Hematological and biochemical parameters of giant pandas (Ailuropoda melanoleuca) in captive and semi-natural environments.

Physiological indexes like blood parameters have been widely used to monitor the health of free-roaming animals. Attempts to reintroduce one of China's most endangered species, the giant panda (Ailuropoda melanoleuca), have been hampered by a lack of data on its ecology and physiology. We examined three giant pandas' hematological and blood chemistry parameters in a soft release program and 30 captive giant pandas as controls and determined the reference intervals (RIs) for those blood parameters in the captive animals. Elevation, captivity status and the interaction of those factors were statistically significant for hematologic measures. Release pandas had significantly higher hemoglobin and hematocrit values after they moved to high elevation locations. We also found significant difference in the enzyme parameters between high and low elevation pandas such as higher aspartate aminotransferase, alanine aminotransferase, creatinine kinase, amylase and lower lactate dehydrogenase and alkaline phosphatase. Release pandas also had higher nutrition parameter values such as higher albumin, globulin and creatinine. The RI for blood parameters in our study provides a baseline to monitor the health of captive animals and forms the basis for assessing the health of free-roaming giant pandas in future reintroduction efforts.

Measuring ecosystem services and ecological sensitivity for comprehensive conservation in Giant Panda National Park.

China announced the development of its first 5 national parks in 2021, the primary objective of which is to conserve the natural state and integrity of natural ecosystems. As such, ecosystem services and biodiversity levels are crucial assessment factors for the parks. For Giant Panda National Park (GPNP), we evaluated ecological sensitivity based on water and soil erosion and rocky desertification; ecosystem services based on headwater conservation, soil and water conservation, and biodiversity conservation; and presence of giant panda (Ailuropoda melanoleuca) and sympatric species (e.g., takin [Budorcas taxicolor], Asiatic black bear [Ursus thibetanus]) habitat suitability derived from niche modeling to identify the ecosystem status and assess ecological problems within the park. From our results, we proposed ecologically critical areas to target to meet the park's goals. The suitable habitat for pandas and sympatric species encompassed 62.98% of the park and occurred mainly in the Minshan Mountains. One quarter of the total area (25.67%) contained areas important for ecosystem services. Ecologically sensitive and extremely sensitive areas covered 88.78% of the park and were distributed mainly in Qionglaishan and Minshan Mountains. This coverage indicated that there was much habitat for pandas and sympatric species but that the ecosystems in GPNP are vulnerable. Therefore, ecologically critical areas encompassed all suitable habitats for all the species examined and areas important and extremely important to ecosystem service provision,ecologically sensitive and extremely sensitive areas, encompassed 15.17% of panda habitat, accounted for 16.37% of the GPNP area, and were distributed mainly in the Minshan Mountains. Our results indicated where conservation efforts should be focused in the park and that by identifying ecologically critical areas managers can provide targeted protection for wildlife habitat and ecosystems and effectively and efficiently protect the composite ecosystem. Additionally, our methods can be used to inform development of new national parks.

