Search Results (42)

Ethnobiological gardens conserve both biological and cultural diversity, but their permeable boundaries may expose native wildlife inside the garden to chronic domestic carnivore activity. In this single-garden case study, we compare small mammal assemblages between the Jardín Etnobiológico Tlaxcallan (JET), a periurban ethnobiological garden in Tlaxcala, Mexico, and its adjacent, heterogeneous Periphery of native vegetation and agricultural matrix, using live traps, camera traps, and direct observations over nine months. Across 162 detections of 9 species, the Periphery consistently showed higher richness and diversity than the JET at every Hill number order and the two assemblages were compositionally distinct (Bray–Curtis = 0.679), with the JET dominated by disturbance-tolerant species and the Periphery additionally harboring habitat-sensitive natives that were largely or entirely absent from the JET. Domestic cats were detected almost exclusively within the JET, while dogs were recorded at both sites. Dog and cat detection rates were both positively associated with species richness in generalized linear mixed models, a pattern that most plausibly reflects spatial co-occurrence between carnivores and disturbance-tolerant species within the JET rather than a facilitative effect of carnivores on richness. Because our design contrasts a single garden with a single Periphery, these results cannot separate the influence of domestic carnivores from co-varying differences in habitat structure, sampling geometry, or human activity. We therefore present the observed compositional turnover as a hypothesis-generating pattern consistent with—but not evidence of—carnivore-associated ecological filtering that warrants testing through replicated, multi-site designs before informing carnivore management in periurban ethnobiological gardens. Full article

Geographic and ecological variations are frequently associated with differences in the microbiota of arthropod vectors, with potential implications for pathogen transmission and public health. This study characterized and compared the bacterial microbiota associated with the brown dog tick (Rhipicephalus sanguineus) across three ecologically contrasting regions of Mexico: Cancun (Quintana Roo), Comarca Lagunera (Durango–Coahuila), and Hermosillo (Sonora). Non-engorged ticks collected from stray dogs were analyzed using 16S rRNA gene (V3–V4) sequencing. Amplicon sequence variants (ASVs) generated in QIIME2 were used for taxonomic, diversity, and predictive functional analyses. Proteobacteria dominated all samples, with Coxiella-like bacteria tentatively assigned as Coxiella mudrowiae identified as a dominant taxon across all localities. Significant geographic differences were observed in alpha and beta diversity, with Comarca Lagunera showing the highest diversity and Hermosillo the lowest. Sequences tentatively assigned to Rickettsia rickettsii were detected exclusively in two pools from Hermosillo. Functional predictions revealed a conserved metabolic repertoire alongside geographic variation in pathway abundance. Overall, the results support the existence of a stable symbiotic component accompanied by a geographically variable bacterial fraction associated with ecologically contrasting regions. These findings highlight the importance of geographic context in shaping tick-associated bacterial communities. Full article

Primary pulmonary adenocarcinoma in dogs is rare, and effective systemic therapies remain limited. To evaluate the molecular basis of potential precision oncology approaches, hotspot mutations in HER2, BRAF, KRAS, and PIK3CA were analyzed in 20 surgically resected canine pulmonary adenocarcinomas and three canine pulmonary adenocarcinoma cell lines. HER2 V659E and BRAF V595E mutations were each detected in 3/20 cases (15%), while KRAS G12V was detected in 1/20 cases (5%). No PIK3CA hotspot mutations were identified. The BRAF V595E mutation was additionally detected in the AZACL2 cell line. Functional analysis demonstrated increased sensitivity of AZACL2 cells to the BRAF inhibitor dabrafenib and MEK inhibitors including trametinib, compared with BRAF wild-type cell lines. These findings support MAPK pathway dependency in BRAF-mutant canine pulmonary adenocarcinoma. The mutation spectrum was broadly consistent with previous reports, suggesting a conserved molecular landscape across geographic regions. Collectively, these data identify BRAF and HER2 alterations as clinically relevant candidates for molecular diagnostics and targeted therapy in canine pulmonary adenocarcinoma. Full article

