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Abstract Immune evasion of Mycobacterium tuberculosis (Mtb) facilitates intracellular bacterial growth. The mechanisms of immune evasion, however, are still not fully understood. In this study, we reveal that tristetraprolin (TTP), one of the best characterized RNA‐binding proteins controlling the stability of targeted mRNAs, mediates innate immune evasion of mycobacteria. We found that TTP knockout mice displayed reduced bacterial burden in the early stage after Mtb aerosol challenge. Macrophages deficient in TTP also showed an inhibition in intracellular mycobacterial growth. Live mycobacteria induced TTP protein expression in macrophages, which was blocked by the mTOR inhibitor rapamycin. Rapamycin and AZD8055 specifically blocked 4EBP1 phosphorylation in infected macrophages and suppressed intracellular BCG growth. Rapamycin promoted TTP protein degradation through the ubiquitination pathway, whereas the proteasome inhibitor MG‐132 blocked rapamycin function and thus stabilized TTP protein. TTP induction suppressed the expression of iNOS/TNF‐α/IL‐12/IL‐23, and weakened protective immune responses in macrophages, whereas rapamycin enhanced the bactericidal effects through TTP inhibition. Moreover, blocking TTP binding increased the expression of TNF‐α and iNOS and suppressed intracellular mycobacterial growth. Overall, our study reveals a novel role for RNA‐binding protein TTP in Mtb immune evasion mechanisms and provides a potential target for host‐directed therapy against tuberculosis (TB).

Background The reproductive ability of male giant pandas has been a major complicating factor in the ex-situ conservation of the species. While it is well known that the testis produces sperm and secretes androgens, a process that requires precise regulation of various proteins, at present, there has been no systematic study on the composition of proteins in the testis of the giant pandas. Therefore, this study aims to apply proteomics to explore the regulation of proteins in the testes of giant pandas. Methods Samples from the testes of three giant pandas (22 years, 18 years, 8 days) were studied to assess the protein’s function. A label-free quantitative method was used to isolate testicular proteins from each male, 139,039 peptides and 11,435 proteins were obtained. Results Gene Ontology (GO) annotates most of the proteins involved in the processes of protein phosphorylation, oxidation-reduction, proteolysis, and signal transduction. KEGG pathway indicated that most of the proteins were involved in the pathway of signal transduction, transport, and catabolism. The protein kinase and WD40 repeats were involved in protein-protein interaction, which in turn regulates gene expression in the testicular tissue of giant pandas. Conclusions This study is the first to conduct an in-depth proteomic analysis of testicular tissue in giant pandas. The results revealed the important role of proteins in testicular tissue on spermatogenesis, testosterone production, and testicular microenvironment, providing clues for further research on male giant panda reproduction.

Establishing protected areas is a key element of most conservation strategies, but continuous effective management is needed to ensure that biodiversity is protected. Here we provide an evaluation of park effectiveness by quantifying changes in the abundance of the commonly occurring ungulates (bushbuck - Tragelaphus scriptus; red duiker - Cephalophus harveyi; blue duiker - Cephalophus moniticola, bushpigs - Potomochoerus larvatus, and giant forest hogs - Hylochoerus meinertzhageni), and African elephants (forest elephants - Loxodonta cyclotis, savanna elephants - Loxodonta africana, and their hybrids) in Kibale National Park, Uganda and consider the financial commitment needed to ensure protection. Following the upgrading of the protection of Kibale from a forest reserve to a national park, wildlife populations generally increased. Bushpigs are an exception and their populations appear stable. The increases of ungulate and elephant populations typically happened soon after park establishment, with the exception of the giant forest hog that only recently showed an increase. We conducted an assessment of Uganda Wildlife Authorities budget and those of the small NGOs working in the area and considered the contribution that Makerere University Biological Field Stations makes to the community. This analysis indicates that with a small budget UWA and its collaborators have been successful in protecting the park’s ungulate and elephant populations. While the past decades have demonstrated the park’s ability to conserve biodiversity with limited resources, the changing climate and mounting pressures faced by the park will necessitate significantly increased investments.

The giant panda (Ailuropoda melanoleuca), a vulnerable species and an icon of wildlife conservation, is still at risk due to habitat fragmentation and a low reproductive rate. To further safeguard the giant panda from extinction, a captive breeding program was established in the mid 1980ʼs, however the growth of this population has been hindered by the poor reproductive ability of captive male giant pandas. To address this, we investigated the sperm proteome of the giant panda as detailed information on sperm proteome is unavailable, as it is a highly specialized area of study. A study of adult panda sperm proteome identified 1921 proteins with enriched domains, including EF-hand, AAA+ ATPase, and WD 40 repeat. A comparison with four other species revealed common sperm proteins related to metabolic processes, especially glycolysis and citrate cycle, which are crucial for sperm energy. Panda-specific proteins were mainly associated with cellular protein metabolism. Serpin domain-related proteins may play a key role in panda semen properties and liquefaction, involving prefoldin beta-like and heat shock chaperonin-binding. This research contributes to understanding giant panda reproduction and aids in conservation efforts.

ABSTRACT Some lactic acid bacteria (LAB) can provide significant health benefits, which are critically important for the conservation of endangered animals, such as giant pandas. However, little is known about the diversity and culturability of LAB in the giant panda gut microbiota. To understand the roles of LAB in giant panda conservation, it is critical to culture bacterial strains of interest. In this study, we established a pipeline to culture bacterial strains using enrichment of target bacteria with different liquid media and growth conditions. Then, the strains were isolated in solid media to study their functions. Using 210 samples from the culture enrichment method and 138 culture-independent samples, we obtained 1120 amplicon sequencing variants (ASVs) belonging to Lactobacillales. Out of the 1120 ASVs, 812 ASVs from the culture enrichment approach were twofold more diverse than 336 ASVs from the culture-independent approach. Many ASVs of interest were not detected in the culture-independent approach. Using this pipeline, we isolated many relevant bacterial strains and established a giant panda gut bacteria strain collection that included strains with low-abundance in culture-independent samples and included most of the giant panda LAB described by other researchers. The strain collection consisted of 60 strains representing 35 species of 12 genera. Thus, our pipeline is powerful and provides guidance in culturing gut microbiota of interest in hosts such as the giant panda.IMPORTANCECultivation is necessary to screen strains to experimentally investigate microbial traits, and to confirm the activities of novel genes through functional characterization studies. In the long-term, such work can aid in the identification of potential health benefits conferred by bacteria and this could aid in the identification of bacterial candidate strains that can be applied as probiotics. In this study, we developed a pipeline with low-cost and user-friendly culture enrichment to reveal the diversity of LAB in giant pandas. We compared the difference between culture-independent and culture enrichment methods, screened strains of interest that produced high concentrations of short-chain fatty acids (SCFAs), and we investigated the catalog of virulence factors, antibiotic resistance, butyrate and lactate synthesis genes of the strains at a genomic level. This study will provide guidance for microbiota cultivation and a foundation for future research aiming to understand the functions of specific strains.

Objectives: To determine whether contrast-enhanced CT radiomics features can preoperatively predict lymphovascular invasion (LVI) and perineural invasion (PNI) in gastric cancer (GC). Methods: A total of 148 patients were included in the LVI group, and 143 patients were included in the PNI group. Three predictive models were constructed, including clinical, radiomics, and combined models. A nomogram was developed with clinical risk factors to predict LVI and PNI status. The predictive performance of the three models was mainly evaluated using the mean area under the curve (AUC). The performance of three predictive models was assessed concerning calibration and clinical usefulness. Results: In the LVI group, the predictive power of the combined model (AUC=0.871, 0.822) outperformed the clinical model (AUC=0.792, 0.728) and the radiomics model (AUC=0.792, 0.728) in both the training and testing cohorts. In the PNI group, the combined model (AUC=0.834, 0.828) also had better predictive power than the clinical model (AUC=0.764, 0.632) and the radiomics model (AUC=0.764, 0.632) in both the training and testing cohorts. The combined models also showed good calibration and clinical usefulness for LVI and PNI prediction. Conclusion: CECT-based radiomics analysis might serve as a non-invasive method to predict LVI and PNI status in GC.

