Rapidly changing environments are contributing to the spread of non‐native species and their associated pathogens into new and vulnerable ecosystems, such as the Galapagos archipelago. These pathogens represent a significant threat to emblematic species. The Galapagos sea lion ( Zalophus wollebaeki ) (GSL) is an endangered and endemic pinniped that is increasingly at risk of acquiring infectious diseases due to interactions with introduced companion animals. Previously, we reported the first detection of antigens from Dirofilaria immitis , the parasite that causes canine heartworm disease, in the GSL. To investigate further, we developed a multifilarial PCR assay and successfully detected DNA from D. immitis and the closely related Dirofilaria repens in 10.7% of our sample cohort of juvenile GSLs. This assay, based on a conserved region in the filarial 28S gene, can be used in conjunction with restriction endonuclease digestion or Sanger sequencing to identify the species of the causative nematode. Our method proved effective without nonspecific amplification in a wide host range, and highly sensitive, detecting as little as one parasite. Further, this assay can be used in cases of immature, low‐worm burden, or all‐male infections. Our molecular approach offers a sensitive and specific method for detecting filarial parasites in wild animals. Further investigations are necessary to confirm the pathology of filarial nematodes in the GSL and their prevalence in the general population. Our identification of Dirofilarial species in the GSL underscores the urgent need for measures to manage the risk of pathogen transmission from introduced species to native wildlife.

Spontaneous canine prostate cancer (PC) is widely considered a pertinent clinical model for the human disease. While over 95% of PC in men are adenocarcinomas, arising from prostatic glandular epithelium, it is increasingly recognised that many canine PC are of urothelial origin, arising within the prostatic urethra or ducts, or through invasion from a primary urinary bladder tumour. At diagnosis, canine prostatic tumours are often poorly differentiated and widely disseminated, masking the primary site and limiting the sensitivity of cellular biomarkers. Consequently, published studies of canine PC show varying representation of glandular versus urothelial tumours, yielding conflicting observations regarding their molecular pathogenesis and clinical behaviour. We characterised DNA sequence mutations and copy number aberrations in 31 canine PC, seeking evidence supporting relevance as a disease model. Only three tumours resembled adenocarcinomas. The remainder were either histologically consistent with urothelial carcinoma ( n = 15), showed mixed glandular and urothelial morphology ( n = 4), or were carcinomas of undetermined cell type ( n = 9). BRAF V588E mutation was detected in 87% of tumours, including all three adenocarcinomas. Urinary bladder involvement was evident in 46% of cases, but none of the adenocarcinomas. Genome‐wide DNA copy number instability was apparent throughout the cohort, with chromosome 36 gain significantly associated with urothelial tumours. Hallmark alterations of human PC, such as defects within PI3K and androgen receptor signalling pathways, were not detected. Improved molecular subclassification of canine PC is needed to direct selection of relevant cases for modelling the human disease and to ensure appropriate extrapolation between canine and human studies.

Background/Objectives: Human-driven environmental change can promote cancer development in wild species, yet the pathophysiology of wildlife cancers remain largely unexplored. Urogenital carcinoma (UGC) in the California sea lion (CSL) (Zalophus californianus) is one of the most common cancer types documented in any wild mammal. The pathogenesis of UGC in CSLs is known to be multifactorial, with links to environmental contaminant exposure and infection by Otarine Herpesvirus-1 (OtHV-1); however, the genomic features of these cancers have not been thoroughly explored. Understanding UGC pathogenesis in the CSL has important implications for the health of humans and other species that share environment and diet. Methods: We leveraged the evolutionary conservation between the domestic dog and CSL genomes to perform cross-species whole-exome sequencing (WES) of CSL UGC tumors and matched normal tissue pairs. We also used PCR and Sanger sequencing to investigate the prevalence of DNA from OtHV-1. Results: Bioinformatic analyses identified shared somatic variants and DNA copy number aberrations in UGC tumor samples, including recurrent exonic single-nucleotide variants in CD274/PD-L1, and recurrent copy number gains in CD274/PD-L1, TNFRSF14, CD200, CDK4, and PLCG2. In an extended cohort of 70 CSLs (tumor, matched normals, and controls), a recurrent C > T single-nucleotide variant in exon 4 of CD274/PD-L1 was identified in 54 of 68 (79.4%) CSLs with diagnosed UGC. OtHV-1 DNA was detected in 67 of 70 individuals (95.8%). Conclusions: These results demonstrate that cross-species exome capture provides a means to identify genomic alterations that may play a role in the molecular pathogenesis of UGC in the CSL and adds to the body of evidence for an association between OtHV-1 and UGC in this species.

Background/Objectives: Anthropogenic pressures increasingly fragment populations, reduce connectivity in large mammals, and threaten genetic diversity. Genetic diversity is essential for a population’s ability to respond to environmental change and the long-term persistence of any species. The American black bear (Ursus americanus) is an important ecological regulator of many ecosystems. North Carolina (NC) is home to a large number of black bears that contribute to ecosystem function. In this study, we examined how rapidly changing environmental and anthropogenic effects influence the structure and genetic health of black bears in North Carolina. Methods: Whole genome sequencing data were generated from 82 black bears selected from populations representing two of the three state recognized management units, the Mountain Bear Management Unit (MBMU, n = 49) and the Coastal Bear Management Unit (CBMU, n = 27). Of the 82 black bears, 76 were retained after quality and relatedness filtering. These whole genome sequence data were used to identify almost 900,000 single nucleotide polymorphisms (SNPs) for population genetic analyses. Results: The data identified three genetic clusters, one within the CBMU and two within the MBMU, indicating regional substructure across the sampled populations. Compared with the MBMU, the CBMU exhibited significantly lower genetic diversity and higher inbreeding (p < 2.2 × 10⁻¹⁶), suggesting reduced connectivity and increased genetic vulnerability in the coastal population. Finally, a candidate panel of 4990 SNPs was developed that can be used for the population and individual assignment of black bears. Conclusions: These findings provide a valuable genome-wide resource for black bear conservation in North Carolina and support management strategies aimed at maintaining connectivity, preserving genetic diversity, and promoting long-term population resilience.