Background/Objectives: Canine papillomavirus (CPV) is an important viral pathogen associated with papillomatosis in dogs, with canine papillomavirus type 1 (CPV1) and type 2 (CPV2) among the most prevalent and clinically relevant genotypes. The L1 capsid protein is a major immunogenic antigen of papillomaviruses; however, conserved linear B-cell epitopes shared between CPV genotypes remain poorly defined. This study aimed to identify conserved cross-reactive B-cell epitopes within CPV1 and CPV2 L1 proteins and to evaluate their preliminary immunoreactivity. Methods: Conserved linear B-cell epitopes were predicted through integrated bioinformatic and structural analyses based on sequence conservation and surface accessibility. Three candidate epitopes were selected. Recombinant CPV1 and CPV2 L1 proteins were expressed in Escherichia coli (E. coli), purified, used as recombinant L1 antigens, together with BSA-conjugated synthetic epitope peptides for mouse immunization. Antigen-specific IgG responses were assessed by ELISA, antigen-associated IFN-γ responses were evaluated by ELISpot, and cross-reactive antibody recognition was assessed by Western blot. Results: Recombinant L1 proteins induced strong antigen-specific IgG responses in mice. The selected peptides induced detectable but weaker humoral responses compared with the recombinant L1 proteins. Among the three epitopes, TPSGSLV and TVVDNTR elicited antibodies that recognized both CPV1 and CPV2 L1 proteins, while the epitope VIVPKVS showed minimal or no detectable immunoreactivity. ELISpot analysis showed only modest antigen-associated IFN-γ responses, particularly in peptide-immunized groups. Conclusions: This study identified conserved cross-reactive linear B-cell epitope candidates within CPV1 and CPV2 L1 proteins and provided preliminary immunological evidence supporting their potential relevance for CPV antigen design. However, peptide-induced responses were weaker than those induced by recombinant L1 proteins, and VLP formation, antibody neutralizing activity, and protective efficacy were not evaluated. Further studies in dogs, including optimized antigen-display platforms, neutralization assays, and protection studies, are required to determine the practical value of these epitopes for CPV vaccine development. Full article

Misiones, Argentina, holds one of the largest remnants of the Atlantic Forest, with almost 1.4 million hectares of native forest, representing a critical landscape for sustainable biodiversity conservation. However, connectivity across this ecoregion is increasingly threatened by habitat conversion, landscape fragmentation, and poaching pressures that extend beyond protected area boundaries, undermining long-term sustainability of wildlife populations. Using conservation detection dogs, we located, collected, and genetically confirmed 198 scats belonging to four game species: 20 lowland tapir (Tapirus terrestris), 72 white-lipped peccary (Tayassu pecari), 55 collared peccary (Pecari tajacu), and 51 Azara’s agouti (Dasyprocta azarae). Analyses examining species-specific habitat associations emphasized the importance of extending inference beyond point locations to encompass species’ home ranges, with native forest consistently identified as a key component of habitat use. The high prevalence of scats in mosaics of human-modified habitats outside protected areas, especially along their borders, underscores the importance of managing these areas as part of a broader sustainable landscape matrix. While native forest fragments outside of protected areas may serve as important refugia supporting species persistence, their contribution to sustainable management depends on reducing poaching pressure across these landscapes. There is an urgent need to expand antipoaching efforts beyond protected areas and across the Atlantic Forest in the Green Corridor of Misiones while preventing ongoing deforestation and the expansion of monoculture plantations. Achieving sustainable wildlife management in this region will require integrated strategies that promote sustainable land use, conservation planning, and rural development. Full article