Abstract Background Behaviors in captive animals, including changes in appetite, activity level, and social interaction, are often seen as adaptive responses. However, these behaviors may become progressively maladaptive, leading to stress, anxiety, depression, and other negative reactions in animals. Results In this study, we investigated the whole-genome sequencing data of 39 giant panda individuals, including 11 in captivity and 28 in the wild. To eliminate the mountain range effect and focus on the factor of captivity only, we first performed a principal component analysis. We then enumerated the 21,474,180 combinations of wild giant pandas (11 chosen from 28) and calculated their distances from the 11 captive individuals. The 11 wild individuals with the closest distances were used for the subsequent analysis. The linkage disequilibrium (LD) patterns demonstrated that the population was almost eliminated. We identified 505 robust selected genomic regions harboring at least one SNP, and the absolute frequency difference was greater than 0.6 between the two populations. GO analysis revealed that genes in these regions were mainly involved in nerve-related pathways. Furthermore, we identified 22 GO terms for which the selection strength significantly differed between the two populations, and there were 10 nerve-related pathways among them. Genes in the differentially abundant regions were involved in nerve-related pathways, indicating that giant pandas in captivity underwent minor genomic selection. Additionally, we investigated the relationship between genetic variation and chromatin conformation structures. We found that nucleotide diversity (θπ) in the captive population was correlated with chromatin conformation structures, which included A/B compartments, topologically associated domains (TADs) and TAD-cliques. For each GO term, we then compared the expression level of genes regulated by the above four factors (AB index, TAD intactness, TAD clique and PEI) with the corresponding genomic background. The retained 10 GO terms were all coordinately regulated by the four factors, and three of them were associated with nerve-related pathways. Conclusions This study revealed that giant pandas in captivity undergo short-term adaptation in nerve-related pathways. Furthermore, it provides new insights into the molecular mechanism of gene expression regulation under short-term adaptation to environmental change.

The evolutionary development and phylogenetic division between Asian and African cercopithecoids (Cercopithecidae) have attracted significant attention in genetics, molecular biology, behavior, and morphology. However, less emphasis has been placed on how they have evolved morphologically after divergence, approximately 10 million years ago (mya) for Colobinae and 5–7 mya for Cercopithecinae, corresponding to the significant variation and diversity in landscape, climate, habitat, and ecologies between the two continents. This study examines whether such variation and diversity have been reflected in dental morphology. Our findings reveal substantial differences between Hylobatidae and Cercopithecidae, as well as between Colobinae and Cercopithecinae, indicating that size-adjusted dental variation mainly reveals the diversity associated with evolution and phylogenetic inertia. Interestingly, despite the earlier divergence of Afro-Asian colobines, their Euclidean Distance is comparable to that of Afro-Asian cercopithecines. This implies that latecomers (macaques) demonstrate equivalent diversity to colobines due to their extensive dispersion and broader adaptative radiation on the same continent. Colobinae exhibit more developed premolar and molar regions. However, when post-canine teeth are considered alone, Colobinae present a significantly larger molar size than Asian Cercopithecinae but not with the African Cercopihecinae. This contradicts the hypothesis that folivorous primates (Colobinae) have larger post-canine molars than frugivorous ones (Cercopithecinae). The considerable molar size in African Cercopithecinae must be associated with their more protrusive and larger facial structure rather than a specific dietary preference, being less diverse than their Asian counterparts—a trait that has evolved phylogenetically. This study also paves the way for further exploration of facial and cranial differences between the continental groups of Cercopithecinae and Colobinae, delving deeply into diversity variation due to geographical and climatic adaptations.

Wild giant pandas are inherently solitary creatures, however, the ex-situ conservation efforts significantly alter the living circumstances of their captive counterparts. Following the breeding period, giant pandas in captivity may be maintained in social groups. Currently, there is a lack of research on the effects of group housing on the physiology, behavior, and gut microbiota of captive giant pandas. This study divided six captive giant pandas into two groups following the breeding period. By comparing the behavior, physiology, and microorganisms of the two groups, we aim to investigate the behavioral responses and physiological adaptation mechanisms exhibited by captive giant pandas in a “group living” state. Our findings indicate that sub-adult giant pandas housed in group settings exhibit a significantly longer duration of playing behavior (including interactive and non-interactive play) compared to their counterparts housed separately (p < 0.001) while also demonstrating a significantly lower duration of stereotyped behavior than their separately housed counterparts. Additionally, an analysis of urine cortisol and heart rate variability between the two groups revealed no significant differences. Simultaneously, the group housing strategy markedly elevated the β diversity of gut microbiota in sub-adult giant pandas. In conclusion, the group-rearing model during the sub-adult stage has been shown to significantly alter the behavioral patterns of captive giant pandas. In conclusion, within the present captive setting, the group-rearing approach during the sub-adult stage proved to be less distressing for adult captive giant pandas.

Abstract Fallback foods (FBF), categorized into staple and filler types, are suboptimal food sources chosen by animals in response to a scarcity of preferred food items during specific periods. Using lichens as FBF by Yunnan snub‐nosed monkeys (Rhinopithecus bieti) represents a distinctive ecological adaptation and evolutionary development within nonhuman primates. This study delves into the annual dietary choices of the species to address issues, elucidate the nutritional value, and understand the ecological significance of lichens for this primate species, which resides at the highest altitudes and experiences the coldest weather among global primates. The findings reveal that the lichens consumed by the monkeys serve as the staple FBF, with Bryoria spp. and Usnea longissima being the primary dietary species. The former is the preferred choice, providing higher digestible fiber (neutral detergent fiber) levels but lower tannin, fat, ADF, and energy levels. During the dry season, lichens dominate as the monkeys' primary food and nutritional resources. In the wet season, they act as a fundamental food selection rather than an ideal dietary choice, substituting nutrients from fruits, seeds, and leaves. Compared to other Asian colobine counterparts, this species exhibits the highest lichen consumption but the lowest proportions of leaves, flowers, and seeds. This study provides valuable evidence and information for developing or amending conservation strategies and guidelines for the dietary management of captive breeding of monkeys, one of the world's critically endangered primate species.

Red pandas evolved from carnivores to herbivores and are unique within Carnivora. Red pandas and carnivorous mammals consume milk during the suckling period, while they consume bamboo and meat during the adult period, respectively. Red pandas and carnivorous mammal ferrets have a close phylogenetic relationship. To further investigate the molecular mechanisms of dietary changes and nutrient utilization in red pandas from suckling to adult, comparative analysis of the whole transcriptome was performed on stomach tissues from red pandas and ferrets during the suckling and adult periods. The main results are as follows: (1) we identified ncRNAs for the first time in stomach tissues of both species, and found significant expression changes of 109 lncRNAs and 106 miRNAs in red pandas and 756 lncRNAs and 109 miRNAs in ferrets between the two periods; (2) up-regulated genes related to amino acid transport regulated by lncRNA-miRNA-mRNA networks may efficiently utilize limited bamboo amino acids in adult red pandas, while up-regulated genes related to amino acid degradation regulated by lncRNAs may maintain the balance of amino acid metabolism due to larger daily intakes in adult ferrets; and (3) some up-regulated genes related to lipid digestion may contribute to the utilization of rich nutrients in milk for the rapid growth and development of suckling red pandas, while up-regulated genes associated with linoleic acid metabolism regulated by lncRNA-miRNA-mRNA networks may promote cholesterol decomposition to reduce health risks for carnivorous adult ferrets. Collectively, our study offers evidence of gene expression adaptation and ncRNA regulation in response to specific dietary changes and nutrient utilization in red pandas during suckling and adult periods.

