Locating Nesting Sites for Critically Endangered Galápagos Pink Land Iguanas (Conolophus marthae).

Invasive alien species control is recognized worldwide as a priority action to preserve global biodiversity. However, a lack of general life history knowledge for threatened species can impede the effectiveness of conservation actions. Galápagos pink land iguanas (Conolophus marthae) are endemic to Wolf Volcano, Galápagos, Ecuador. These iguanas are threatened by invasive alien species, particularly feral cats, that may affect their small population size. To guarantee the long-term survival of C. marthae, the Galápagos National Park Directorate is considering, along with an ongoing campaign of feral cat control, the implementation of a head-start program. However, the success of this management strategy necessarily relies on the identification of pink iguana nesting grounds, which were still unknown at the onset of this study. We modeled the movement patterns of male and female iguanas during the reproductive season, using location data collected from custom-made remote tracking devices installed on adult pink iguanas in April 2021. We first calculated for each individual the vector of distances from its starting location, which was defined as net displacement. We then used net displacement as the response variable in a generalized additive mixed model with day of the year as the predictor. Based on the hypothesis that males and females may behaviorally differ after mating, we looked for female-specific migratory behavior suggesting females were moving toward nesting areas. The results obtained confirmed our hypothesis, as females exhibited a distinct migratory behavior, reaching a small plateau area inside of Wolf Volcano's caldera and ca. 400 m below the volcano's northern rim. Moreover, once inside the caldera, females displayed a more aggregated distribution pattern. The movement data obtained allowed Galápagos National Park rangers to locate individual pink iguana nests and subsequently to sight and collect the first observed hatchlings of the species. This work constitutes a necessary baseline to perform dedicated studies of pink iguana nests and emerging hatchling iguanas, which is an essential step toward the development of an effective head-start program.

Habitat restoration to conserve the Little Vermilion Flycatcher on Santa Cruz Island, Galapagos.

The endemic Little Vermilion Flycatcher (LVF), Pyrocephalus nanus, has suffered a drastic decline on Santa Cruz Island, where it was common 30 years ago. Currently, less than 40 individuals remain in the last remnants of natural humid forest in the Galapagos National Park on this island. This small population has low reproductive success, which is contributing to its decline in Santa Cruz. Previous studies have identified Avian Vampire Fly, Philornis downsi, parasitism, changes in food sources, and habitat alteration as threats to this species. In Santa Cruz, invasive plants may strongly affect the reproductive success of the LVF because they limit accessibility to prey near the ground, the preferred foraging niche of these birds. Since 2019, we restored the vegetation in seven plots of one hectare each by removing invasive blackberry plants and other introduced plant species. In all nests that reached late incubation, we also reduced the number of Avian Vampire Fly larvae. In this study, we compared foraging and perch height, pair formation, incubation time and reproductive success between managed and unmanaged areas. As predicted, we found significantly lower foraging height and perch height in 2021 in managed areas compared to unmanaged areas. In 2020, daily failure rate (DFR) of nests in the egg stage did not differ between management types; however, in 2021, DFR in the egg stage was significantly lower in managed areas than in unmanaged areas. The DFR during the nestling stage was similar between managed and unmanaged areas in 2020, but in 2021, only nests in managed areas reached the nestling stage. Females brooded significantly more during the incubation phase in managed areas. Additionally, we found significantly higher reproductive success in managed areas compared to unmanaged areas in 2021, but not in 2020. Habitat restoration is a long-term process and these findings suggest that habitat management positively affects this small population in the long term.

Using modified trapping regimes to understand the behavioral and spatial ecology of Philornis downsi (Diptera: Muscidae).

