The Gobiidae family occupy one of the most diverse habitat ranges of all fishes. One key reason for their successful colonization of different habitats is their ability to adapt to different energy demands. This energy requirement is related to the ability of mitochondria in cells to generate energy via oxidative phosphorylation (OXPHOS). Here, we assembled three complete mitochondrial genomes of Rhinogobius shennongensis, Rhinogobius wuyanlingensis, and Chaenogobius annularis. These mitogenomes are circular and include 13 protein-coding genes (PCGs), two rRNAs, 22 tRNAs, and one non-coding control region (CR). We used comparative mitochondrial DNA (mtDNA) genome and selection pressure analyses to explore the structure and evolutionary rates of Gobiidae mitogenomics in different environments. The CmC model showed that the ω ratios of all mtDNA PCGs were <1, and that the evolutionary rate of adenosine triphosphate 8 (atp8) was faster in Gobiidae than in other mitochondrial DNA PCGs. We also found evidence of positive selection for several sites of NADH dehydrogenase (nd) 6 and atp8 genes. Thus, divergent mechanisms appear to underlie the evolution of mtDNA PCGs, which might explain the ability of Gobiidae to adapt to diverse environments. Our study provides new insights on the adaptive evolution of Gobiidae mtDNA genome and molecular mechanisms of OXPHOS.

The wolf (Canis lupus) is one of the most widely distributed terrestrial mammals, because it is well adapted to various ecological niches and their corresponding pathogen environments. Immunological competence is a crucial factor involved in adapting to a changing environment and fighting pathogen infection in animals. In this study, the peripheral blood transcriptome of wolves was generated via RNA-seq to advance understanding of the wolf immunome, with a special focus on the major histocompatibility complex class I (MHC I) and toll-like receptor (TLR) gene families, which are involved in pathogen recognition and defense. The blood transcriptomic libraries of eight wolves originating from Tibet and Inner Mongolia were sequenced, and approximately 383 million reads were generated. Using a genome-guided assembly strategy, we obtained 123,851 unigenes, with a mean length of 845 bp and an N50 length of 1121 bp. On the basis of BLAST searches against the NCBI non-redundant protein database (Nr), a total of 36,192 (29.22%) unigenes were annotated. For functional classification, 24,663 unigenes were assigned to 13,016 Gene Ontology (GO) terms belonging to 51 sub-categories of the three main GO categories. Additionally, 7682 unigenes were classified into 6 Kyoto Encyclopedia of Genes and Genomes (KEGG) categories, in which the most represented functional sub-categories were signal transduction and the immune system, and 16,238 unigenes were functionally classified into 25 Eukaryotic Orthologous Groups (KOG) categories. We observed an overall higher ω (d N/d S) value at antigen-binding sites (ABSs) than at non-ABS regions as well as clear evidence of intergenic/intragenic recombination events at wolf MHC I loci. Additionally, our analysis revealed that carnivorous TLRs were dominated by purifying selection, with mean ω values at each TLR locus ranging from 0.173 to 0.527. However, we also found significant instances of positive selection that acted on several codons in pathogen recognition domains and were linked to species-specific differences in pathogen recognition. This study represents the first attempt to characterize the blood transcriptome of the wolf and to highlight the value of investigating the immune system. Balancing selection and recombination have contributed to the historical evolution of wolf MHC I genes. Moreover, TLRs in carnivores have undergone adaptive evolution against the background of purifying selection, and a high level of adaptive evolution was detected in the wolf TLR system.

P. brasiliensis and L. canadensis are two otter species, which successfully occupied semi-aquatic habitats and diverged from other Mustelidae. Herein, the full-length mitochondrial genome sequences were constructed for these two otter species for the first time. Comparative mitochondrial genome, selection pressure, and phylogenetic independent contrasts (PICs) analyses were conducted to determine the structure and evolutionary characteristics of their mitochondrial genomes. Phylogenetic analyses were also conducted to confirm these two otter species’ phylogenetic position. The results demonstrated that the mitochondrial genome structure of P. brasiliensis and L. canadensis were consistent across Mustelidae. However, selection pressure analyses demonstrated that the evolutionary rates of mitochondrial genome protein-coding genes (PCGs) ND1, ND4, and ND4L were higher in otters than in terrestrial Mustelidae, whereas the evolutionary rates of ND2, ND6, and COX1 were lower in otters. Additionally, PIC analysis demonstrated that the evolutionary rates of ND2, ND4, and ND4L markedly correlated with a niche type. Phylogenetic analysis showed that P. brasiliensis is situated at the base of the evolutionary tree of otters, and then L. canadensis diverged from it. This study suggests a divergent evolutionary pattern of Mustelidae mitochondrial genome PCGs, prompting the otters’ adaptation to semi-aquatic habitats.