Medición de los servicios ambientales y la sensibilidad ecológica para una conservación integral en el Parque Nacional del Panda Gigante Resumen China anunció el crecimiento de sus primeros cinco parques nacionales en 2021, con el objetivo principal de conservar el estado natural y la integridad de los ecosistemas naturales. Para ello, los servicios ambientales y los niveles de biodiversidad son factores cruciales de evaluación para los parques. Para poder identificar el estado del ecosistema y evaluar los problemas ecológicos dentro del Parque Nacional del Panda Gigante (PNPG), analizamos la sensibilidad ecológica con base en la erosión del agua y del suelo y la desertificación rocosa; los servicios ambientales con base en el suministro de conservación del agua, del agua y del suelo y de la biodiversidad; y la idoneidad de hábitat del panda gigante (Ailuropoda melanoleuca) y de especies simpátricas (takín [Budorcas taxicolor], oso negro asiático [Ursus thibetanus]) derivada del modelo de nichos. A partir de nuestros resultados proponemos enfocarnos en áreas ecológicamente críticas para lograr los objetivos del parque. El hábitat idóneo para los pandas y las especies simpátricas englobó el 62.98% del parque y se ubicó principalmente en las montañas Minshan. Un cuarto del área total (25.67%) albergó áreas importantes para los servicios ambientales. Las áreas ecológicamente sensibles y extremadamente sensibles cubrieron el 88.78% del parque y se distribuyeron en las montañas Minshan y Qionglaishan. Esta cobertura indica que hay bastante hábitat para los pandas y las especies simpátricas pero que los ecosistemas en el PNPG son vulnerables. Por lo tanto, las áreas ecológicamente críticas englobaron todos los hábitats para todas las especies analizadas y todas las áreas importantes y extremadamente importantes para el suministro de servicios ambientales. Las áreas ecológicamente sensibles y extremadamente sensibles englobaron el 15.17% del hábitat del panda, representaron el 16.37% del área del PNPG y se localizaron principalmente en las montañas Minshan. Nuestros resultados indican en dónde se deben enfocar los esfuerzos de conservación dentro del parque y que, si identificamos las áreas ecológicamente críticas, los gestores pueden proporcionar una protección focalizada para el hábitat y los ecosistemas y así proteger efectiva y eficientemente el ecosistema compuesto. Además, nuestro método puede usarse para guiar el desarrollo de nuevos parques nacionales.

大熊猫国家公园能实现物种和生态完整性的多重保护 中国在2021年宣布设立首批5个国家公园, 保护自然生态系统的真实性和完整性是其优先目标, 而生态系统服务和生物多样性水平是关键的评估要素。大熊猫国家公园作为首批唯一以单一物种命名的国家公园, 通过评估其生态系统服务、生态敏感性, 同时结合大熊猫及同域分布物种(羚牛、亚洲黑熊)的栖息地适宜性, 以揭示其国家公园内生态系统的状态及其面临的问题, 明确其生态关键区以实现多重保护的目标。我们发现大熊猫国家公园包含了超过62.98%的大熊猫和同域物种的适宜栖息地, 主要分布在岷山山系;其次, 大熊猫国家公园包含了25.67%的生态系统服务重要区域和高达88.78%的生态敏感区域, 主要分布在岷山山系和邛崃山山系。这表明尽管大熊猫国家公园内包含了大熊猫及同域物种所需的大面积适宜栖息地, 但是其生态系统具有较强的脆弱性。若将同时包含大熊猫及同域物种的适宜栖息地、生态系统服务重要和极重要区、生态敏感和极敏感的区域定义为生态关键区, 其面积占比为16.37%, 覆盖了15.17%的大熊猫栖息地, 主要分布在岷山山系。因此, 基于生态关键区制定新的科学的、针对性的保护措施, 不仅可以更好的来保护野生动物栖息地和应对生态系统的威胁, 而且也有效且高效地保护多重生态系统。.

Zea mays genotype influences microbial and viral rhizobiome community structure.

Plant genotype is recognized to contribute to variations in microbial community structure in the rhizosphere, soil adherent to roots. However, the extent to which the viral community varies has remained poorly understood and has the potential to contribute to variation in soil microbial communities. Here we cultivated replicates of two Zea mays genotypes, parviglumis and B73, in a greenhouse and harvested the rhizobiome (rhizoplane and rhizosphere) to identify the abundance of cells and viruses as well as rhizobiome microbial and viral community using 16S rRNA gene amplicon sequencing and genome resolved metagenomics. Our results demonstrated that viruses exceeded microbial abundance in the rhizobiome of parviglumis and B73 with a significant variation in both the microbial and viral community between the two genotypes. Of the viral contigs identified only 4.5% (n = 7) of total viral contigs were shared between the two genotypes, demonstrating that plants even at the level of genotype can significantly alter the surrounding soil viral community. An auxiliary metabolic gene associated with glycoside hydrolase (GH5) degradation was identified in one viral metagenome-assembled genome (vOTU) identified in the B73 rhizobiome infecting Propionibacteriaceae (Actinobacteriota) further demonstrating the viral contribution in metabolic potential for carbohydrate degradation and carbon cycling in the rhizosphere. This variation demonstrates the potential of plant genotype to contribute to microbial and viral heterogeneity in soil systems and harbors genes capable of contributing to carbon cycling in the rhizosphere.