The goal of this study is to present an automatic vocalization recognition system of giant pandas (GPs). Over 12800 vocal samples of GPs were recorded at Chengdu Research Base of Giant Panda Breeding (CRBGPB) and labeled by CRBGPB animal husbandry staff. These vocal samples were divided into 16 categories, each with 800 samples. A novel deep neural network (DNN) named 3Fbank-GRU was proposed to automatically give labels to GP's vocalizations. Unlike existing human vocalization recognition frameworks based on Mel filter bank (Fbank) which used low-frequency features of voice only, we extracted the high, medium and low frequency features by Fbank and two self-deduced filter banks, named Medium Mel Filter bank (MFbank) and Reversed Mel Filter bank (RFbank). The three frequency features were sent into the 3Fbank-GRU to train and test. By training models using datasets labeled by CRBGPB animal husbandry staff and subsequent testing of trained models on recognizing tasks, the proposed method achieved recognition accuracy over 95%, which means that the automatic system can be used to accurately label large data sets of GP vocalizations collected by camera traps or other recording methods.

Ticks and tick-borne diseases have negative impacts on the health of wild animals including endangered and vulnerable species. The giant panda (Ailuropoda melanoleuca), a vulnerable and iconic flagship species, is threatened by tick infestation as well. Not only can ticks cause anemia and immunosuppression in the giant panda, but also bacterial and viral diseases. However, previous studies regarding tick infestation on giant pandas were limited in scope as case reports from sick or dead animals. In this study, an investigation focusing on the tick infestation of a reintroduced giant panda at the Daxiangling Reintroduction Base in Sichuan, China was conducted. Ticks were routinely collected and identified from the ears of the giant panda from March to September in 2021. A linear model was used to test the correlation between tick abundance and climate factors. All ticks were identified as Ixodes ovatus. Tick abundance was significantly different among months. Results from the linear model showed temperature positively correlated to tick abundance, while air pressure had a negative correlation with tick abundance. To the best of our knowledge, this study is the first reported investigation of tick species and abundance on a healthy giant panda living in the natural environment, and provides important information for the conservation of giant pandas and other species sharing the same habitat.

Abstract Background It is inevitable to change the function or expression of genes during the environmental adaption of species. Both the giant panda (Ailuropoda melanoleuca) and red panda (Ailurus fulgens) belong to Carnivora and have developed similar adaptations to the same dietary switch to bamboos at the morphological and genomic levels. However, the genetic adaptation at the gene expression level is unclear. Therefore, we aimed to examine the gene expression patterns of giant and red panda convergent specialized bamboo-diets. We examined differences in liver and pancreas transcriptomes between the two panda species and other non-herbivorous species. Results The clustering and PCA plots suggested that the specialized bamboo diet may drive similar expression shifts in these two species of pandas. Therefore, we focused on shared liver and pancreas DEGs (differentially expressed genes) in the giant and red panda relative to other non-herbivorous species. Genetic convergence occurred at multiple levels spanning carbohydrate metabolism, lipid metabolism, and lysine degradation. The shared adaptive convergence DEGs in both organs probably be an evolutionary response to the high carbohydrate, low lipid and lysine bamboo diet. Convergent expression of those nutrient metabolism-related genes in both pandas was an intricate process and subjected to multi-level regulation, including DNA methylation and transcription factor. A large number of lysine degradation and lipid metabolism related genes were hypermethylated in promoter regions in the red panda. Most genes related to carbohydrate metabolism had reduced DNA methylation with increased mRNA expression in giant pandas. Unlike the red panda, the core gene of the lysine degradation pathway (AASS) doesn’t exhibit hypermethylation modification in the giant panda, and dual-luciferase reporter assay showed that transcription factor, NR3C1, functions as a transcriptional activator in AASS transcription through the binding to AASS promoter region. Conclusions Our results revealed the adaptive expressions and regulations of the metabolism-related genes responding to the unique nutrients in bamboo food and provided data accumulation and research hints for the future revelation of complex mechanism of two pandas underlying convergent adaptation to a specialized bamboo diet.

As a typical solitary animal, adult giant pandas rely on chemical signals (sex pheromones) to transmit reproductive information during oestrous. Although researchers have confirmed that the gut microbiota is related to the emission and reception of sex pheromones, there is no clear correlation between the gut microbes and the synthesis of sex pheromone of giant pandas, that is, which gut microbes and microbial metabolites are participate in the synthesis of giant panda’s sex pheromone. As a mirror of gut microbiota, fecal microbiota can reflect the composition of gut microbiota and its interaction with host to some extent. The purpose of this study is to explore how the gut microbes affect the synthesis of sex pheromones in captive giant pandas by combining analysis of the fecal microbiome and metabolomics. The results of correlation and microbial function analysis show that intestinal microorganisms such as Veillonellaceae and Lactobacillilaceae are associated with the synthesis of short chain fatty acid (acetic acid) and volatile ester metabolites, such as 1-butanol, 3-methyl, acetate, acetic acid, hexyl ester and 3-hexen-1-ol, acetate, (Z). In summary, based on this study, we believe that volatile metabolites such as fecal acetate participate in the process of mate preference of captive giant pandas and affect their expression of natural mating behavior. The possible mechanism is that the gut microbes can promote the synthesis of key chemical signaling substances in perianal glands through mediated intermediate fecal metabolites, thus affecting the normal information exchange between giant pandas individuals. The results of this study have greatly enriched our understanding of gut microbes regulating the synthesis of sex pheromones in giant pandas.

ABSTRACT The gut flora is a treasure house of diverse bacteriophages maintaining a harmonious and coexistent relationship with their hosts. The giant panda (Ailuropoda melanoleuca), as a vulnerable endemic species in China, has existed for millions of years and is regarded as a flagship species for biodiversity conservation. And yet, limited studies have analyzed the phage communities in the gut of giant pandas. Using viral metagenomic analysis, the phageomes of giant pandas and other relative species were investigated. Our study explored and compared the composition of phage communities from different animal sources. Giant pandas possessed more diverse and abundant phage communities in the gut compared with other relevant animals. Phylogenetic analyses based on the phage terminase large subunit (TerL) showed that the Caudovirales phages in giant pandas also presented highly genetic diversity. Our study revealed the diversity of phage communities in giant pandas and other relative species, contributing to the health maintenance of giant pandas and laying the groundwork for molecular evolution research of bacteriophages in mammals. IMPORTANCE Gut phageome plays an important role in shaping gut microbiomes by direct interactions with bacteria or indirect influences on the host immune system, potentially regulating host health and disease status. The giant panda (Ailuropoda melanoleuca) is a vulnerable and umbrella species for biodiversity conservation. Our work explored and compared the gut phageome of giant pandas and relative species, contributing to the health maintenance of giant pandas.

There are abundant resources and many endangered marine animals in the ocean. Using sound to effectively identify and locate them, and estimate their distribution area, has a very important role in the study of the complex diversity of marine animals (Hanny et al., 2013). We design a Two-Stream ConvNet with Attention (TSCA) model, which is a two-stream model combined with attention, in which one branch processes the temporal signal and the other branch processes the frequency domain signal; It makes good use of the characteristics of high time resolution of time domain signal and high recognition rate of frequency domain signal features of sound, and it realizes rapid localization and recognition of sound of marine species. The basic network architecture of the model is YOLO (You Only Look Once) (Joseph et al., 2016). A new loss function focal loss is constructed to strengthen the impact on the tail class of the sample, overcome the problem of data imbalance and avoid over fitting. At the same time, the attention module is constructed to focus on more detailed sound features, so as to improve the noise resistance of the model and achieve high-precision marine species identification and location. In The Watkins Marine Mammal Sound Database, the recognition rate of the algorithm reached 92.04% and the positioning accuracy reached 78.4%.The experimental results show that the algorithm has good robustness, high recognition accuracy and positioning accuracy.

In the process of industrial production, manual assembly of workpieces exists with low efficiency and high intensity, and some of the assembly process of the human body has a certain degree of danger. At the same time, traditional machine learning algorithms are difficult to adapt to the complexity of the current industrial field environment; the change in the environment will greatly affect the accuracy of the robot’s work. Therefore, this paper proposes a method based on the combination of machine vision and the YOLOv5 deep learning model to obtain the disk porous localization information, after coordinate mapping by the ROS communication control robotic arm work, in order to improve the anti-interference ability of the environment and work efficiency but also reduce the danger to the human body. The system utilizes a camera to collect real-time images of targets in complex environments and, then, trains and processes them for recognition such that coordinate localization information can be obtained. This information is converted into coordinates under the robot coordinate system through hand–eye calibration, and the robot is then controlled to complete multi-hole localization and tracking by means of communication between the upper and lower computers. The results show that there is a high accuracy in the training and testing of the target object, and the control accuracy of the robotic arm is also relatively high. The method has strong anti-interference to the complex environment of industry and exhibits a certain feasibility and effectiveness. It lays a foundation for achieving the automated installation of docking disk workpieces in industrial production and also provides a more favorable choice for the production and installation of the process of screw positioning needs.