The avian vampire fly Philornis downsi (Dodge & Aitken) (Diptera: Muscidae) is native to continental South America and the Caribbean, but invasive in the Galapagos Archipelago. The larvae of P. downsi feed on the blood and tissues of the nestlings of 75% of the small land bird species that are endemic or native to Galapagos, causing high in-nest mortality and severe population declines in some species. Efficient trapping techniques are vital to safeguarding these birds in the short term as well as for monitoring fly populations, but basic information about the ecology of the fly is still needed to help develop a species-appropriate trapping method. In this study, we used a novel trapping regime with a vertical distribution to make inferences about P. downsi's behavioral and spatial ecology and to optimize trap catch. Our results showed that male and female P. downsi were trapped in greater numbers below the canopy (3.1-7.5 m), lower down than other commonly caught insect species (5.1-11.5 m). Notably, the effect of trap height remained consistent across seasons and different weather conditions. These findings suggest that P. downsi tend to move at heights where their hosts nest (at or below the canopy) and do not spend time above the canopy. This also makes it unlikely that strategies such as hill-topping or aerial swarming are being used to locate mates. As such, trapping and control efforts should be focused below the canopy in forests with similar canopy heights to effectively capture P. downsi and reduce bycatch of other insects.

Novel herpesvirus in the critically endangered Galapagos pink land iguana.

Virus surveillance in wildlife is important to understanding ecosystem health, taxonomy, and evolution. Nevertheless, viruses in reptiles, and specifically in squamates, continue to be understudied. Herein, we conducted a health assessment on the critically endangered Galapagos pink land iguana (Conolophus marthae) and the vulnerable Galapagos land iguana (Conolophus subcristatus). We collected oral and/or cloacal swabs from 47 clinically healthy iguanas and tested for adenovirus (cloacal swabs, n = 47) and herpesvirus (oral swabs, n = 45) using broad-spectrum PCRs. Two out of 38 (5.3 %) Galapagos pink land iguanas tested positive for herpesvirus, while no herpesvirus was detected in all Galapagos land iguanas (n = 7). Both herpesviral sequences were identical between them and divergent (61.9 % amino acid identity) when compared to the closest herpesvirus sequences available in GenBank/EMBL/DDBJ. The genetic distance between this and other herpesviruses is consistent with its classification as a novel virus species. All iguanas were negative for adenovirus. This is the first description of a herpesvirus in iguanas of the Galapagos islands, and the first report of a potential pathogen for the iconic Galapagos pink land iguana. Further research is needed to understand the implications of this virus in the conservation and management of one of the most endangered iguana species in the world.

Enhancing restoration success of rare plants in an arid-tropical climate through water-saving technologies: a case study of Scalesia affinis ssp. brachyloba in the Galapagos Islands.

Arid tropical archipelagos, such as the Galapagos Islands, host a high concentration of endemic plant species, many of which require restoration intervention to recover from past environmental degradation. Water-saving technologies (WSTs) have potential for hastening restoration by providing plants with additional water during the early stages of growth. However, it remains unclear whether such technologies provide an advantage for plant species of arid-tropical regions. This study examined the effect of the water-saving technology Groasis Waterboxx® (Groasis) on the rare endemic plant species Scalesia affinis ssp. brachyloba during early stages of restoration. Survival was monitored for 374 individuals planted across six sites on Santa Cruz Island, Galapagos (326 with technology and 48 as controls). Kaplan-Meier survival analysis showed that the use of Groasis reduced mortality during the first two years of the seedling survival. A mixed-effect logistic regression that modelled plant survival as a function of total precipitation, maximum temperature, and WST treatment (Groasis and no-technology control) found that despite low overall survival rates, plants grown with Groasis exhibited a three-fold higher predicted survival by the end of the 3.7 year duration of the study. Finally, through a resampling method, we demonstrate that the effect of the WST treatment is not dependent on the unbalanced design typical of a restoration project framework. We conclude that water-saving technologies such as the Groasis Waterboxx® can enhance survival of rare plant species such as S. affinis ssp. brachyloba in restoration programs in arid-tropical regions.

Invasive blackberry outcompetes the endemic Galapagos tree daisy Scalesia pedunculata.