Host phylogeny can imprint the gut microbiota, but it is often masked by diet and environment. Leveraging the standardized husbandry of a zoological collection, we profiled fecal microbiota from 55 captive artiodactyls representing 12 species in Bovidae, Cervidae, and Camelidae using 16S rRNA amplicon sequencing targeting the V3–V4 region on the Illumina MiSeq platform. Community composition differed significantly among host families (Bray–Curtis PERMANOVA, R² = 0.1075, p = 0.001). A host–microbiota tanglegram, which juxtaposes the host phylogeny with a dendrogram of microbiota similarity, recovered a topology congruent with the host phylogeny, with camelids forming a distinct branch separate from true ruminants in both trees. The linear discriminant analysis effect size (LEfSe; LDA ≥ 3.5) identified family-specific biomarkers, including enrichment of Acinetobacter/Moraxellaceae in Bovidae, Rikenellaceae (the Rikenellaceae_RC9_gut_group) in Cervidae, and Rummeliibacillus together with the Christensenellaceae_R-7_group in Camelidae. Functional inference with PICRUSt2 revealed significant differences in KEGG level-2 pathways (e.g., carbohydrate metabolism and xenobiotics biodegradation), consistent with taxonomic shifts. Altogether, these findings show that—even under uniform captive conditions—host evolutionary history remains a primary determinant of both the structure and the predicted functions of the artiodactyl gut microbiota, refining the scope of phylosymbiosis and providing actionable baselines for veterinary monitoring and conservation management in zoo settings.

Bat groups have a high degree of species diversity, and the taxonomic status and phylogenetic relationships among bat species have always been research hotspots. Due to the fact that morphological characteristics do not always reflect the evolutionary relationships among species, mitochondrial DNA has been widely used in the study of species relationships due to its maternal inheritance pattern. Myotis aurascens has been suggested as a possible synonym for M. davidii. However, the status of this classification has been controversial. In this study, the morphological and molecular characteristics of a M. aurascens captured from Inner Mongolia, China, were analyzed to determine its taxonomic status. In terms of morphological features, the body weight was 6.33 g, the head and body length were 45.10 mm, the forearm length was 35.87 mm, and the tragus length was 7.51 mm. These values all fell within the species signature data range. Nucleotide skew analysis of the protein-coding genes (PCGs) suggested that only five PCGs (ND1, ND2, COX2, ATP8, and ND4) showed AT-skew value within the mitogenome of M. aurascens. Except for ND6, the GC-skew values of the other PCGs were negative, reflecting the preference for C and T bases compared to G and A bases. Molecular phylogenetic analyses based on mitochondrial PCGs indicated that M. aurascens was a distinct species from M. davidii and phylogenetically closer to M. ikonnikovi, M. alcathoe, and M. mystacinus. Genetic distance analysis also showed that M. aurascens and M. davidii were distantly related. Therefore, the integrated analysis demonstrated that M. aurascens should be considered a distinct species rather than a synonym of M. davidii. Our study could provide a reference for enriching species diversity and research on conservation in China.

Black-billed capercaillie (Tetrao parvirostris) was listed as a first-class state-protected animal because it was endangered in China (Category I). This study is the first to examine the diversity and composition of T. parvirostris gut microbiome in the wild. We collected fecal samples from five black-billed capercaillie flock roosting sites (each 20 km apart) in one day. Thirty fecal samples were sequenced with 16S rRNA gene amplicons on the Illumina HiSeq platform. This study is the first to analyze the fecal microbiome composition and diversity of black-billed capercaillie in the wild. At the phylum level, Camplyobacterota, Bacillota, Cyanobacteria, Actinomycetota, and Bacteroidota were the most abundant in the fecal microbiome of black-billed capercaillie. At the genus level, unidentified Chloroplast, Escherichia−Shigella, Faecalitalea, Bifidobacterium, and Halomonas were the dominant genera. Based on alpha and beta diversity analyses, we found no significant differences in the fecal microbiome between five flocks of black-billed capercaillie. Protein families: genetic information processing; protein families: signaling and cellular processes, carbohydrate metabolism; protein families: metabolism and energy metabolism are the main predicted functions of the black-billed capercaillie gut microbiome through the PICRUSt2 method. This study reveals the composition and structure of the fecal microbiome of the black-billed capercaillie under wild survival conditions, and this study provides scientific data for the comprehensive conservation of the black-billed capercaillie.