Assessing Marine Endocrine-Disrupting Chemicals in the Critically Endangered California Condor: Implications for Reintroduction to Coastal Environments.

Coastal reintroduction sites for California condors (Gymnogyps californianus) can lead to elevated halogenated organic compound (HOC) exposure and potential health impacts due to the consumption of scavenged marine mammals. Using nontargeted analysis based on comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC×GC/TOF-MS), we compared HOC profiles of plasma from inland and coastal scavenging California condors from the state of California (CA), and marine mammal blubber from CA and the Gulf of California off Baja California (BC), Mexico. We detected more HOCs in coastal condors (32 ± 5, mean number of HOCs ± SD, n = 7) than in inland condors (8 ± 1, n = 10) and in CA marine mammals (136 ± 87, n = 25) than in BC marine mammals (55 ± 46, n = 8). ∑DDT-related compounds, ∑PCBs, and total tris(chlorophenyl)methane (∑TCPM) were, respectively, ∼7, ∼3.5, and ∼148 times more abundant in CA than in BC marine mammals. The endocrine-disrupting potential of selected polychlorinated biphenyls (PCB) congeners, TCPM, and TCPMOH was determined by in vitro California condor estrogen receptor (ER) activation. The higher levels of HOCs in coastal condors compared to those in inland condors and lower levels of HOC contamination in Baja California marine mammals compared to those from the state of California are factors to consider in condor reintroduction efforts.

Facultative Parthenogenesis in California Condors.

Parthenogenesis is a relatively rare event in birds, documented in unfertilized eggs from columbid, galliform, and passerine females with no access to males. In the critically endangered California condor, parentage analysis conducted utilizing polymorphic microsatellite loci has identified two instances of parthenogenetic development from the eggs of two females in the captive breeding program, each continuously housed with a reproductively capable male with whom they had produced offspring. Paternal genetic contribution to the two chicks was excluded. Both parthenotes possessed the expected male ZZ sex chromosomes and were homozygous for all evaluated markers inherited from their dams. These findings represent the first molecular marker-based identification of facultative parthenogenesis in an avian species, notably of females in regular contact with fertile males, and add to the phylogenetic breadth of vertebrate taxa documented to have reproduced via asexual reproduction.

Identification of California condor (Gymnogyps californianus) estrogen receptor variants and their activation by xenoestrogens.

California condors released in costal sites are exposed to high levels of xenoestrogens, particularly p,p'-DDE, through scavenging of marine mammal carcasses. As a result, coastal condors carry a higher contaminant loads and experience eggshell thinning when compared to their inland counterparts. Given that condor estrogen receptors (Esrs) are activated by physiologically relevant levels of xenoestrogens, differences in vulnerability to endocrine disruption may exist depending on which Esr variant(s) an individual condor possesses. This work aims to characterize genetic polymorphisms in estrogen receptor genes (ESRs) in California condors; one identified for condor estrogen receptor 1 (ESR1) (N161S, E162D) and one in the ESR2 (T114M) gene. Each variant was confirmed in individual founder birds by direct PCR sequencing as well as in first generation offspring to understand the introduction of the alleles into the pedigree (6 birds for ESR1 and 5 birds for ESR2). Site-directed mutagenesis was performed on wild type receptors to produce each of the full-length ESR variants and activation of Esr1 and Esr2 variant and wild type receptors by xenoestrogens was compared. Maximal activation of the variant form of Esr1 was significantly higher (p < 0.05) in response to ethinyl estradiol (EE2), o,p'-DDE, p,p'-DDE, p,p'-DDT and p,p'-DDD compared to wild type Esr1. For Esr2 the wild type maximal activation was higher in response to o,p'-DDE, p,p'-DDE, o,p'-DDT, and p,p'-DDT. Although significant differences in activation of condor Esr variants by xenoestrogens occurred at high (micromolar) concentrations, they correspond to circulating concentrations previously reported in coastal birds. Release and relocation of California condors to the coast is a promising avenue for recovery, however, reproductive problems associated with xenoestrogen exposure pose a sub-lethal threat to long-term success. Based on above findings, future release decisions could be informed by ESR form(s) individual birds possess to reduce deleterious effects of xenoestrogen exposure and ultimately improve reproductive success in wild populations.