Island florae are under threat from habitat loss and competition with introduced species worldwide. In the Galapagos Islands, the endemic tree daisy Scalesia pedunculata (Asteraceae) is the dominant tree in the cloud forest of Santa Cruz Island but suffers from competition with the invasive blackberry Rubus niveus. At the site Los Gemelos, a S. pedunculata population was monitored from 2014 to 2021 following mechanical and chemical removal of R. niveus from 17 plots and compared with 17 additional plots where R. niveus remained. The aim of this study was to evaluate the impacts of the R. niveus invasion on S. pedunculata by characterizing the effects of R. niveus removal. Parameters measured in S. pedunculata were diameter at breast height (DBH, from which annual growth rates were determined), total height, survival of individual plants, and recruitment. In the presence of R. niveus, S. pedunculata trees had smaller DBH stems and shorter asymptotic maximum heights, growth rates declined for thin trees, the mortality of larger trees was elevated, and S. pedunculata recruitment was absent. R. niveus removal resulted in DBH-ratios of S. pedunculata more frequently meeting our threshold for fast growth (1.2), trees growing significantly thicker and taller, annual mortality being lower (12.5% vs. 16.2% per year), and recruitment being successful. In the presence of R. niveus, lower survival, growth, and absent recruitment suggested that S. pedunculata could reach quasi-extinction in ~20 years. Swift and decisive management action is needed to prevent the Scalesia forest on Santa Cruz Island from disappearing in less than two decades.

Nitrogen and carbon stable isotope analysis sheds light on trophic competition between two syntopic land iguana species from Galápagos.

Coexistence between closely related species can lead to intense competition for resources. Stable isotope analysis (SIA) is a reliable tool to estimate the extent of species competition. We employed SIA to evaluate niche partitioning among two syntopic species of Galápagos land iguanas: Conolophus subcristatus and C. marthae. Samples were collected on Wolf Volcano, Isabela Island, where C. marthae is endemic and syntopic with C. subcristatus. We determined δ13C and δ15N ratios and described the isotopic niche of each species using corrected standard ellipse area (SEAc). We tested for differentiation between the isotopic niches, while controlling for sex, body size, spatial location of samples and mean annual primary productivity at capture points, using bivariate linear models. Despite the extensive overlap of the isotopic niches, we found species and sex to be a significant, interacting predictor of a sample's location in the δ13C, δ15N space, indicating the existence of niche partitioning mechanisms acting between species and sexes. We also found that body size and productivity at the capture points, compounded with yet undetermined spatial effects, explain ca. 75% of the differences observed between species and sexes, providing evidence for differential microhabitat and food-items usage. Our study provides essential baselines for evaluating conservation actions for C. marthae, such as the potential translocation to a sanctuary area free of competition from C. subcristatus.

25-Hydroxyvitamin D Plasma Levels in Natural Populations of Pigmented and Partially Pigmented Land Iguanas from Galápagos (Conolophus spp.).

We report the first data on 25-hydroxyvitamin D plasma levels in natural populations of three species of land iguana endemic to the Galápagos Islands (Conolophus marthae, C. subcristatus, and C. pallidus). The pigment is present throughout the whole body in the skin of C. subcristatus and C. pallidus. On the contrary, pigment is not present in the skin of an extended part of the body in C. marthae. The only existing population of C. marthae is syntopic with a population of C. subcristatus, and the two species are closely related. These circumstances would suggest that, under the assumption that the species show a similar basking behavior and in the absence of compensatory mechanisms, lighter pigmentation should favor higher vitamin D levels. Thus, C. marthae, compared with C. subcristatus in Wolf Volcano, could show higher levels of 25(OH)D plasma levels, or equal, if compensatory mechanisms exist. The three species showed levels in the range of average values for healthy iguanas. However, contrary to the expectation, C. marthae consistently exhibited the lowest 25(OH)D plasma levels. We discuss possible factors affecting vitamin concentration and hypothesize that C. marthae may use the habitat to limit exposure to the high UVB irradiation at Wolf Volcano.

The Galapagos giant tortoise Chelonoidis phantasticus is not extinct.

The status of the Fernandina Island Galapagos giant tortoise (Chelonoidis phantasticus) has been a mystery, with the species known from a single specimen collected in 1906. The discovery in 2019 of a female tortoise living on the island provided the opportunity to determine if the species lives on. By sequencing the genomes of both individuals and comparing them to all living species of Galapagos giant tortoises, here we show that the two known Fernandina tortoises are from the same lineage and distinct from all others. The whole genome phylogeny groups the Fernandina individuals within a monophyletic group containing all species with a saddleback carapace morphology and one semi-saddleback species. This grouping of the saddleback species is contrary to mitochondrial DNA phylogenies, which place the saddleback species across several clades. These results imply the continued existence of lineage long considered extinct, with a current known population size of a single individual.