Toxoplasma gondii is a worldwide-distributed zoonotic protozoan parasite which causes toxoplasmosis and has a significant effect on public health. In the giant panda (Ailuropoda melanoleuca), toxoplasmosis can cause asymptomatic infections, reproductive disorder and even death, which poses a serious threat to the conservation of this rare protected species. Therefore, serological investigation of T. gondii is essential to understanding its risk to giant pandas, however, there are no specific testing kits for giant pandas. Previous research has used MAT as the reference method for screening T. gondii, to investigate this further, this study focused on the agreement comparing of MAT with ELISA and IHA tests for detecting T. gondii antibodies in 100 blood samples from 55 captive giant pandas in Chengdu, China. The results showed 87.0%, 87.0%, 84.0%, samples were sero-positive for T. gondii using ELISA (kits a, b, c), respectively, while MAT and IHA tests were 84.0% and 9.0% sero-positive, respectively. There was no significant difference between MAT and the three ELISA kits and these two methods had substantial agreement (0.61

Abstract Background Extended-spectrum β-lactamases (ESBL)-producing strains of Klebsiella pneumoniae remain a worldwide, critical clinical concern. However, limited information was available concerning ESBL-producing Klebsiella pneumoniae in giant pandas. The objective of this study was to characterize ESBL-producing Klebsiella pneumoniae isolates from captive giant pandas. A total of 211 Klebsiella pneumoniae isolates were collected from 108 giant pandas housed at the Chengdu Research Base of Giant Panda Breeding (CRBGP), China. Samples were screened for the ESBL-producing phenotype via the double-disk synergy test. Result A total of three (1.42%, n = 3/211) ESBL-producing Klebsiella pneumoniae strains were identified, and characterization of ESBL-producing Klebsiella pneumoniae isolates were studied by the detection of ESBL genes and mobile genetic elements (MGEs), evaluation of antimicrobial susceptibility and detection of associated resistance genes. Clonal analysis was performed by multi-locus sequencing type (MLST). Among the three ESBL-producing isolates, different ESBL-encoding genes, including bla CTX-M, and bla TEM, were detected. These three isolates were found to carry MGEs genes (i.e., IS903 and tnpU) and antimicrobial resistance genes (i.e., aac(6')-Ib, aac(6')-I, qnrA, and qnrB). Furthermore, it was found that the three isolates were not hypermucoviscosity, resistant to at least 13 antibiotics and belonged to different ST types (ST37, ST290, and ST2640). Conclusion Effective surveillance and strict infection control strategies should be implemented to prevent outbreaks of ESBL-producing Klebsiella pneumoniae in giant pandas.

The spread of antibiotic resistance genes (ARGs) in the environment exacerbates the contamination of these genes; therefore, the role plants play in the transmission of resistance genes in the food chain requires further research. Giant pandas consume different bamboo parts at different times, which provides the possibility of investigating how a single food source can affect the variation in the spread of ARGs. In this study, metagenomic analysis and the Comprehensive Antibiotic Resistance Database (CARD) database were used to annotate ARGs and the differences in gut microbiota ARGs during the consumption of bamboo shoots, leaves, and culms by captive giant pandas. These ARGs were then compared to investigate the impact of bamboo part consumption on the spread of ARGs. The results showed that the number of ARGs in the gut microbiota of the subjects was highest during the consumption of bamboo leaves, while the variety of ARGs was highest during the consumption of shoots. Escherichia coli, which poses a higher risk of ARG dissemination, was significantly higher in the leaf group, while Klebsiella, Enterobacter, and Raoultella were significantly higher in the shoot group. The ARG risk brought by bamboo shoots and leaves may originate from soil and environmental pollution. It is recommended to handle the feces of giant pandas properly and regularly monitor the antimicrobial and virulence genes in their gut microbiota to mitigate the threat of antibiotic resistance.

Abstract Background The red panda has been classified as an endangered species due to the decreased number in the world and disease is considered as a great threat to the health and survival of the cubs in captivity. Results This study analyzed 32 red panda cub mortalities (15 females and 17 males, age less than two months) through gross necropsy, microbiological examination, and histopathological observation at the Chengdu Research Base of Giant Panda Breeding, China, during 2014–2020. The results showed that screenings for canine distemper virus, canine parvovirus, rotavirus and parasite infection were all negative, however bacteria such as Klebsiella pneumoniae, Proteus mirabilis, Escherichia coli, Enterococcus faecalis, Pseudomonas were isolated from the tissue samples of some cubs. The major causes of death were respiratory (43.75%) and digestive system disease (28.13%), followed by cardiovascular disease (12.5%) and neonatal stillbirths (9.38%). Renal system diseases and trauma were also detected, at lower incidence (one case for each). The mortality rate within 15 days of birth was 68.75% and gradually decreased with age, there was no significant difference in gender. Conclusion This study can provide a scientific basis for the analysis of the cause of death among red panda cubs in captivity, so as to improve the survival rate, help build the captive population and further the ex-situ conservation management of this endangered species. Additionally, our research may also provide insights into the in-situ conservation of wild red pandas by identifying emerging disease threats within the wild population and potential treatment for rescued individuals.

Summary: Tactical deception can be beneficial for social animals during intra-specific competition. However, the use of tactical deception in wild mammals is predicted to be rare. We tested whether a food-provisioned free-ranging band of golden snub-nosed monkeys (Rhinopithecus roxellana) use alarm calls in a functionally deceptive manner to gain access to food resources, whether the rate of deceptive alarm calls varies among individuals, and whether there are any counter-deception behaviors. We used a hexagonal camera array consisting of 10 cameras to record videos during feeding, which allowed us to identify individual alarm callers. We found evidence that these monkeys use deceptive alarms and that adult females were more likely to use such calls than other individuals. The monkeys increased their rates of response to alarm calls when competition for food was high. However, we found no direct evidence of any counter-deception strategies.

Mature female giant pandas usually ovulate once a year. This is followed by an obligatory luteal phase, consisting of a long-lasting corpus luteum dormancy phase (CLD; primary increase in progestogens) and a much shorter active luteal phase (AL; secondary increase in progestogens). Varying duration of both the dormant (embryonic diapause) and AL (post-embryo reactivation) phases has hampered unambiguous pregnancy length determination in giant pandas until today. Additionally, progestogen profiles have been considered not to differ between pregnant and pseudopregnant cycles. Only ceruloplasmin, 13,14-dihydro-15-keto-PGF2α (PGFM) and – more recently – estrogens have been assigned diagnostic power so far. Our study investigated the competence of metabolic (fecal output) and Urinary Specific Gravity (USpG)-normalized urinary endocrine (progestogens, PGFM, glucocorticoids (GCM) and ceruloplasmin) markers for pregnancy monitoring including defining the duration of the AL phase length. Research on 24 (6 pregnant, 8 pseudopregnant and 10 non-birth) cycles of 6 giant pandas revealed a fixed AL phase length of 42 days in giant pandas, e.g. representing 6 weeks of post- diapause development in case of pregnancy. Progestogen concentrations were significantly higher in pregnant cycles throughout the majority of the AL phase, with significant higher values during the AL phase in healthy twin compared to singleton pregnancies. GCM concentrations were also markedly higher in giant pandas expecting offspring, with a clear increase towards birth in the final 2 weeks of pregnancy. This increase in GCM was running in parallel with elevating estrogen and PGFM concentrations, and decreasing progestogens. In addition, during the AL phase, a more pronounced decrease in fecal output was obvious for pregnant females. The combined profiles of non-invasive metabolic and endocrine markers, the latter normalized based on USpG, showed a true pregnancy signature during the AL phase. The findings of this study are applicable to retrospective evaluations of non-birth cycles facilitating categorizing those into pseudopregnant or lost pregnancies, with USpG-normalization of the urinary endocrine markers as a prerequisite.