ANALYSIS OF HISTORICAL MEDICAL RECORDS OF CALIFORNIA CONDORS ( GYMNOGYPS CALIFORNIANUS) ADMITTED FOR LEAD EXPOSURE TO THE LOS ANGELES ZOO AND BOTANICAL GARDENS BETWEEN 1997 AND 2012: A CASE SERIES STUDY.

Lead toxicosis remains the primary cause of death in wild populations of California condors ( Gymnogyps californianus). Many condors require medical intervention with lead chelation therapy, among other conservation measures, to survive in the wild. An analysis of historical medical records of California condors admitted for lead exposure to the Los Angeles Zoo and Botanical Gardens (LA Zoo) between 1997 and 2012 was performed to investigate clinical presentation, radiographic findings, and treatment along with the potential impact of these factors on recovery and successful release back into the wild. Of 100 individual condors presented to the LA Zoo for suspected lead toxicosis during this period, 84 condors had records for initial laboratory blood lead levels. For these 84 condors (40 [47.6%] females and 44 [52.4%] males), 277 initial laboratory blood lead levels in total were recorded because of repeated admissions. Thirty-two (38%) condors were admitted once to the zoo and 62% were admitted two or more times. Clinical signs were not observed in 87.5% of the condors admitted with median blood lead concentrations of 26 µg/dl. Of the radiographs taken on initial presentation, 65% did not show evidence of gastrointestinal metallic foreign bodies. Various treatments protocols with edetatum calcium disodium, dimercaptosuccinic acid, and lactated Ringer's solution were documented in the medical records. Of the 277 admissions, 154 admissions had a recorded outcome posttreatment of which 140 (91%) admissions were released back to the wild. This study contributes to a better understanding of how intervention and therapeutic approaches have been essential for the recovery, release, and sustainability of these birds as a wild population. Consistency in data collection of California condors admitted for diagnosis and treatment of lead toxicosis is needed to better assess the impacts of medical interventions.

Linking cases of illegal shootings of the endangered California condor using stable lead isotope analysis.

Lead poisoning is preventing the recovery of the critically endangered California condor (Gymnogyps californianus) and lead isotope analyses have demonstrated that ingestion of spent lead ammunition is the principal source of lead poisoning in condors. Over an 8 month period in 2009, three lead-poisoned condors were independently presented with birdshot embedded in their tissues, evidencing they had been shot. No information connecting these illegal shooting events existed and the timing of the shooting(s) was unknown. Using lead concentration and stable lead isotope analyses of feathers, blood, and recovered birdshot, we observed that: i) lead isotope ratios of embedded shot from all three birds were measurably indistinguishable from each other, suggesting a common source; ii) lead exposure histories re-constructed from feather analysis suggested that the shooting(s) occurred within the same timeframe; and iii) two of the three condors were lead poisoned from a lead source isotopically indistinguishable from the embedded birdshot, implicating ingestion of this type of birdshot as the source of poisoning. One of the condors was subsequently lead poisoned the following year from ingestion of a lead buckshot (blood lead 556 µg/dL), illustrating that ingested shot possess a substantially greater lead poisoning risk compared to embedded shot retained in tissue (blood lead ~20 µg/dL). To our knowledge, this is the first study to use lead isotopes as a tool to retrospectively link wildlife shooting events.

Supervised versus unsupervised approaches to classification of accelerometry data.

