Invasion by Cedrela odorata threatens long distance migration of Galapagos tortoises.

Invasive alien species are among the most pervasive threats to biodiversity. Invasive species can cause catastrophic reductions in populations of native and endemic species and the collapse of ecosystem function. A second major global conservation concern is the extirpation of large-bodied mobile animals, including long-distance migrants, which often have keystone ecological roles over extensive spatial extents. Here, we report on a potentially catastrophic synergy between these phenomena that threatens the endemic biota of the Galapagos Archipelago. We used GPS telemetry to track 140 migratory journeys by 25 Western Santa Cruz Island Galapagos tortoises. We plotted the spatial interaction between tortoise migrations and recently established non-native forest dominated by the invasive tree Cedrela odorata (Cedrela forest). We qualified (a) the proportion of migratory journeys that traversed Cedrela forest, and (b) the probability that this observed pattern occurred by chance. Tortoise migrations were overwhelmingly restricted to small corridors between Cedrela forest blocks, indicating clear avoidance of those blocks. Just eight of 140 migrations traversed extensive Cedrela stands. Tortoises avoid Cedrela forest during their migrations. Further expansion of Cedrela forest threatens long-distance migration and population viability of critically endangered Galapagos tortoises. Applied research to determine effective management solutions to mitigate Cedrela invasion is a high priority.

Temperature along an elevation gradient determines Galapagos tortoise sex ratios.

Climate change threatens endemic island ectothermic reptiles that display small population sizes and temperature-dependent sex determination (TSD). Studies of captive Galapagos tortoises demonstrate type A TSD with warmer incubation temperatures producing females. However, there are few published data from free-living Galapagos tortoises on incubation temperature regimes, and none on hatchling sex ratios in the wild or the potential impacts of climate change on future sex ratios. We sought to address these deficits by quantifying incubation temperatures of nests and sex ratios of juvenile tortoises along an elevation gradient on Santa Cruz Island. We focused on three geographically separated nesting zones with mean elevations of 14 m (lower), 57 m (middle), and 107 m (upper) above sea level. Nest temperatures in 54 nests distributed across the three nesting zones were measured every 4 h throughout the incubation period using iButton thermochrons. We used coelioscopy to conduct visual exams of gonads to determine the sex of 40 juvenile tortoises from the three nesting zones. During the middle trimester of incubation, the period during which sex is determined in turtles, mean nest temperatures were 25.75°C (SD = 1.08) in the upper zone, and 27.02°C (SD = 1.09), and 27.09°C (SD = 0.85) in the middle and lower zones, respectively. The proportion of juveniles that was male increased from 11.1% in the lower zone and 9.5% in the middle zone, to 80% in the upper zone. A ca. 50 m increase in elevation induced a decrease of >1.25°C in mean nest temperature during the second trimester of incubation. Over the same elevation change, the proportion of males in the juvenile tortoise population increased by ca. 70%. Temperatures on Galapagos are predicted to increase by 1-4°C over the next 50 years, which is likely to increase the frequency of female tortoises across the archipelago.

Antimicrobial resistance in Galapagos tortoises as an indicator of the growing human footprint.

Antimicrobial resistance has become one of the main public health threats worldwide with anthropogenic activities driving the spread of resistance. Understanding and combatting the spread of resistant bacteria is a top priority for global health institutions, and it is included as one of the main goals of the One Health initiative. Giant tortoises (Chelonoidis spp.), some of the most iconic species on Earth, are widely distributed across the Galapagos archipelago and are thus perfect candidates to test the hypothesis that wildlife species in the Galapagos carry antimicrobial resistant genes (ARGs) associated with human activities. We sampled a total of 200 free-living Galapagos tortoises from western Santa Cruz Island (C. porteri), the most human-populated island of the archipelago, and 70 tortoises (C. vandenburghi) from the isolated Alcedo Volcano on Isabela Island, a natural area with minimal human presence. Fecal samples were analyzed by quantitative PCR for a panel of 21 ARGs conferring resistance for eight antimicrobial classes. We found ARGs in both Santa Cruz and Alcedo Volcano giant tortoises; however, both qualitative and quantitative results showed higher loads of ARGs in tortoises inhabiting the human modified environments of Santa Cruz. Moreover, Santa Cruz tortoises sampled in higher human-modified landscapes (i.e., farmlands and urban areas) presented a higher number of ARGs, antimicrobial classes, and multi-resistant microbiomes than those from less anthropized areas within the same island. Our findings suggest that human activities in Galapagos have a negative impact on ecosystem health through ARG dispersal. This research highlights a new threat for the health and conservation of the unique wildlife of the Galapagos, their ecosystems, and the humans inhabiting this World Heritage Site. Our recommendation to local policy makers is to control and reduce the use of antibiotics in both human and animal health, thus helping enforce antimicrobial regulations.