Hyperspectral images are a valuable tool for remotely sensing important characteristics of a variety of landscapes, including water quality and the status of marine disasters. However, hyperspectral data are rare or expensive to obtain, which has spurred interest in low-cost, fast methods for reconstructing hyperspectral data from much more common RGB images. We designed a novel algorithm to achieve this goal using multi-scale atrous convolution residual network (MACRN). The algorithm includes three parts: low-level feature extraction, high-level feature extraction, and feature transformation. The high-level feature extraction module is composed of cascading multi-scale atrous convolution residual blocks (ACRB). It stacks multiple modules to form a depth network for extracting high-level features from the RGB image used as an input. The algorithm uses jump connection for residual learning, and the final high-level feature combines the output of the low-level feature extraction module and the output of the cascaded atrous convolution residual block element by element, so as to prevent gradient dispersion and gradient explosion in the deep network. Without adding too many parameters, the model can extract multi-scale features under different receptive fields, make better use of the spatial information in RGB images, and enrich the contextual information. As a proof of concept, we ran an experiment using the algorithm to reconstruct hyperspectral Sentinel-2 satellite data from the northern coast of Australia. The algorithm achieves hyperspectral spectral reconstruction in 443nm-2190nm band with less computational cost, and the results are stable. On the Realworld dataset, the reconstruction error MARE index is less than 0.0645, and the reconstruction time is less than 9.24S. Therefore, in the near infrared band, MACRN reconstruction accuracy is significantly better than other spectral reconstruction algorithms. MACRN hyperspectral reconstruction algorithm has the characteristics of low reconstruction cost and high reconstruction accuracy, and its advantages in ocean spectral reconstruction are more obvious.

To contribute to the conservation of endangered animals, the utilization of model systems is critical to analyze the function of their gut microbiota. In this study, the results of a fecal microbial transplantation (FMT) experiment with germ-free (GF) mice receiving giant panda or horse fecal microbiota showed a clear clustering by donor microbial communities in GF mice, which was consistent with the results of blood metabolites from these mice. At the genus level, FMT re-established approximately 9% of the giant panda donor microbiota in GF mice compared to about 32% for the horse donor microbiota. In line with this, the difference between the panda donor microbiota and panda-mice microbiota on whole-community level was significantly larger than that between the horse donor microbiota and the horse-mice microbiota. These results were consistent with source tracking analysis that found a significantly higher retention rate of the horse donor microbiota (30.9%) than the giant panda donor microbiota (4.0%) in GF mice where the microbiota remained stable after FMT. Further analyzes indicated that the possible reason for the low retention rate of the panda donor microbiota in GF mice was a low relative abundance of Clostridiaceae in the panda donor microbiota. Our results indicate that the donor microbiota has a large effect on GF mice microbiota after FMT.

Abstract Knowledge of energy expenditure informs conservation managers for long term plans for endangered species health and habitat suitability. We measured field metabolic rate (FMR) of free-roaming giant pandas in large enclosures in a nature reserve using the doubly labeled water method. Giant pandas in zoo like enclosures had a similar FMR (14,182 kJ/day) to giant pandas in larger field enclosures (13,280 kJ/day). In winter, giant pandas raised their metabolic rates when living at − 2.4 °C (36,108 kJ/day) indicating that they were below their thermal neutral zone. The lower critical temperature for thermoregulation was about 8.0 °C and the upper critical temperature was about 28 °C. Giant panda FMRs were somewhat lower than active metabolic rates of sloth bears, lower than FMRs of grizzly bears and polar bears and 69 and 81% of predicted values based on a regression of FMR versus body mass of mammals. That is probably due to their lower levels of activity since other bears actively forage for food over a larger home range and pandas often sit in a patch of bamboo and eat bamboo for hours at a time. The low metabolic rates of giant pandas in summer, their inability to acquire fat stores to hibernate in winter, and their ability to raise their metabolic rate to thermoregulate in winter are energetic adaptations related to eating a diet composed almost exclusively of bamboo. Differences in FMR of giant pandas between our study and previous studies (one similar and one lower) appear to be due to differences in activity of the giant pandas in those studies.

Abstract Background Tropical forests are repositories of much of the world’s biodiversity and are critical for mitigation of climate change. Yet, the drivers of forest dynamics are poorly understood. This is in large part due to the lack of long-term data on forest change and changes in drivers. Methodology We quantify changes in tree abundance, diversity, and stand structure along transects first enumerated in 1978 and resampled 2019 in Kibale National Park, Uganda. We tested five predictions. First, based on the purported role of seed dispersal and herbivory and our quantification of changes in the abundance of frugivores and herbivores, we tested two predictions of how faunal change could have influenced forest composition. Second, based on an evaluation of life history strategies, we tested two predictions concerning how the forest could have changed following disturbance that happened prior to written history. Finally, based on a 50-year climate record, we evaluate the possible influence of climate change on forest dynamics. Results More trees were present on the assessed transects in 2019 (508) than in 1978 (436), species richness remained similar, but diversity declined as the number of dominant species increased. Rainfall increased by only 3 mm over the 50 years but this had not significant effect on forest changes measured here. Annual average monthly maximum temperature increased significantly by 1.05 °C over 50 years. The abundance of frugivorous and folivorous primates and elephants increased over the 50 years of monitoring. Neither the prediction that an increase in abundance of seed dispersing frugivores increases the abundance of their preferred fruiting tree species, nor that as an increase in folivore abundance causes a decline in their preferred species were supported. As predicted, light-demanding species decreased in abundance while shade-tolerant species increased as expected from Kibale being disturbed prior to historical records. Finally, while temperature increased over the 50 years, we found no means to predict a priori how individual species would respond. Conclusions Our study revealed subtle changes in the tree community over 40 years, sizable increases in primate numbers, a substantial increase in the elephant population and an increase in local temperature. Yet, a clear picture of what set of interactions impact the change in the tree community remains elusive. Our data on tree life-history strategies and frugivore/herbivore foraging preferences suggest that trees species are under opposing pressures.

Abstract Background Flavonoids are important plant secondary metabolites (PSMs) that have been widely used for their health-promoting effects. However, little is known about overall flavonoid metabolism and the interactive effects between flavonoids and the gut microbiota. The flavonoid-rich bamboo and the giant panda provide an ideal system to bridge this gap. Results Here, integrating metabolomic and metagenomic approaches, and in vitro culture experiment, we identified 97 flavonoids in bamboo and most of them have not been identified previously; the utilization of more than 70% flavonoid monomers was attributed to gut microbiota; the variation of flavonoid in bamboo leaves and shoots shaped the seasonal microbial fluctuation. The greater the flavonoid content in the diet was, the lower microbial diversity and virulence factor, but the more cellulose-degrading species. Conclusions Our study shows an unprecedented landscape of beneficial PSMs in a non-model mammal and reveals that PSMs remodel the gut microbiota conferring host adaptation to diet transition in an ecological context, providing a novel insight into host-microbe interaction. Video abstract

IntroductionToxoplasma gondii, a globally zoonotic protozoan parasite, infects most warm-blooded animals including the giant panda, and poses a serious threat to the giant panda conservation. However, the seroprevalence and the risk factors for toxoplasmosis in giant pandas are unknown. Here we aimed to determine the seroprevalence of T. gondii in the captive population of giant pandas and analyze the factors associated with the increased risk of infection.MethodsA total of 203 serum samples were collected from 157 (95 females and 62 males) captive giant pandas from 2007 to 2022, antibodies against T. gondii were screened using commercial ELISA and MAT kits.ResultsThe results showed 56 (35.67%) giant pandas were seropositive, age and transfer history between institutions were identifified as risk factors for T. gondii infection. It is suggested that age-related seroprevalence was the main factor, and housing multiple species in the same environment may increase the chance of cross-infection of T. gondii.DiscussionThis study can provide research data for developing policies for the prevention and control of T. gondii and protecting the health of captive giant pandas and other wildlife.

