH below 5.7. Regardless, we were able to eradicate tilapia from one pool over four sequential treatments. Post-treatment eDNA assessments supported visual surveys, confirming our results. We were not able to eradicate guppies and mosquitofish. Results from this study show that CO2 can be an effective tool for eradicating invasive tilapia from anchialine pools, and post-treatment eDNA assessments can provide managers with a method for evaluating the success of eradication efforts.

Hawaii↗
USGS2022

Environmental monitoring for invasive fungal pathogens of ʽŌhiʽa (Metrosideros polymorpha) on the Island of Hawaiʽi

The invasive rust Austropuccina psidii was detected in the Hawaiian Islands in 2005 and has become widely established throughout the archipelago in both native and introduced species of Myrtaceae. Initial predictions about the impacts of the fungus on native ʽōhiʽa lehua ( Metrosideros polymorpha ), a keystone native tree, have not materialized, but there is ongoing concern that introductions of new genotypes of the fungus could lead to widespread mortality with catastrophic effects on native ecosystems. By contrast, two recently emergent Ascomycete pathogens, Ceratocystis lukuohia ( Ceratocystis wilt of ‘ōhi‘a) and C. huliohia ( Ceratocystis canker of ‘ōhi‘a), collectively known to cause Rapid ʽŌhiʽa Death (ROD), are causing significant mortality in native forests on Hawaiʻi and Kauaʻi Islands, but pathways of spread are still incompletely understood. We used a network of passive environmental samplers for collecting windblown urediniospores of Austropuccina to evaluate the effectiveness of environmental monitoring to detect seasonal and landscape-scale differences in airborne propagules of this rust on Hawai`i Island. The samplers were also used to determine if windborn ambrosia beetle frass or spores of Ceratocystis can spread long distances. We found frequent detections and regional and seasonal differences in numbers of samplers that were positive for urediniospores of Austropuccinia , but little evidence of long-distance airborne dispersal of the ROD-causing fungi. The simple, inexpensive platform for sampling airborne fungal spores that we used may have value as a monitoring tool for detecting spread of airborne fungal pathogens, evaluating habitats for suitability for restoration efforts, and for detecting new pathogen introductions, particularly new Austropuccinia genotypes both in Hawaiʻi and other parts of the world.

Hawaii↗
USGS2021

Transcriptome assembly and differential gene expression of the invasive avian malaria parasite Plasmodium relictum in Hawaiʻi

The malaria parasite Plasmodium relictum (lineage GRW4) was introduced less than a century ago to the native avifauna of Hawaiʻi, where it has since caused major declines of endemic bird populations. One of the native bird species that is frequently infected with GRW4 is the Hawaiʻi ʻamakihi ( Chlorodrepanis virens ). To achieve a better understanding of the transcriptional activities of this virulent parasite, we performed a controlled challenge experiment of 15 ʻamakihi that were infected with GRW4. Blood samples containing malaria parasites were collected at two time points (intermediate and peak infection stages) from host individuals that were either experimentally infected by mosquitoes or inoculated with infected blood. We then used RNA sequencing to assemble a high‐quality blood transcriptome of P. relictum GRW4, allowing us to quantify parasite expression levels inside individual birds. We found few significant differences (one to two transcripts) in GRW4 expression levels between host infection stages and between inoculation methods. However, 36 transcripts showed differential expression levels among all host individuals, indicating a potential presence of host‐specific gene regulation across hosts. To reduce the extinction risk of the remaining native bird species in Hawaiʻi, genetic resources of the local Plasmodium lineage are needed to enable further molecular characterization of this parasite. Our newly built Hawaiian GRW4 transcriptome assembly, together with analyses of the parasite's transcriptional activities inside the blood of Hawaiʻi ʻamakihi, can provide us with important knowledge on how to combat this deadly avian disease in the future.

