Chao Yang

Background: To conserve wild populations and ensure a sustainable supply of musk, China initiated the captive breeding of forest musk deer. The temporal dynamics of gut antibiotic resistance gene (ARG) profiles in captive forest musk deer along a breeding duration gradient remain poorly characterized. Methods: In this study, we employed metagenomic sequencing to systematically characterize the profiles and potential mobility of ARGs. Samples were divided into short-term, medium-term and long-term groups according to breeding durations. Results: A total of 331 ARG subtypes and 71 mobile genetic element (MGE) subtypes were annotated across all samples. ARG Shannon diversity differed overall across groups (Kruskal-Wallis, p = 0.03); Bonferroni-adjusted Dunn’s test showed no significant pairwise differences. PCoA (Bray–Curtis) demonstrated distinct separation of the ST group (p = 0.002), and shared core ARG subtypes gradually increased with extended breeding years. A strong positive correlation between ARG and MGE abundances was identified (r = 0.85, p = 0.0001). In total, 63 contigs carrying co-localized ARG-MGE complexes were recovered. The ST group contained the highest proportion of such contigs. The ST group displayed tight physical ARG-MGE linkage within 1–3 kb genomic intervals. Conclusions: Our results reveal that breeding duration is associated with the gut ARG characteristics of captive forest musk deer. Short-term captivity has higher ARG-MGE co-localization, suggesting a higher possibility of mobilization.

Hemorrhagic pneumonia is a severe and often fatal disease in captive forest musk deer (Moschus berezovskii), but the pathogen remains incompletely understood. Based on incomplete statistics, the estimated incidence in captive populations ranges from 20% to 80%, with the disease occurring mainly in autumn, winter, and early spring. The disease has an acute onset and rapid progression. Due to the species’ strong stress response, affected animals rarely show behavioral changes, making early detection difficult. In this study, we investigated a mortality case presenting with oral bleeding and hematemesis on a forest musk deer farm. Postmortem examination revealed diffuse hemorrhagic pneumonia, and lung tissue samples were collected for histopathology, bacterial isolation, full-length 16S rRNA gene sequencing, and DNA/RNA virome sequencing. Histological examination showed extensive alveolar hemorrhage, fibrinous exudate, and macrophage infiltration. Bacterial culture and 16S rRNA gene sequencing identified Bergeyella zoohelcum as the predominant bacterium, accounting for 100% of the bacterial community in the lung tissue. Virome analysis revealed predominantly DNA bacteriophages (e.g., Cirlivirales, Cremevirales, Microviridae) and no known pathogenic RNA viruses; only seven low-abundance, unclassified RNA viral contigs of low completeness were detected. These results indicate that B. zoohelcum is the likely causative agent of hemorrhagic pneumonia in this case, with no evidence of viral involvement. This study provides the first direct association of B. zoohelcum with hemorrhagic pneumonia in forest musk deer, highlighting its pathogenic potential and the importance of monitoring this bacterium in captive populations.

Background: The nasopharyngeal microbiome is crucial for respiratory health in mammals, yet it remains poorly characterized in the endangered forest musk deer (Moschus berezovskii), particularly in the context of disease. Methods: We compared the bacterial (16S rRNA) and fungal (ITS2) communities in the nasopharynx of healthy (n = 6) and clinically diseased (n = 6) individuals. Results: Although alpha diversity did not differ significantly, beta diversity (PCoA) analysis revealed distinct bacterial (PERMANOVA, R² = 0.165, p = 0.014) and fungal (R² = 0.577, p = 0.003) community structures between groups. The diseased group exhibited a significant increase in the bacterial phylum Proteobacteria (70.97% vs. 46.27%), primarily driven by the genera Bibersteinia and Pseudomonas. Fungal communities in the diseased group were dominated by a higher relative abundance of Ascomycota and Basidiomycota, with significant enrichment of Wallemia and Aspergillus. LEfSe analysis identified Pseudomonas and multiple fungal taxa (e.g., Wallemia, Aspergillus) as biomarkers for the diseased group. PICRUSt2 prediction indicated enrichment of pathways related to carotenoid biosynthesis and sphingolipid metabolism in the diseased state, while FUNGuild analysis suggested a higher abundance of animal/plant pathogen-related fungi. Conclusions: Symptomatic respiratory infections in forest musk deer are associated with significant dysbiosis of the nasopharyngeal microbiome, characterized by the marked enrichment of potential bacterial opportunists (e.g., Pseudomonas) and specific fungal taxa (e.g., Wallemia, Aspergillus), alongside distinct functional shifts in the microbiome. These findings provide the first integrated bacterial–fungal profile of the nasopharyngeal microbiome in this endangered species, and highlight potential microbial biomarkers associated with respiratory disease.