To achieve reproduction, male solitary mammals need to locate females using chemical communication with high levels of precision. In the case of giant pandas, the total estrus period of females was usually 15 days each year, however, successful mating activity is finished within 3 days from respective home range. The mating pattern of giant pandas, where multiple males compete for each female requires females employ efficient systems to communicate their estrus phases. To verifying whether the scent secretions of giant pandas changes by gender and estrus progression, the microbiota and compounds in 29 anogenital gland samples from 14 individuals during estrus were analyzed by 16S rRNA sequencing and GC-MS. We show that the microbiota communities covary by gender with 4 particular compounds of scent secretions. Among 597 genera, 34 were identified as biomarkers that could be used to distinguish between different estrus phases. By bacterial-compounds co-analysis, 3 fatty ester acids and squalene compounds covaried with the development of estrus in the bacterial communities of female giant pandas. This study helps clarify how a large, solitary mammal expresses accurate information to improve the likelihood of successful reproduction by changing the composition of microbiota and odor compounds of anogenital glands during estrus.

Summary: Evolutionary and historical development and current profiles are essential to generating a tangible conservation strategy. It is also critical to distinguish the regions with vigorous potential growth from those meeting evolutionary development bottlenecks and those whose development has been severely devastated. We used two sizeable national data repositories of terrestrial fauna and flora of China to approach the issues. The results indicate that the Southwest and Coastal regions have the most significant terrestrial faunal-floral biodiversity (TFFB). Thus, they should be prioritized in conservation for great potential promotions. Although there has been remarkable evolutionary development, the Central region has been severely devastated. A solution is to uphold a balanced association between social-economic development and TFFB sustainability. As for the Northeast and the western Northwest, there is no need to invest heavily in conservation measures. This study sheds light on exploring more practical conservation strategies regionally, nationally, and globally to achieve pragmatic goals.

The decline in natural mating behavior is the primary reason underlying in the poor population growth of captive giant pandas. However, the influencing factors and underlying mechanisms remain unclear to data. It is speculated that the decline in natural mating behavior could be related to the psychological stress caused by captivity, which restricts their free choice of mates. In order to test this hypothesis, we performed urinary metabolomics analysis using Ultra-High-Performance Liquid Chromatography-Mass Spectrometry (UHPLC/-MS) combined with 16S rDNA sequencing for exploring the physiological mechanism underlying the decline in the natural mating behavior of captive giant panda. The results demonstrated that the decline in mating ability could be related to abnormalities in arginine biosynthesis and neurotransmitter synthesis. Additionally, the relative abundance of bacteria from the Firmicutes, Proteobacteria, and Actinobacteria phyla and the Acinetobacter, Weissella, and Pseudomonas genus was significantly reduced in the group with low natural mating behavior. These findings imply that the inhibition of arginine synthesis induced by environmental changes could be related to the poor libido and failure of mate selection in captive giant pandas during the breeding period. The results also demonstrate the relationship between the altered urinary microbes and metabolites related to arginine and neurotransmitter synthesis. These findings may aid in understanding the mechanism underlying environment-induced mate selection in captive giant pandas and propose a novel strategy for determining the sexual desire of giant pandas based on urinary microbes. The method would be of great significance in improving the natural reproductive success rate of captive giant pandas.

The objective of this study was to understand biological characteristics of one bacteria strain named as VPG which was isolated from multiple organs of a dead captive giant panda cub. Here, we use biochemical tests, 16S rRNA and gyrB genes for bacterial identification, the disk diffusion method for antibiotic resistance phenotype, smart chip real-time PCR for the antibiotic resistance genotype, multiplex PCR for determination of virulence genes, and the acute toxicity test in mice for testing the pathogenicity of isolates. The isolate was identified as A. veronii strain based on the biochemical properties and genetic analysis. We found that the strain carried 31 antibiotic resistance genes, revealed antimicrobial resistance phenotypically to several antibiotics including penicillin, ampicillin, oxacillin, amoxicillin, imipenem, and vancomycin, and carried virulence genes including aer, act, lip, exu, ser, luxs, and tapA. The main pathological changes in giant panda were congestion, necrotic lesions and a large number of bacteria in multiple organs. In addition, the LD50 in Kunming mice infected with strain VGP was 5.14 × 107 CFU/mL by intraperitoneal injection. Infection with strain VGP led to considerable histological lesions such as hemorrhage of internal organs, necrosis of lymphocytes and neurons in Kunming mice. Taken together, these results suggest that infection with strain VGP would be an important causes of death in this giant panda cub.

ABSTRACT Tick infestations have been reported as one of the factors threatening the health of giant pandas, but studies of viral pathogens carried by ticks feeding on the blood of giant pandas are limited. To assess whether blood-sucking ticks of giant pandas can carry viral pathogens and if so, whether the viruses in ticks are associated with those previously detected in giant panda hosts, we determined the viromes of ticks detached from giant pandas in a field stocking area in Sichuan Province, southwest China. Using viral metagenomics we identified 32 viral species in ticks, half of which (including anellovirus [n = 9], circovirus [n = 3], and gemycircularvirus [n = 4]) showed homology to viruses carried by giant pandas and their associated host species (such as red pandas and mosquitoes) in the same living domain. Remarkably, several viruses in this study phylogenetically assigned as bunyavirus, hepe-like virus, and circovirus were detected with relatively high abundance, but whether these newly identified tick-associated viruses can replicate in ticks and then transmit to host animals during a blood meal will require further investigation. These findings further expand our understanding of the role of giant panda-infesting ticks in the local ecosystem, especially related to viral acquisition and transmission, and lay a foundation to assess the risk for giant panda exposure to tick-borne viruses. IMPORTANCE Ticks rank only second to mosquitoes as blood-feeding arthropods, capable of spreading pathogens (including viruses, bacteria, and parasites) to hosts during a blood meal. To better understand the relationship between viruses carried by ticks and viruses that have been reported in giant pandas, it is necessary to analyze the viromes of giant panda-parasitic blood-sucking ticks. This study collected 421 ticks on the body surface of giant pandas in Sichuan Province, China. We characterized the extensive genetic diversity of viruses harbored by these ticks and reported frequent communication of viruses between giant pandas and their ticks. While most of the virome discovered here are nonpathogenic viruses from giant pandas and potentially tick-specific viruses, we revealed some possible tick-borne viruses, represented by novel bunyaviruses. This research contributes to the literature because currently there are few studies on the virome of giant panda-infesting ticks.

The red panda is an endangered forest species distributed on the edge of the Qinghai Tibet Plateau. The species has been conserved in ex-situ in many countries and its survival is threatened by many diseases. Its immune system is vulnerable to age-associated alterations, which accumulate and result in a progressive deterioration that leads to an increased incidence of diseases. We identified 2,219 differentially expressed genes (DEGs) between geriatric (11–16 years) and adult individuals (4–8 years), and 1690 DEGs between adults and juveniles (1 year). The gene expression and functional annotation results showed that the innate immunity of red pandas increases significantly in geriatric individuals, whereas its change remains unclear when comparing adults and juveniles. We found that the adaptive immunity of red pandas first increased and then decreased with age. We identified CXCR3, BLNK, and CCR4 as the hub genes in the age-related protein–protein interaction network, which showed their central role in age-related immune changes. Many DNA repair genes were down-regulated in geriatric red pandas, suggesting that the DNA repair ability of the blood tissue in geriatric red pandas is significantly reduced. The significantly up-regulated TLR5 in geriatric individuals also suggests the possibility of enhancing the vaccination immune response by incorporating flagellin, which could be used to address decreased vaccine responses caused by age-related declines in immune system function. This work provides an insight into gene expression changes associated with aging and paves the way for effective disease prevention and treatment strategies for red pandas in the future.