Hawaii↗
USGS2021

Effectiveness of rapid 'ōhi'a death management strategies at a focal disease outbreak on Hawai'i Island

The ongoing spread of rapid ‘ōhi‘a death (ROD) in the Hawaiian Islands threatens the long-term sustainability of ‘ōhi‘a lehua (Metrosideros polymorpha) forests throughout the state. First identified in the Puna district of Hawai‘i Island in 2014, the disease caused by the novel fungi Ceratocystis lukuohia and Ceratocystis huliohia has now spread island-wide and was recently detected on Kaua‘i, O‘ahu, and Maui. The leading hypothesis for the spread of ROD is through airborne ambrosia beetle frass particles that contain viable Ceratocystis propagules, thus management efforts focus on containing this frass. At the time of this study (2017–2018), the Waipunalei site was the northernmost outbreak of ROD on Hawai‘i Island. The focal nature of the outbreak and accessibility of the location provided the opportunity to monitor the effectiveness of two types of proposed management methods to reduce the airborne spread of potentially infective ambrosia beetle frass: tree felling and insecticide treatments. We placed 23 passive environmental samplers (PES), which monitored for airborne frass and wood particles containing C. lukuohia and C. huliohia in a grid that spanned the outbreak area over 22 weeks. Cross-vane panel traps with 1:1 methanol:ethanol lures were attached to nine of the PES to document wood-boring ambrosia and cerambycid beetle populations during the latter three months of the study. Monitoring with PES began three weeks before management and continued for one month after the last infected trees were felled. Glass microscope slides from the 23 PES were examined for airborne ambrosia beetle frass and wood particles by microscopy. DNA was extracted from the slides and tested by qPCR (quantitative polymerase chain reaction) for C. lukuohia and C. huliohia. We also investigated the correlation of beetle gallery counts with tree height and tested the efficacy of Bifen I/T insecticide (active ingredient: bifenthrin 7.9%) for preventing beetle attacks on the cut surface of ‘ōhi‘a bolts (tree stem sections). Beetle trapping data revealed that the area supports a diverse community of wood-boring beetles, some of which likely attack ‘ōhi‘a and may facilitate the spread of ROD. The number of beetle galleries on felled ‘ōhi‘a trees decreased linearly as tree height increased. We also observed significantly fewer beetle attacks on Bifen I/T treated ‘ōhi‘a bolts than non-treated bolts, but gallery formation nearly ceased in both treated and control bolts by week three. Ceratocystis lukuohia DNA was detected twenty-six times and C. huliohia was detected five times in the PES throughout this study. DNA detections were correlated to frass and wood counts, and the number of felled trees were correlated to wood particle counts but not frass counts. Both the timing and distribution of detections across the sampling grid indicate that tree felling may have reduced airborne detections of Ceratocystis DNA soon after tree felling was completed. A subsequent increase in detections after tree felling ceased may indicate that incomplete removal of infected trees and the appearance of new infections in previously asymptomatic trees could have allowed airborne detections of potentially infectious fungal propagules to once again increase.

Hawaii↗
USGS2020

Introduction to life cycles, taxonomy, distribution and basic research techniques

Avian haemosporidian parasites are a closely related group of apicomplexan parasites with important similarities in their life cycles, development, physiology, and reproduction. Current phylogenies based on mitochondrial and nuclear genes reflect more traditional attempts to classify these organisms based on life history characteristics and morphology, but limited sampling from poorly characterized taxa such as the Garniidae from tropical and subtropical regions continues to limit our understanding of their phylogeny and evolution. Recent advances in molecular diagnostics and the ability to barcode these parasites using mitochondrial cytochrome b sequences have revolutionized the field, but traditional methodology based on microscopy of Giemsa-stained blood smears remains essential for diagnostics and understanding life history characteristics and biodiversity of these organisms. The relative strengths and weaknesses of current methods in wildlife haemosporidian research are discussed. We call for a combination of microscopic, PCR-based, and serological diagnostic methodologies for better estimates of true distribution and other aspects of biology of haemosporidians, particularly in studies on virulence, prevalence, and biodiversity.

Book chapter↗
USGS2020

Use of whole blood samples preserved in DNA lysis buffer for serological detection of avian malaria in Hawaiian forest birds

Recent detections of avian malarial parasites in native and non-native forest birds at Hakalau Forest National Wildlife Refuge and reports of epidemic transmission of the disease in high elevation habitats as well as controversy over accuracy of the PCR (polymerase chain reaction) diagnostic test that was being used led to a request by U.S. Fish and Wildlife Service to see if existing blood samples that were preserved in a DNA lysis buffer could be used for independent confirmation of the findings with antibody based serological methods. The primary objective of this study was to test whether some DNA buffers used for preservation of blood samples cause denaturation and loss of antigenicity of antibody molecules. If the buffer does not destroy antigenicity of these molecules, then the samples can be used in serological assays to provide an independent assessment of the accuracy of PCR tests.