Background/Objectives: Following centuries of systematic eradication, grey wolf (Canis lupus) populations across Europe have experienced a significant recovery over recent decades, which leads to concerns regarding, among others, anthropogenic hybridization. In Greece, the genetic status of the wolf population is largely unknown to date. Here, we genetically monitor and test for wolf–dog hybridization events in a recently established wolf population in the Parnitha Protected Area, in close vicinity to the capital city of Greece. Methods: One hundred and twenty-four wolf scat samples were genotyped at 20 canine-specific autosomal microsatellite loci and compared to available reference tissue samples from wolves and free-ranging dogs. Results: A minimum of 31 unique wolf individuals were identified, structured into at least three packs. No wolf–dog hybrids were detected in the study area. To validate the accuracy of the microsatellite analysis, an ancestry informative 93-SNP panel was applied to non-invasive wolf DNA samples from the study area, confirming the absence of hybrids among them. However, a possible wolf–dog hybrid was detected among reference wolf samples collected in Northern Greece, where individuals with atypical morphological traits are observed. The estimated census population size was in accordance with concurrently obtained camera trapping data, while heterozygosity values were low. Conclusions: This research represents the first systematic effort in Greece to genetically monitor wolves recently established in a protected area. It highlights the need for targeted management strategies based on genetic data to ensure balanced long-term conservation of wolves in peri-urban areas. Full article

The Jianfengling region on southwestern Hainan Island is a sanctuary for a diverse range of wild animals. However, the exact extent of mammal species diversity and conservation status in Jianfengling remains largely unknown. Using camera-trapping data spanning from October 2020 to November 2021, this study focused on the diversity and distribution status of mammal species in Jianfengling, Hainan Tropical Rainforest National Park, China. The survey, with 41,571 camera days and 8091 independent detections, revealed 15 mammalian species belonging to 6 orders and 10 families. Among these detected species, one was categorized as Critically Endangered, and one as Near Threatened on the IUCN Red List. According to the Red List of China’s Vertebrates, one was categorized as Critically Endangered, one as Endangered, three as Vulnerable, and three as Near Threatened. Additionally, one was designated China’s national first-class key protected wildlife, and five were designated China’s national second-class key protected wildlife. Notably, populations of the Chinese pangolin (Manis pentadactyla) were confirmed to persist in the wild of Jianfengling. Occupancy modeling results showed that the occupancy of Rhesus monkey (Macaca mulatta), Hainan muntjac (Muntiacus nigripes), common palm civet (Paradoxurus hermaphroditus), and Mainland leopard cat (Prionailurus bengalensis) increased with increasing elevation. Vegetation type would only affect the occupancy of the Rhesus monkey. Normalized Difference Vegetation Index and anthropogenic activities had no obvious effect on the occupancy of mammals. In terms of the relative abundance index (RAI) of mammals captured by camera traps, the most prevalent species identified was the Asiatic brush-tailed porcupine (Atherurus macrourus), followed by the wild boar (Sus scrofa), Pallas’s squirrel (Callosciurus erythraeus), and Hainan muntjac. The monitoring also captured a significant number of images of domestic dogs and humans. These findings establish a baseline for species richness and occupancy of mammals in a typical island tropical rainforest in Hainan Tropical Rainforest National Park. We hope that our findings could be used to assess the effectiveness of conservation monitoring programs in the coming years, as species richness surveys provide an important baseline for conservation planning. Full article

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. Full article

A major goal of fecal pollution monitoring in the environment is to identify point sources of fecal contamination that may pose potential health risks due to animal- and human-specific pathogens. Ideal source tracking markers should have high host specificity and can be employed for the unambiguous identification of the host/fecal point sources. Conserved signature proteins (CSPs) are a class of unique, phylogenetically coherent indicators that are specific to a given taxon (e.g., genus or species). In this study, we report the identification and characterization of a new CSP, whose gene (designated as CSP-DV) is present in a single copy, and for whom homologs showing a high degree of sequence similarity are found only in genomes of Phocaeicola dorei and Phocaeicola vulgatus, two commensal species commonly found in the human gut and feces. We developed a qPCR method targeting this CSP gene to explore its usefulness as a human source tracking marker. We confirmed that the CSP-DV marker showed an absolute human sensitivity (100%) but some cross-reactivities in chicken, cats, dogs, rabbits, and rodents. In recreational water, the CSP-DV marker gene levels were well correlated with those of HF183, a well-validated human marker that predominantly targets the 16S rRNA gene of P. dorei, suggesting that it can be a new potential source tracking tool for human fecal contamination in specific environmental waters. In summary, our CSP-DV marker targets Phocaeicola clade-specific microbes and can provide an additional approach independent of the 16S rRNA gene to detect human sources of fecal pollution. Full article