Summary: Global biodiversity is facing serious threats. However, knowledge of the genomic consequences of recent rapid population declines of wild organisms is limited. Do populations experiencing recent rapid population decline have the same genomic status as wild populations that experience long-term declines? Yellow-breasted Bunting (Emberiza aureola) is a critically endangered species that has been experiencing a recent rapid population decline. To answer the question, we assembled and annotated the whole genome of Yellow-breasted Bunting. Furthermore, we found high genetic diversity, low linkage disequilibrium, and low proportion of long runs of homozygosity in Yellow-breasted Bunting, suggesting that the populations following recent rapid declines have different genomic statuses from the population that experienced long-term population decline.

Climate change-induced food shortages pose major threats to wildlife conservation, and the exclusive reliance of giant pandas on bamboo makes them particularly vulnerable. The aim of this study was to provide insight into the reasons for the foraging strategies of giant pandas to selectively forage for different bamboo parts (bamboo shoot, culm, and leaf) during different seasons. This study used a metabolomic approach to analyze the fecal metabolites of giant pandas and conducted a correlation analysis with their gut microbiota. The results indicate that the fecal metabolites of giant pandas differ significantly depending on the bamboo parts they forage on, with higher sugar content observed when they consume bamboo culm with high fiber content. By functional annotation, culm group metabolites were enriched in the galactose metabolic pathway, while shoot group metabolites were enriched in the phenylalanine, tyrosine and tryptophan biosynthesis pathways. Moreover, Streptococcus showed a significant positive correlation with glucose and acetic acid content. Therefore, the foraging strategy of giant pandas is based on the ability to utilize the nutrient content of different bamboo parts. Captive feeding and habitat construction should enrich bamboo species to allow them to express their natural foraging strategies and improve their welfare and reproductive status.

With the continuous development of business intelligence technology, the application research of decision support systems (DSSs) is deepening. In China, the work in this area started relatively late, and there are few DSS research cases to assist in marketing decision-making. Currently, marketing decision support systems have shortcomings in data integration, historical data, query functions, and data analysis. This article analyzes the characteristics of marketing decision-making, discusses the application of data warehouse, OLAP, and data mining technology in marketing decision support systems, and designs a marketing decision support system based on data mining technology. The system uses a BP neural network to conduct data mining marketing forecasting. A three-layer network model for marketing prediction is established, with sales time, product price, and customer purchasing power as network inputs and output as the sales volume of a certain type of product in different locations. The test results show that the average absolute percentage error of this method is 15.13%, and the prediction accuracy is high. Research shows that with the continuous development of data mining technology, the system cannot only help users conduct scientific and reasonable marketing decision-making analyses, making the marketing decision-making process more scientific and reasonable, but also can bring new ideas to enterprise decision-makers, promoting the continuous improvement and progress of the system.

Abstract Background The Chinese monal (Lophophorus lhuysii) is an endangered bird species, with a wild population restricted to the mountains in southwest China, and only one known captive population in the world. We investigated the fecal microbiota and metabolome of wild and captive Chinese monals to explore differences and similarities in nutritional status and digestive characteristics. An integrated approach combining 16S ribosomal RNA (16S rRNA) gene sequencing and ultra-high performance liquid chromatography (UHPLC) based metabolomics were used to examine the fecal microbiota composition and the metabolomic profile of Chinese monals. Results The results showed that the alpha diversity of gut microbes in the wild group were significantly higher than that in the captive group and the core bacterial taxa in the two groups showed remarkable differences at phylum, class, order, and family levels. Metabolomic profiling also revealed differences, mainly related to galactose, starch and sucrose metabolism, fatty acid, bile acid biosynthesis and bile secretion. Furthermore, strong correlations between metabolite types and bacterial genus were detected. Conclusions There were remarkable differences in the gut microbiota composition and metabolomic profile between wild and captive Chinese monals. This study has established a baseline for a normal gut microbiota and metabolomic profile for wild Chinese monals, thus allowing us to evaluate if differences seen in captive organisms have an impact on their overall health and reproduction.

Summary: Animals require specific blends of nutrients that vary across the life course and with circumstances, e.g., health and activity levels. Underpinning and complicating these requirements is that individual traits may be optimized on different dietary compositions leading to nutrition-mediated trade-offs among outcomes. Additionally, the food environment may constrain which nutrient mixtures are achievable. Natural selection has equipped animals for solving such multi-dimensional, dynamic challenges of nutrition, but little is understood about the details and their theoretical and practical implications. We present an integrative framework, nutritional geometry, which models complex nutritional interactions in the context of multiple nutrients and across levels of biological organization (e.g., cellular, individual, and population) and levels of analysis (e.g., mechanistic, developmental, ecological, and evolutionary). The framework is generalizable across different situations and taxa. We illustrate this using examples spanning insects to primates and settings (laboratory, and the wild), and demonstrate its relevance for human health.

Dental caries severely hinders efficient access to adequate energy in wildlife. Different food supplies will develop characteristic plaque, and the microorganisms of these plaque are closely related to dental health. Here, plaque samples from panda cubs with caries and caries-free were collected for 16S rRNA high-throughput sequencing. All sequences clustered into 337 operational taxonomic units (OTUs; 97% identity), representing 268 independent species belonging to 189 genera, 98 families, 51 orders, 24 classes, and 13 phyla. Two groups shared 218 OTUs, indicating the presence of a core plaque microbiome. α diversity analysis showed that the microbial diversity in plaques with caries exceeded that of caries-free. The dominant phyla of plaque microbiota included Proteobacteria, Bacteroidetes, Firmicutes, Fusobacteria, and Actinobacteria. The dominant genera included unclassified Neisseriaceae, Actinobacillus, Lautropia, Neisseria, Porhyromonas, unclassified Pasteurellaceae, Moraxella, Streptococcus, Bergeywlla and Capnocytophaga. β diversity analysis showed that the plaque microbial community structure was different between two groups. Using LEfSe analysis, 19 differentially abundant taxa were identified as potential biomarkers. Finally, function predictions analysis showed All the energy related metabolic pathways on KEGG level 2 were enriched in caries-active group. Consistent with the mainstream caries-causing narrative, our results illuminate the lack of information regarding the oral microflora composition and function within giant panda cubs.

In mammal herbivores, fiber digestion usually occurs predominantly in either the foregut or the hindgut. Reports of mechanisms showing synergistic function in both gut regions for the digestion of fiber and other nutrients in wild mammals are rare because it requires integrative study of anatomy, physiology, and gut microbiome. Colobine monkeys (Colobinae) are folivorous, with high-fiber foods fermented primarily in their foreguts. A few colobine species live in temperate regions, so obtaining energy from fiber during the winter is essential. However, the mechanisms enabling this remain largely unknown. We hypothesized that such species possess specialized mechanisms to enhance fiber digestion in the hindgut and studied microbial and morphological digestive adaptations of golden snub-nosed monkeys (GSMs), Rhinopithecus roxellana. which is a temperate forest colobine from central China that experiences high-thermal-energy demands while restricted to a fibrous, low-energy winter diet. We tested for synergistic foregut and hindgut fiber digestion using comparisons of morphology, microbiome composition and function, and digestive efficiency. We found that the GSM colon has a significantly greater volume than that of other foregut-fermenting colobines. The microbiomes of the foregut and hindgut differed significantly in composition and abundance. However, while digestive efficiency and the expression of microbial gene functions for fiber digestion were higher in the foregut than in the hindgut, both gut regions were dominated by microbial taxa producing enzymes to enable active digestion of complex carbohydrates. Our data suggest that both the GSM foregut and hindgut facilitate fiber digestion and that an enlarged colon is likely an adaptation to accommodate high throughput of fiber-rich food during winter.