Sophisticated animal-borne sensor systems are increasingly providing novel insight into how animals behave and move. Despite their widespread use in ecology, the diversity and expanding quality and quantity of data they produce have created a need for robust analytical methods for biological interpretation. Machine learning tools are often used to meet this need. However, their relative effectiveness is not well known and, in the case of unsupervised tools, given that they do not use validation data, their accuracy can be difficult to assess. We evaluated the effectiveness of supervised (n = 6), semi-supervised (n = 1), and unsupervised (n = 2) approaches to analyzing accelerometry data collected from critically endangered California condors (Gymnogyps californianus). Unsupervised K-means and EM (expectation-maximization) clustering approaches performed poorly, with adequate classification accuracies of <0.8 but very low values for kappa statistics (range: -0.02 to 0.06). The semi-supervised nearest mean classifier was moderately effective at classification, with an overall classification accuracy of 0.61 but effective classification only of two of the four behavioral classes. Supervised random forest (RF) and k-nearest neighbor (kNN) machine learning models were most effective at classification across all behavior types, with overall accuracies >0.81. Kappa statistics were also highest for RF and kNN, in most cases substantially greater than for other modeling approaches. Unsupervised modeling, which is commonly used for the classification of a priori-defined behaviors in telemetry data, can provide useful information but likely is instead better suited to post hoc definition of generalized behavioral states. This work also shows the potential for substantial variation in classification accuracy among different machine learning approaches and among different metrics of accuracy. As such, when analyzing biotelemetry data, best practices appear to call for the evaluation of several machine learning techniques and several measures of accuracy for each dataset under consideration.

Towards a Genetic Linkage Map of the California Condor, an Endangered New World Vulture Species.

The development of a linkage map is an important component for promoting genetic and genomic studies in California condors, an endangered New World vulture species. Using a set of designed anonymous microsatellite markers, we genotyped a reference condor population involving 121 individuals. After marker validation and genotype filtering, the genetic linkage analysis was performed using 123 microsatellite loci. This resulted in the identification of 15 linkage groups/subgroups that formed a first-generation condor genetic map, while no markers linked to a lethal chondrodystrophy mutation were found. A panel of polymorphic markers that is instrumental in molecular parentage diagnostics and other genetic studies in the California condor was selected. Further condor conservation genomics research will be focused on updating the linkage map and integrating it with cytogenetic and BAC-based physical maps and ultimately with the genome sequence assembly.

Effects of Lead Exposure, Flock Behavior, and Management Actions on the Survival of California Condors (Gymnogyps californianus).

Translocation is an increasingly important tool for managing endangered species, but factors influencing the survival of translocated individuals are not well understood. Here we examine intrinsic and extrinsic drivers of survival for critically endangered California condors (Gymnogyps californianus) whose wild population recovery is reliant upon releases of captively bred stock. We used known fate models and information-theoretic methods to compare the ability of hypothesized covariates, most of which serve as proxies for lead exposure risk, to predict survival rates of condors in California. Our best supported model included the following predictors of survival: age of the recovery program, precipitation, proportion of days observed feeding on proffered carcasses, maximum blood lead concentration over the preceding 18 months, and time since release. We found that as flocks have increased in size and age, condors are increasingly likely to range more widely and less likely to be observed feeding on proffered food, and these "wilder" behaviors were associated with lower survival. After accounting for these behaviors, we found a positive survival trend, which we attribute to ongoing improvements in management. Our findings illustrate that the survival of translocated animals, such as highly social California condors, is influenced by behaviors that change through time.

Lead Exposure Risk from Trash Ingestion by the Endangered California Condor (Gymnogyps californianus).

Lead poisoning from ingestion of spent lead ammunition is one of the greatest threats to the recovery of California Condors (Gymnogyps californianus) in the wild. Trash ingestion by condors is well documented, yet the extent that trash presents a lead exposure risk is unknown. We evaluated 1,413 trash items collected from condor nest areas and nestlings in the Transverse Range of Ventura County, California, US, from 2002 to 2008, for their potential as a lead exposure risk to condors. We visually identified 71 items suspected to contain sufficient lead to be of toxicologic concern. These items were leached with weak acid and analyzed for lead. Twenty-seven of the 71 leached items (~2% of the 1,413 items) were "lead containing" based on criteria of a leachate lead concentration >1 µg/mL, with the majority of these items (22; 81% of the 27 lead items) being ammunition related (e.g., spent bullet casings and jacketed bullets). Only three of the 1,413 items collected were lead containing but were clearly not ammunition related; the other two lead-containing items were unidentified. Our results suggest that trash ingestion of nonammunition items does not pose a significant lead exposure risk to the California Condor population in California.