Canine parvoviral (CPV) enteritis is frequently associated with systemic inflammatory response syndrome (SIRS), yet objective biomarkers for early detection remain limited. This study evaluated the diagnostic performance of C-reactive protein (CRP), albumin, and the CRP/albumin ratio (CAR) in 60 dogs (45 CPV-positive and 15 healthy controls). CRP and CAR were markedly higher in infected dogs, while albumin was significantly reduced (all p < 0.001). ROC analysis demonstrated excellent discrimination for SIRS, with AUC values of 0.87 for CRP and 0.86 for CAR and optimal ROC01 thresholds of 2.25 mg/dL and 1.23, respectively. The IDEXX CRP-based SIRS classification showed moderate agreement with the clinical reference (κ = 0.56), whereas a conditional inference tree and logistic regression models confirmed CRP as the most informative variable. The final age-adjusted model including the interaction age × CRP achieved superior performance (AUC = 0.93; κ = 0.84), indicating that CRP’s predictive effect varied with age. These results demonstrate that CRP and CAR are complementary, accessible biomarkers for assessing systemic inflammation in CPV infection and suggest that the IDEXX CRP cut-off (>3 mg/dL) may be conservative, as values above 2.25 mg/dL already reflect clinically relevant inflammatory activity. Full article

Misiones province covers < 1% of Argentina’s land area yet harbors > 50% of the country’s biodiversity, with a significant remnant of Atlantic Forest, a global biodiversity hotspot. Approximately 540,000 ha of this native forest is protected, with the remaining areas facing threats from ongoing land conversion, an expanding road network, and a growing rural population. A prior study incorporated noninvasive data on five carnivores into a multifaceted cost analysis to define the optimal location for a multispecies biological corridor, with the goal of enhancing landscape connectivity among protected areas. Subsequent analyses, with an updated framework, emphasized management strategies that balanced human–wildlife coexistence and habitat needs. Building on these efforts, our study applied ecological niche modeling to data located by conservation detection dogs, with genetics used to confirm species identity, and two land-use scenarios, to predict potential distributions of three game species—lowland tapir (Tapirus terrestris), white-lipped peccary (Tayassu pecari), and collared peccary (Pecari tajacu)—that are not only threatened by poaching, road mortality, and habitat loss but also serve as essential prey for carnivores. We assessed the suitability of unique and overlapping vegetation types, within and outside of protected areas, as well as within this multispecies corridor, identifying zones of high conservation concern that underscore the need for integrated planning of predators and prey. These results highlight that ensuring the long-term viability of wildlife across the heterogeneous land-use matrices of Misiones requires going beyond protected areas to promote functional connectivity, restore degraded habitats, and balance human–wildlife needs. Full article

Background/Objectives: The gene pool of the Apennine wolf is affected by admixture with domestic variants due to anthropogenic hybridisation with dogs. Genetic monitoring at the population level involves assessing the extent of admixture in single individuals, ranging from pure wolves to recent hybrids or wolf backcrosses, through the analysis of nuclear and mitochondrial DNA (mtDNA) markers. Although individually non-diagnostic, mtDNA is nevertheless essential for completing the final diagnosis of genetic admixture. Typically, the identification of wolf mtDNA haplotypes is carried out via sequencing of coding genes and non-coding DNA stretches. Our objective was to develop a fast real-time PCR assay to detect the mtDNA haplotypes that occur exclusively in the Apennine wolf population, as a valuable alternative to the demanding sequence-based typing. Methods: We validated a qualitative duplex real-time PCR that exploits the combined presence of diagnostic point mutations in two mtDNA segments, the NDH-4 gene and the control region, and is performed in a single-tube step through TaqMan-MGB chemistry. The aim was to detect mtDNA multi-fragment haplotypes that are exclusive to the Apennine wolf, bypassing sequencing. Results: Basic validation of 149 field samples, consisting of pure Apennine wolves, dogs, wolf × dog hybrids, and Dinaric wolves, showed that the assay is highly specific and sensitive, with genomic DNA amounts as low as 10⁻⁵ ng still producing positive results. It also proved high repeatability and reproducibility, thereby enabling reliable high-throughput testing. Conclusions: The results indicate that the assay presented here provides a valuable alternative method to the time- and cost-consuming sequencing procedure to reliably diagnose the maternal lineage of the still-threatened Apennine wolf, and it covers a wide range of applications, from scientific research to conservation, diagnostics, and forensics. Full article