FIELD ANESTHESIA AND GONADAL MORPHOLOGY OF IMMATURE WESTERN SANTA CRUZ TORTOISES (CHELONOIDIS PORTERI).

Evaluation of sex ratios is a critical component of chelonian captive breeding programs and may become increasingly useful to assess the demographics of free-living populations. In many reptile species, the sex of immature animals cannot be determined based on external features. Endoscopic sex identification is an accurate and safe method to identify the sex of immature individuals of some chelonian species. A number of studies describe this technique in controlled, hospital settings and report significant interspecies variations in gonad morphology; however, there are few reports describing this technique in field conditions. In the current study, the gonadal morphology of 40 immature Western Santa Cruz tortoises (Chelonoidis porteri) on Santa Cruz Island in Galapagos, Ecuador, was assessed. A previously described endoscopic protocol was used to perform sex identification under field conditions. Tortoises were anesthetized using an intramuscular injection of ketamine (10 mg/kg) and medetomidine (0.1 mg/kg), which provided an adequate plane of anesthesia. The medetomidine was reversed with atipamezole (0.5 mg/kg). Field conditions presented challenges such as limited control over lighting, suboptimal patient positioning, and restricted power supply for endoscopy equipment. The immature testicle in Western Santa Cruz tortoises was oval, reddish pink, and tightly adhered to the coelomic membrane ventral to the kidney. The surface of the gonads resembled other species with the notable exception that the ovaries lacked a significant number of primordial follicles. These gonadal characteristics were consistent, with only one individual identified as undetermined sex of the 40 samples. This field-based endoscopic gonadal evaluation was a safe and sensitive technique for determining the sex of free-living immature Western Santa Cruz Galapagos tortoises.

Migration triggers in a large herbivore: Galápagos giant tortoises navigating resource gradients on volcanoes.

To understand how migratory behavior evolved and to predict how migratory species will respond to global environmental change it is important to quantify the fitness consequences of intra- and inter-individual variation in migratory behavior. Intra-individual variation includes behavioral responses to changing environmental conditions and hence behavioral plasticity in the context of novel or variable conditions. Inter-individual variation determines the degree of variation on which selection can act and the rate of evolutionary responses to changes in average and extreme environmental conditions. Here we focus on variation in the partial migratory behavior of giant Galápagos tortoises (Chelonoidis spp.) and its energetic consequences. We evaluate the extent and mechanisms by which tortoises adjust migration timing in response to varying annual environmental conditions, and integrate movement data within a bioenergetic model of tortoise migration to quantify the fitness consequences of migration timing. We find strong inter-individual variation in the timing of migration, which was not affected by environmental conditions prevailing at the time of migration but rather by average expectations estimated from multi-annual averaged conditions. This variation is associated with an average annual loss in efficiency of ~15% relative to optimal timing based on year-specific conditions. These results point towards a limited ability of tortoises to adjust the timing of their migrations based on prevailing (and, by extension, future) conditions, suggesting that the adaptability of tortoise migratory behavior to changing conditions is predicated more by past "normal" conditions than responses to prevailing, changing conditions. Our work offers insights into the level of environmental-tuning in migratory behavior and a general framework for future research across taxa.

Allometric and temporal scaling of movement characteristics in Galapagos tortoises.

Understanding how individual movement scales with body size is of fundamental importance in predicting ecological relationships for diverse species. One-dimensional movement metrics scale consistently with body size yet vary over different temporal scales. Knowing how temporal scale influences the relationship between animal body size and movement would better inform hypotheses about the efficiency of foraging behaviour, the ontogeny of energy budgets, and numerous life-history trade-offs. We investigated how the temporal scaling of allometric patterns in movement varies over the course of a year, specifically during periods of motivated (directional and fast movement) and unmotivated (stationary and tortuous movement) behaviour. We focused on a recently diverged group of species that displays wide variation in movement behaviour - giant Galapagos tortoises (Chelonoidis spp.) - to test how movement metrics estimated on a monthly basis scaled with body size. We used state-space modelling to estimate seven different movement metrics of Galapagos tortoises. We used log-log regression of the power law to evaluate allometric scaling for these movement metrics and contrasted relationships by species and sex. Allometric scaling of movement was more apparent during motivated periods of movement. During this period, allometry was revealed at multiple temporal intervals (hourly, daily and monthly), with values observed at daily and monthly intervals corresponding most closely to the expected one-fourth scaling coefficient, albeit with wide credible intervals. We further detected differences in the magnitude of scaling among taxa uncoupled from observed differences in the temporal structuring of their movement rates. Our results indicate that the definition of temporal scales is fundamental to the detection of allometry of movement and should be given more attention in movement studies. Our approach not only provides new conceptual insights into temporal attributes in one-dimensional scaling of movement, but also generates valuable insights into the movement ecology of iconic yet poorly understood Galapagos giant tortoises.