Abstract Background Disease prevention and control is a significant part in the ex-situ conservation of the endangered red panda (Ailurus fulgens), being bacterial infection is one of the most important health threats to the captive population. To date, studies about the infection caused by Escherichia coli in the red panda are scarce. This study was conducted to determine the cause of death of a captive red panda through clinical symptoms, complete blood count, biochemical analysis, pathological diagnosis and bacterial whole genome sequencing. Case presentation The following report describes a case of a 1.5 year old captive red panda (Ailurus fulgens) that was found lethargic and anorectic. She was moved to the quarantine area for daily treatment with 50 mg of Cefpodoxime Proxetil. During the three-day treatment, she did not eat or defecate, and then died. Clinical hematology revealed the values of neutrophils, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and blood urea nitrogen (BUN) were significantly higher. Histological analysis demonstrated major pathological damage in the kidneys, liver and lungs, characterized by hyperemia, parenchymal cell degeneration and necrosis and inflammatory cell infiltration which were predominantly neutrophilic. A bacterial strain confirmed as Escherichia coli was isolated post mortem. Whole genome sequencing of the E. coli showed the complete genome size was 4.99 Mbp. PapA, PapC, OmpA, OmpU and other virulence factors which specific to Uropathogenic Escherichia coli (UPEC) were found in the isolate. Among the virulence factors, P pili, type I pili and related factors of the iron uptake system were associated with nephrotoxicity. Conclusion The red panda died of bacterial infection caused by an uropathogenic strain of Escherichia coli. The pathogenic mechanisms of the strain are closely related to the expression of specific virulence genes.

Abstract Background Parasitic infections are among the important causes of death of giant pandas (Ailuropoda melanoleuca) that hamper their survival in the wild. There are about 35 species of parasites which have been identified in giant pandas, but no information is currently available regarding the infection of Babesia in giant pandas. Babesia spp. are common intraerythrocytic parasite in wildlife, transmitted by ixodid ticks, which cause babesiosis. Clinical signs of babesiosis include fever, hemolysis, anemia, jaundice and death. Methods A species of Babesia was detected in the blood of a giant panda based on morphology and PCR amplification of the 18S rRNA gene. The phylogenetic relationship of Babesia sp. infecting giant panda was assessed by gene sequence alignment and phylogenetic analysis. Results Our analysis revealed that the Babesia isolate detected was most similar to an unidentified species of Babesia identified in black bears (Ursus thibetanus japonicus) from Japan (Babesia sp. Iwate, AB586027.1) with a 99.56% sequence similarity, followed by Babesia sp. EBB (AB566229.1, 99.50%) and Babesia sp. Akita (AB566229.1, 99.07%). Conclusions To our knowledge, this is the first report of Babesia detected in the giant panda. The results indicate that this Babesia sp. may be a novel species, currently named Babesia sp. strain EBP01.

The giant panda (Ailuropoda melanoleuca) is one of the world’s most endangered mammals and remains threatened as a result of intense environmental and anthropogenic pressure. The transformation and specialization of the giant panda’s diet into a herbivorous diet have resulted in unique adaptabilities in many aspects of their biology, physiology and behavior. However, little is known about their adaptability at the molecular level. Through comparative analysis of the giant panda’s genome with those of nine other mammalian species, we found some genetic characteristics of the giant panda that can be associated with adaptive changes for effective digestion of plant material. We also found that giant pandas have similar genetic characteristics to carnivores in terms of olfactory perception but have similar genetic characteristics to herbivores in terms of immunity and hydrolytic enzyme activity. Through the analysis of gene family expansion, 3752 gene families were found, which were enriched in functions such as digestion. A total of 93 genes under positive selection were screened out and gene enrichment identified these genes for the following processes: negative regulation of cellular metabolic process, negative regulation of nitrogen compound metabolic process, negative regulation of macromolecule metabolic process and negative regulation of metabolic process. Combined with the KEGG pathway, it was found that genes such as CREB3L1, CYP450 2S1, HSD11B2, LRPAP1 play a key role in digestion. These genes may have played a key role in the pandas’ adaptation to its bamboo diet.

Multi-drug-resistant Klebsiella pneumoniae (MDR K. pneumonia) is increasingly being reported with corresponding increase in morbidity and mortality all over the world. However, limited information is available concerning MDR K. pneumonia in giant pandas. The objective of this study was to grasp the drug resistance profile of MDR K. pneumonia isolated from giant pandas. A total of 182 K. pneumoniae isolates were collected from fresh feces of 94 captive giant pandas of different ages and sex and separated by season. We performed a standard disk diffusion antimicrobial susceptibility test with the isolates and further evaluated the antibiotic resistance genes (ARGs) of multi-drug-resistant strains by high-throughput quantitative PCR. In addition, we then analyzed mobile genetic elements (MGEs), integron gene cassettes, and the multi-locus sequence typing of multi-drug-resistant strains by PCR. Antimicrobial susceptibility testing results demonstrated that a total of 30 (16.5%) K. pneumoniae isolates showed multiple drug resistances. The thirty MDR K. pneumonia isolates were mainly resistant to amoxicillin (100.0%), doxycycline (86.7%), chloramphenicol (60.0%), compound trimethoprim (60.0%) and trimethoprim (56.7%). Fifty different types of antibiotic resistance genes were found, which included a total of 671 antibiotic resistance genes, in the 30 multi-drug-resistant isolates. The top ten resistance genes were: vanTC-02, aacC, blaCTX-M-04, blaSHV-01, blaSHV-02, ampC-04, blaOXY, tetD, blaTEM and tetA-02. Thirteen mobile genetic elements were detected, of which IS26 (96.67%) and intI1 (96.67%) had the highest frequency. The thirty MDR K. pneumonia isolates were negative for the traA, traF, tnsA, IS1133, ISpa7, ISkpn6, intI2 and intI3 genes. Moreover, a further investigation of integrons revealed that two types of specific gene cassettes (dfrA12 + orfF + aadA2 and dfrA12 + orfF) were identified in class 1 integrons. Multi-locus sequence typing results showed that 22 STs in the thirty MDR K. pneumonia isolates were identified, the main type was ST37 (5/30). Our results illustrate that effective surveillance and strict biosecurity strategies should be taken to prevent the spread of multi-drug-resistant bacteria, and monitor the emergence of mobile genetic elements and integrons.

A feline panleukopenia virus (FPV), Giant panda/CD/2018, was isolated from a captive giant panda with mild diarrhea in 2018 in Chengdu, China, and further identified via indirect immunofluorescence assay (IFA), transmission electron microscopy (TEM) observation, and genetic analysis. Phylogenetic analysis based on the complete VP2 nucleotide sequences showed that it shared high homology with Chinese FPV isolates and grouped within FPV cluster 1. One unique substitution Gly(G)299Glu(E) in the capsid protein VP2 was first identified with Giant panda/CD/2018. The presence of the G299E substitution is notable as it is located on the top region of the interconnecting surface loop 3, which may be involved in controlling the host range and antigenicity of FPV. These findings first demonstrate that FPV with natural point mutation G299E in the VP2 gene is prevalent in giant panda and suggest that etiological surveillance and vaccination among all giant pandas are urgently needed to protect this endangered species against FPV infection.

Deep neural networks (DNNs) with long-range dependence (LRD) have attracted more and more attention recently. However, LRD of DNNs is proposed from the view on gradient disappearance in training, which lacks theory analysis. In order to prove LRD of foggy images, the Hurst parameters of over 1,000 foggy images in SOTS are computed and discussed. Then, the Residual Dense Block Group (RDBG), which has additional long skips among two Residual Dense Blocks to fit LRD of foggy images, is proposed. The Residual Dense Block Group can significantly improve the details of dehazing image in dense fog and reduce the artifacts of dehazing image.

By analyzing the voiceprint characteristics of giant panda’s voice, this study proposes a giant panda individual recognition method based on the characteristics of the composite Mel composite frequency cepstral coefficient (CMFCC) and proves that the characteristic sequence of the CMFCC has long-range dependent characteristics. First, the MFCC (Mel composite frequency cepstral coefficient) with a low frequency resolution is obtained by the Mel filter bank; then, the inverse Mel frequency cepstral coefficient (IMFCC) features of giant panda calls are extracted. The CMFCC characteristic sequence of giant panda voice composed of the MFCC and IMFCC improves the resolution of high- and low-frequency resolution characteristics of giant panda voice. Finally, the first-order difference characteristic parameters of the MFCC are integrated to obtain the difference characteristics between frames. Through experiments, the improvement of the system recognition effect is verified, and the recognition accuracy meets the theoretical expectation.