Musk deer (Moschidae), a primitive lineage within Ruminantia, are distributed across East Asia and have long been of interest in molecular phylogenetic research. The spotted forest musk deer from Huanglong (HL) Mountain in Shaanxi, China, has long been morphologically classified as Moschus moschiferus (Siberian musk deer). However, its true taxonomic status has remained uncertain due to the lack of comprehensive molecular evidence. Moreover, studies on mitochondrial genome (mitogenome) variation within Moschidae, particularly at the intraspecific level, remain limited. To date, few phylogenetic analyses of Moschidae have incorporated all available complete mitochondrial genomes from public databases. In this study, we sequenced and assembled the complete mitogenomes of two spotted musk deer individuals from Huanglong Mountain, Shaanxi, China, and identified them as M. berezovskii (forest musk deer) by phylogenetic analysis. All available complete mitochondrial genomes of Moschidae were also included in the phylogenetic reconstruction. The average complete mitogenome length of M. berezovskii distributed in HL was 16,355 bp, and comprises 13 protein-coding genes (PCGs), two ribosomal RNA genes, and 22 transfer RNA genes. Phylogenetic analysis and genetic distance indicate that the taxonomic identity of the HL Mountain population as M. berezovskii supports the monophyly of M. berezovskii and provides a robust phylogenetic framework for clarifying evolutionary relationships within the family.

The relict gull (Larus relictus, Charadriiformes, Laridae) classified as vulnerable in the IUCN Red List is defined as a first-class national protected bird in China. However, our knowledge of the evolutionary history of L. relictus is limited. Here, we performed whole-genome resequencing of L. relictus (n = 14) and L. brunnicephalus (n = 3) to explore the genetic relationships and population structures and understand their adaptive evolution. The whole genome resequencing generated 667.55 Gb clean reads with an average sequencing depth of ~ 29×. The genomic variant analysis identified 13,717,267 heterozygous SNPs in the samples obtained from 17 individuals. Population genetic diversity analysis revealed that low nucleotide diversity (0.00029) and no obvious population structure in L. relictus. Demographic history revealed that from 180 to 5 kya (thousand years ago), the effective population size (Ne) of L. relictus exhibited declines (24,000 to 5,000), with a very low range population size (2,200 to 5,000). In contrast, from 100 to 80 kya, L. brunnicephalus peaked in ancestral Ne, followed by distinct declines at ~ 70 kya (100,000 to 16,000). The findings identified several genes associated with the correlated changed life-history traits of L. relictus, including BMP4 involved in beak adaptation; HAND2, NEUROG1, COL11A2, and EDNRB involved in the evolution of the palate, soft palate, and tongue; PIGR and PLCB2 involved in an enhanced response to bitter taste by sensing chemical secretions released by staple food substrate insects to activate protective mechanisms. Furthermore, thirty-four genes related to sperm development and activity, including KLHL10 and TEKT3, were identified in the expanded gene family. In addition, MED1, CNOT9, NR5A1, and PATZ1 were involved in enhanced male hormone secretion and a high density of candidate genes associated with embryonic development were identified. The findings indicated that the L. relictus population was in a male-biased diffusion mode; the function of the TEKT3 gene showed that males played a dominant role in brooding, which enhanced their attraction to females. Our study revealed that significant enrichment of olfactory signaling pathway genes, including OR14C36, OR14J1, OR14I1, and OR14A16; inner ear development-related, including PTN, PTPN11, GATA2, ATP8B1, and MYO15A; and those related to hypoxic adaptation to high-altitude breeding and iris colour. Based on the results and the knowledge of this organism biology and habitat use, we infer that less adaptive evolutionary pressure on vision in L. relictus were related with their feeding behaviour and adaptation. In summary, this comprehensive analysis provides insights into the evolutionary features of L. relictus and a new perspective for scientific research on L. relictus to effectively determine its future survival viability.