Hawaii↗
USGS2019

Facilitated adaptation for conservation – Can gene editing save Hawaii's endangered birds from climate driven avian malaria?

Avian malaria has played a significant role in causing extinctions, population declines, and limiting the elevational distribution of Hawaiian honeycreepers. Most threatened and endangered honeycreepers only exist in high-elevation forests where the risk of malaria infection is limited. Because Culex mosquito vectors and avian malaria dynamics are strongly influenced by temperature and rainfall, future climate change is predicted to expand malaria infection to high-elevation forests and intensify malaria infection at lower elevations, likely resulting in future extinctions and loss of avian biodiversity in Hawaii. Novel, landscape-level mosquito control strategies are promising, but are logistically challenging and require costly long-term efforts. As an alternative or supplemental strategy, we evaluated the potential of releasing a gene-edited malaria-resistant honeycreeper (Iiwi, Drepanis coccinea) in Hawaiian rainforests; a strategy known as facilitated adaptation. While this approach also has significant technical challenges and costs, it may offer a more permanent solution to increasing malaria threats. If malaria-resistant honeycreepers can be developed, facilitated adaptation may provide a practical strategy for the reestablishment of abundant avian populations in Hawaiian forests. A successful strategy could be the release of malaria-resistant Iiwi in mid-elevation forests where development of a resistant population has the best chance of success. Establishment of a resistant Iiwi population could provide a source for dispersal and development of resistant populations in high-elevation forests and a permanent source of resistant individuals for translocation to other vulnerable areas.

Hawaii↗
USGS2019

Economical environmental sampler designs for detecting airborne spread of fungi responsible for Rapid `Ōhi`a Death

We designed two new samplers for monitoring airborne particulates that rely on either natural wind currents (Passive Environmental Sampler) or a battery-operated fan (Active Environmental Sampler). Both samplers are significantly less expensive than commercial devices such as Rotorod® and Burkard Samplers that are used in the agricultural and health science industries. They are economical enough to be deployed in large numbers across broad landscapes. We evaluated their use for detecting airborne spread of ambrosia beetle frass that may contain infective spores of the fungi (Ceratocystis lukuohia and C. huliohia) that are responsible for Rapid `Ōhi`a Death (ROD), a newly documented pathosystem on Hawai`i Island. We compared performance of the new samplers to Rotorod® Model 20 Samplers by releasing synthetic polyethylene spheres (12–160 µm in diameter) and also Xyleborus spp. frass known to contain C. lukuohia and C. huliohia propagules under controlled laboratory and field conditions. Overall, the Active Environmental Sampler proved to be 3–4 times more effective in capturing polyethylene spheres and 2–3 times more effective in capturing frass than either the Passive or Rotorod® Samplers. Significant differences between the Passive and Rotorod® Samplers were not detected. For the frass release experiment, C. lukuohia DNA was detected once by qPCR in an Active Environmental Sampler and C. huliohia DNA was detected during two different trials, once with an Active Environmental Sampler and once with a Passive Environmental Sampler. No detections were made with Rotorod® Samplers. Both Active and Passive Samplers were used in the field for detection of airborne dispersal of C. lukuohia and C. huliohia at Orchidlands Estates in the Puna District of Hawai`i Island. We found that airborne dispersal of potentially infective beetle frass was uncommon over short distances with qPCR detections in up to 10% of weekly sampler collections.

Hawai`i Cooperative Studies Unit Technical Report↗
USGS2018

The epidemiology of avian pox and interaction with avian malaria in Hawaiian forest birds