Background: Canine oral melanoma (COM) is an aggressive and often fatal neoplasm in dogs, with clinical and molecular similarities to human melanoma. Despite its relevance as a comparative oncology model, the molecular mechanisms underlying COM remain poorly understood. This study aimed to characterize gene expression profiles in COM to identify differentially expressed genes (DEGs), potential biomarkers, and therapeutic targets. Methods: In this pilot study, we performed RNA sequencing (RNA-seq) on tumor and healthy oral tissue samples from dogs. Two independent analytical pipelines—Bowtie2-DESeq2 and HISAT-StringTie-Ballgown—were used to ensure robustness in DEG detection. We also conducted pathway enrichment and isoform-level analyses to investigate biological processes and alternative splicing events. Results: Both approaches identified a core set of 929 common DEGs. Key oncogenic pathways, including MAPK/ERK and cell cycle regulation, were significantly affected, with notable upregulation of BRAF, NRAS, CDK4, and MITF (log2FC = 2.86, p < 0.001). The transcription factor SOX10 and the cytokine IL-33, both previously implicated in melanoma progression, were consistently overexpressed. Additionally, NF1, a known RAS pathway inhibitor, was also upregulated. Isoform analysis revealed novel transcript variants, suggesting a complex layer of post-transcriptional regulation in COM. Many DEGs remained uncharacterized, and chromosomal distribution analysis highlighted potential genomic influences. Conclusions: Our findings provide new insights into the molecular landscape of COM, reinforcing its utility as a model for human melanoma. The identification of conserved oncogenic pathways and novel transcript variants opens avenues for further functional studies and the development of targeted therapies in both veterinary and human oncology. Full article

Canine papillomavirus (CPV) infection leads to a range of clinical manifestations from benign warts to malignant tumors in dogs, posing significant challenges in veterinary medicine due to its diverse genotypic spectrum. This study introduced broad-range and robust polymerase chain reaction (PCR) assays designed to enhance the detection, identification, and quantification of multiple CPV genotypes. By using both universal and genotype-specific primers, this protocol significantly improved diagnostic specificity and sensitivity across the 23 known CPV genotypes compared to previously described ones. The primers were designed to target conserved regions across all genotypes for general detection, as well as specific regions in the predominant genotypes CPV1 and CPV2 for detailed analysis. Validation of this protocol using synthetic plasmids and clinical samples confirmed its enhanced performance over traditional methods, as demonstrated by higher specificity and sensitivity. Additionally, the application of this PCR approach in a series of epidemiological studies provided novel insights into the distribution and prevalence of CPV genotypes, highlighting its potential utility in shaping targeted vaccination and clinical management strategies. Furthermore, the quantitative capability of this established protocol allowed for monitoring viral loads in clinical cases, offering a valuable tool for assessing treatment efficacy and disease progression. Further validation through larger-scale clinical studies will be crucial to substantiate the diagnostic accuracy and epidemiological value of the assays. Full article

Canine distemper virus (CDV) is the causative agent of a widespread infectious disease affecting both domestic and wild carnivores. Owing to its ability to cross species barriers and its high fatality rate in unvaccinated animals, CDV poses a significant conservation threat to endangered wildlife worldwide. To date, two distinct CDV lineages have been reported in Colombia, with cases documented separately in domestic dogs and wild peri-urban carnivores. This study aimed to detect and characterize the concurrent circulation of CDV in naturally infected domestic dogs and crab-eating foxes (Cerdocyon thous) from the same area in Colombia. Through molecular and phylogenetic analyses, we identified the South America/North America-4 lineage infecting both populations simultaneously. Our findings revealed high genetic variability, multiple virus reintroductions, and a close relationship with CDV strains previously detected in the United States. These results confirm the simultaneous circulation of CDV in the domestic and wildlife interface and underscore the urgent need for an integrated approach to CDV prevention and control involving both domestic and wildlife health interventions. Full article