Background: Plovers (Charadriidae), within the order of Charadriiformes, a group of modern birds distributed worldwide, are a frequent subject of molecular phylogenetic studies. While research on mitochondrial genome (mitogenome) variation within the family Charadriidae, especially intraspecific variation, is limited. Additionally, the monophyly of Charadrius and the phylogenetic placement of Pluvialis remain contentious. Nevertheless, recent studies utilizing complete mitogenomes from available databases to construct phylogenetic trees for Charadriidae and Charadriiformes remain scarce. Methods: This study aims to explore mitogenome variation within Charadrius dubius and clarify the phylogenetic placement of Pluvialis fulva. We sequenced the complete mitogenome of six C. dubius and one P. fulva, and all additional available mitogenomes were integrated within Charadriiformes. The average complete mitogenome length of C. dubius is 16,889 bp, and P. fulva is 16,859 bp. Results: Our results support the suggestion that the monophyly of Charadrius and P. fulva is nested within Charadriidae. The phylogenetic analysis of Charadriiformes based on mitogenomes strongly supports the recognition of three major shorebird clades: Charadrii, Lari and Scolopaci, with Lari and Scolopaci identified as sister clades. Conclusions: Our study reinforces the credibility of the inferred evolutionary relationships within Charadriidae and Charadriiformes.

Breeding captive forest musk deer is an important way to obtain musk resources, but it also causes changes in the ecological environment of the breeding site. This study compared the changes in soil bacterial communities inside and outside the captive breeding site and tried to find out its influencing factors. High-throughput sequencing analysis and other methods were used to analyze the changes in the bacterial community structure in the natural soil and forest surrounding the musk deer site after 4 years of captivity. The results showed that the main dominant phyla in the captive soil samples were Proteobacteria, Acidobacteria and Chloroflexi. In order, Sphingomonadales (8.7%), Acidobacteriales (9.9%) and Solibacterales (6.1%) were dominant in the captive soil, while Rhizobiales (11.3%) and Gaiellales (6.2%) were found in non-captive soil. The main soil-specific microorganisms under captive conditions were Burkholderiales, Pseudomonadales and Sphingomadales. These microorganisms mainly gather at the order level and can be used as indicator microorganisms. A canonical correspondence analysis (CCA) showed that the microbial diversity in captivity soil was significantly affected by the contents of organic matter, available phosphorus and total nitrogen. These results will provide a basis for the healthy breeding of animals and shed light on the protection of the ecological environment in the Bashan Mountains of Qinling.

Hongjian Nur is an important breeding and stopover area for the globally endangered Relict gull (Larus relictus). This is where the species was first found in 2000. The breeding population of this species was monitored over the long term by directly counting nests from 2001 to 2020 in Hongjian Nur, Shaanxi, China. Our results suggest that breeding pairs increased rapidly, from 87 nests in 2001 to 7708 nests in 2010; at this point, the breeding population accounted for more than 85% of the global total, and was at the highest value during the past two decades. Subsequently, breeding pairs continued to decrease dramatically and reached a minimum number of 2054 nests in 2015, approximately 70% less than at their peak. In view of this situation, breeding islets were restored in 2014 and 2017, and the breeding population recovered slowly. Due to the changing ecological environment, breeding islets showed the same instability as the breeding population. Our conclusions support previous research, highlighting the importance of water level.

The mitochondrial genome (mitogenome) of Gallinago gallinago gallinago Linnaeus, 1758 was determined by the high-throughput data. The assembled mitogenome was 16,919 bp in length, with a 58.7% A + T content and GC skew of −0.3850. Among 13 PCGs, an unusual start codon (GTG) was identified for the COX1 gene, and incomplete stop codons (T-) were found in the COX3, ND2 and ND4 genes. The function of a cytosine insertion at site 174 in the ND3 gene and its phylogenetic significance are worthy of further scrutiny. In the control region (CR), thirteen 15-bp simple sequence repeats were found in G. g. gallinago. Phylogenetic analysis indicated that Gallinago was clustered at the basal position of the Scolopax clade and that the monophyly of Gallinago was also recovered. The mitogenome data of G. g. gallinago provides useful resources for further studying the evolution of Scolopacidae.

The complete mitogenome of the Lesser Whitethroat, Sylvia curruca blythi Ticehurst & Whistler, 1933 was determined, which belongs to Sylviidae, Passeriformes. The mitogenome had a length of 17,959 bp and consisted of 37 genes including 13 PCGs, 2 ribosomal RNAs (rRNA) genes, and 22 transfer RNAs (tRNA) genes. In addition, two control regions (CRs) were also existed in the mitogenome, with Sylvioidea typcial gene arrangement of cytb-trnT-CR1-trnP-nad6-trnE-CR2-trnF-rrnS. Phylogenetic analysis using 37 mitochondrial genes of 17 related species revealed that S. c. blythi had a closer relationship with S. crassirostris, and the monophyly of Sylvia was also recovered. The mitogenome data of S. c. blythi would provide useful resources for further studying the evolution of Sylvia and the subspecies taxonomic revision of S. curruca intraspecific.