Despite the purported role of avian pox ( Avipoxvirus spp.) in the decline of endemic Hawaiian birds, few studies have been conducted on the dynamics of this disease, its impact on free‐living avian populations, or its interactions with avian malaria ( Plasmodium relictum ). We conducted four longitudinal studies of 3–7 yr in length and used generalized linear models to evaluate cross‐sectional prevalence of active pox infection and individuals with healed deformities that had recovered from pox. Our goal was to understand how species, season, elevation, malaria infection, and other biological characteristics influenced pox infection in ʻApapane, Hawaiʻi ʻAmakihi, ʻIʻiwi, and Japanese White‐eye across low‐, mid‐, and high‐elevation forests on the island of Hawaiʻi. We also used multi‐state capture‐recapture (longitudinal) models to estimate pox infection rates, recovery rates, and potential pox‐related mortality. Pox infection rates were typically highest in low‐elevation forests, followed by mid‐elevation forests, and lowest in high‐elevation forests. We also found seasonal changes in pox prevalence throughout the annual cycle; typically increasing from spring through summer, peaking in fall, and declining in winter. These seasonal changes occurred in low‐ and mid‐elevation forests, but not in high elevations where pox infection was low. Seasonal and elevation patterns of pox infection are like those for avian malaria, strongly implicating mosquito vectors, rather than other biting arthropods or contact transmission, as the primary source of transmitting both diseases. Most native Hawaiian birds recovered from pox infection within 6 months; frequently without permanent lesions. Contrary to our expectations, we found no direct evidence that pox is a substantial mortality factor in any of the three native bird species we studied. Birds with chronic malaria infection were more likely to have both active pox infection and healed pox lesions suggesting a synergistic interaction that may influence the evolution of pox virulence. Because pox infection can be assessed visually, and birds have a high recovery rate, this disease may be a sensitive indicator of the seasonal and annual risk of transmission of malaria in Hawaiʻi.

Ecological Monographs↗
USGS2018

Characterization of Plasmodium relictum, a cosmopolitan agent of avian malaria

Background Microscopic research has shown that Plasmodium relictum is the most common agent of avian malaria. Recent molecular studies confirmed this conclusion and identified several mtDNA lineages, suggesting the existence of significant intra-species genetic variation or cryptic speciation. Most identified lineages have a broad range of hosts and geographical distribution. Here, a rare new lineage of P. relictum was reported and information about biological characters of different lineages of this pathogen was reviewed, suggesting issues for future research. Methods The new lineage pPHCOL01 was detected in Common chiffchaff Phylloscopus collybita, and the parasite was passaged in domestic canaries Serinus canaria . Organs of infected birds were examined using histology and chromogenic in situ hybridization methods. Culex quinquefasciatus mosquitoes, Zebra finch Taeniopygia guttata, Budgerigar Melopsittacus undulatus and European goldfinch Carduelis carduelis were exposed experimentally. Both Bayesian and Maximum Likelihood analyses identified the same phylogenetic relationships among different, closely-related lineages pSGS1, pGRW4, pGRW11, pLZFUS01, pPHCOL01 of P. relictum. Morphology of their blood stages was compared using fixed and stained blood smears, and biological properties of these parasites were reviewed. Results Common canary and European goldfinch were susceptible to the parasite pPHCOL01, and had markedly variable individual prepatent periods and light transient parasitaemia. Exo-erythrocytic and sporogonic stages were not seen. The Zebra finch and Budgerigar were resistant. Neither blood stages nor vector stages of all examined P. relictum lineages can be distinguished morphologically. Conclusion Within the huge spectrum of vertebrate hosts, mosquito vectors, and ecological conditions, different lineages of P. relictum exhibit indistinguishable, markedly variable morphological forms. Parasites of same lineages often develop differently in different bird species. Even more, the variation of biological properties (parasitaemia dynamics, blood pathology, prepatent period) in different isolates of the same lineage might be greater than the variation in different lineages during development in the same species of birds, indicating negligible taxonomic value of such features. Available lineage information is excellent for parasite diagnostics, but is limited in predictions about relationships in certain host-parasite associations. A combination of experiments, field observations, microscopic and molecular diagnostics is essential for understanding the role of different P. relictum lineages in bird health.

Malaria Journal↗
USGS2018

Population differences in susceptibility to Plasmodium relictum in zebra finches Taeniopygia guttata

Domesticated Australian and Timor zebra finches (Taeniopygia guttata castanotis, and T. guttata guttata, respectively) were inoculated with canary (Serinus canaria) blood containing a Hawaiian isolate of Plasmodium relictum (lineage GRW04), a hemoparasite that causes avian malaria. In two experimental trials, Timor, but not Australian zebra finches developed parasitemia that was detected by microscopic examination of blood smears. In the second trial, in which molecular detection methods were used, a single Australian zebra finch and 5 of 6 challenged Timor birds were positive for the parasite. Additionally, P. relictum DNA was detected in multiple blood samples obtained from Timor birds over the 28 days following challenge. Timor zebra finches may provide a useful, easily maintained, laboratory model for the study of arbovirus/Plasmodium interactions in passerines, but are still inferior to canaries, the traditionally used model of avian malaria infection, in terms of supporting high parasitemia infections.