Conservation of a flagship species: Health assessment of the pink land iguana, Conolophus marthae.

The pink land iguana, Conolophus marthae, is one of four species of iguanas (three terrestrial and one marine) in the Galápagos Islands, and the only one listed as critically endangered by the IUCN. The species can only be found on the north-west slopes of the highest volcano on Isabela Island and was first described to science in 2009. As part of a population telemetry study, a health assessment was authorized by the Galápagos National Park. Wild adult iguanas were captured on Wolf Volcano in September 2019 and April 2021 to record morphological and physiological parameters including body temperature, heart rate, intraocular pressures, tear formation, and infrared iris images. Blood samples were also collected and analyzed. An i-STAT portable blood analyzer was used to obtain values for base excess in the extracellular fluid compartment (BEecf), glucose (Glu), hematocrit (HctPCV), hemoglobin (Hb), ionized calcium (iCa), partial pressure of carbon dioxide (pCO2), partial pressure of oxygen (pO2), percent oxygen saturation (sO2%), pH, potassium (K), and sodium (Na). When possible, data were compared to previously published and available data for the other Galápagos iguanas. The results reported here provide baseline values that will be useful in detecting changes in health status among pink land iguanas affected by climate change, invasive species, anthropogenic threats, or natural disturbances. The collected data also provide an invaluable resource for conservation scientists planning to implement conservation strategies, like translocations, that may temporarily alter these baseline values.

Limited natural regeneration of unique Scalesia forest following invasive plant removal in Galapagos.

More than 60% of the flora of the Galapagos Islands is introduced and some of these species have become invasive, severely altering ecosystems. An example of an affected ecosystem is the Scalesia forest, originally dominated by the endemic giant daisy tree Scalesia pedunculata (Asteraceae). The remnant patches of this unique forest are increasingly being invaded by introduced plants, mainly by Rubus niveus (blackberry, Rosaceae). To help large-scale restoration of this ecologically important forest, we seek to better understand the natural regeneration of S. pedunculata after invasive plant control. We monitored naturally recruited S. pedunculata saplings and young trees over five years in an area where invasive plant species are continuously being removed by manual means. We measured survival, height and growth of S. pedunculata saplings and young trees along permanent transects. Percent cover of surrounding plant species and of canopy shade directly above each S. pedunculata individual were determined, as well as distance to the next mature S. pedunculata tree. We identified potential factors influencing initial sapling survival and growth by applying generalized linear models. Results showed a rapid growth of saplings and young trees of up to 0.45 cm per day and a high mortality rate, as is typical for pioneer species like S. pedunculata. Sapling survival, growth and mortality seemed to be influenced by light availability, surrounding vegetation and distance to the next adult S. pedunculata tree. We concluded that natural regeneration of S. pedunculata was high only five months after the last herbicide application but that 95% of these recruits had died over the 5-year period. Further studies are needed to corroborate whether the number of surviving trees is sufficient to replace the aging adult trees and this way maintain remnants of the Scalesia forest. Urgent action is needed to help improve future restoration strategies to prevent further degradation of this rapidly shrinking threatened forest ecosystem.

First record of Hirsutella saussurei in the Galápagos Islands and first evidence parasitizing the invasive paper wasp, Polistes versicolor

Abstract In the present work we report the first occurrence of the entomogenous fungus Hirsutella saussurei, in the Galápagos Islands, and also the first evidence of this fungus parasitizing a new host: the invasive paper wasp Polistes versicolor. Some wasps parasitized by the fungus were found in Santa Cruz island in 2018 and 2019, while two new specimens were found in Floreana in 2019. Our data enlarge both the geographical distribution of the fungus and the host range. Even though P. versicolor is considered an important threat for the Galapagos endemic fauna, it is necessary a deep research to know if H. saussurei could be included under control programs of this invasive wasp.