Avian Diseases↗
USGS2017

Mitigating future avian malaria threats to Hawaiian forest birds from climate change

Avian malaria, transmitted by Culex quinquefasciatus mosquitoes in the Hawaiian Islands, has been a primary contributor to population range limitations, declines, and extinctions for many endemic Hawaiian honeycreepers. Avian malaria is strongly influenced by climate; therefore, predicted future changes are expected to expand transmission into higher elevations and intensify and lengthen existing transmission periods at lower elevations, leading to further population declines and potential extinction of highly susceptible honeycreepers in mid- and high-elevation forests. Based on future climate changes and resulting malaria risk, we evaluated the viability of alternative conservation strategies to preserve endemic Hawaiian birds at mid and high elevations through the 21 st century. We linked an epidemiological model with three alternative climatic projections from the Coupled Model Intercomparison Project to predict future malaria risk and bird population dynamics for the coming century. Based on climate change predictions, proposed strategies included mosquito population suppression using modified males, release of genetically modified refractory mosquitoes, competition from other introduced mosquitoes that are not competent vectors, evolved malaria-tolerance in native honeycreepers, feral pig control to reduce mosquito larval habitats, and predator control to improve bird demographics. Transmission rates of malaria are predicted to be higher than currently observed and are likely to have larger impacts in high-elevation forests where current low rates of transmission create a refuge for highly-susceptible birds. As a result, several current and proposed conservation strategies will be insufficient to maintain existing forest bird populations. We concluded that mitigating malaria transmission at high elevations should be a primary conservation goal. Conservation strategies that maintain highly susceptible species like Iiwi ( Drepanis coccinea ) will likely benefit other threatened and endangered Hawai’i species, especially in high-elevation forests. Our results showed that mosquito control strategies offer potential long-term benefits to high elevation Hawaiian honeycreepers. However, combined strategies will likely be needed to preserve endemic birds at mid elevations. Given the delay required to research, develop, evaluate, and improve several of these currently untested conservation strategies we suggest that planning should begin expeditiously.

PLoS ONE↗
USGS2017

A rapid diagnostic test and mobile "lab in a suitcase" platform for detecting Ceratocystis spp. responsible for Rapid ‘Ōhi‘a Death

We describe a field compatible molecular diagnostic test for two new species of Ceratocystis that infect `ōhi`a (Metrosideros polymorpha) and cause the disease commonly known as Rapid `Ōhi`a Death. The diagnostic is based on amplification of a DNA locus within the internal transcribed spacer region that separates fungal 5.8S ribosomal genes. The assay uses forward and reverse primers, recombinase polymerase, and a fluorescent probe that allows isothermal (40oC) amplification and simultaneous quantification of a 115 base pair product with a battery operated fluorometer. DNA extractions are field compatible and can be done by heating wood drill shavings to 100oC in Instagene® solution containing Chelex® resin to bind potential amplification inhibitors. The initial heat treatment is followed by a short bead beating step with steel ball bearings and zirconium beads to release DNA. DNA is subsequently purified with a magnetic bead based extraction method that does not require silica columns or centrifugation. The assay is designed around a portable “lab-in-a-suitcase” platform that includes a portable fluorometer, miniature centrifuge, and heat block that operate off either 120V AC power sources or a 12 volt battery with a portable inverter, a magnetic rack designed for 1.5 ml tubes and magnetic bead DNA purification, pipettes and consumable reagents and tubes. The entire assay from DNA extraction to results can be performed in less than 90 minutes on up to six independent samples plus a positive and negative control. Sensitivity based on suspensions of Ceratocystis endoconidia (spores) that were added to wood shavings and processed under field conditions by Instagene® magnetic bead DNA extraction was up to 163 spores/mg wood for Species A and 55 spores/mg wood for Species B in 95% of replicates as determined by probit analysis. Sensitivity increased 5–10 fold to 19 spores/mg wood for Species A and 9 spores/mg wood for Species B when extractions were performed with a commercial, silica column based DNA purification kit. The test did not cross react with other common fungi that have been isolated from `ōhi`a.

Technical Report↗