Centrifugation-Assisted Three-Dimensional Printing of Devices Embedded with Fully Enclosed Microchannels.

The challenges in reliably removing the sacrificial material from fully enclosed microfluidic channels hinder the use of three-dimensional (3D) printing to create microfluidic devices with intricate geometries. With advances in printer resolution, the etching of sacrificial materials from increasingly smaller channels is poised to be a bottleneck using the existing techniques. In this study, we introduce a microfabrication approach that utilizes centrifugation to effortlessly and efficiently remove the sacrificial materials from 3D-printed microfluidic devices with densely packed microfeatures. We characterize the process by measuring the etch rate under different centrifugal forces and developed a theoretical model to estimate process parameters for a given geometry. The effect of the device layout on the centrifugal etching process is also investigated. We demonstrate the applicability of our approach on devices fabricated using inkjet 3D printing and stereolithography. Finally, the advantages of the introduced approach over commonly used injection-based etching of sacrificial material are experimentally demonstrated in direct comparisons. A robust method to postprocess additively manufactured geometries composed of intricate microfluidic channels can help utilize both the large printing volume and high spatial resolution afforded by 3D printing in creating a variety of devices ranging from scaffolds to large-scale microfluidic assays.

Piglet cardiopulmonary bypass induces intestinal dysbiosis and barrier dysfunction associated with systemic inflammation.

The intestinal microbiome is essential to human health and homeostasis, and is implicated in the pathophysiology of disease, including congenital heart disease and cardiac surgery. Improving the microbiome and reducing inflammatory metabolites may reduce systemic inflammation following cardiac surgery with cardiopulmonary bypass (CPB) to expedite recovery post-operatively. Limited research exists in this area and identifying animal models that can replicate changes in the human intestinal microbiome after CPB is necessary. We used a piglet model of CPB with two groups, CPB (n=5) and a control group with mechanical ventilation (n=7), to evaluate changes to the microbiome, intestinal barrier dysfunction and intestinal metabolites with inflammation after CPB. We identified significant changes to the microbiome, barrier dysfunction, intestinal short-chain fatty acids and eicosanoids, and elevated cytokines in the CPB/deep hypothermic circulatory arrest group compared to the control group at just 4 h after intervention. This piglet model of CPB replicates known human changes to intestinal flora and metabolite profiles, and can be used to evaluate gut interventions aimed at reducing downstream inflammation after cardiac surgery with CPB.

Symbiotic microbiota and odor ensure mating in time for giant pandas.

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.

Field metabolic rates of giant pandas reveal energetic adaptations.

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.

Free-roaming dogs limit habitat use of giant pandas in nature reserves.

Giant pandas (Ailuropoda melanoleuca) were historically hunted using dogs and are currently threatened by free-roaming dogs and their associated diseases. To better understand the spatial magnitude of this threat, we used a GIS approach to investigate edge effects of dogs on giant panda habitat. We first examined two nature reserves with contrasting free-roaming dog populations: Liziping, with many dogs (~0.44/km2), and Daxiangling, with few dogs (~0.14/km2). Spatial analysis indicated that giant pandas at Liziping (but not Daxiangling) showed a shift in habitat use away from populated areas consistent with a risk response to the foray distance of free-roaming dogs (10.9 km path-distance). Most giant panda locations (86%) from the 2014 census in Liziping were clustered around remote "dog-free zones." Expanding this analysis across the entire giant panda range revealed that 40% of panda habitat is within the foray distance of dogs. Our assessment will inform dog control programs including monitoring, education, veterinary care, and other measures. We recommend that reserves designated for the release of translocated pandas receive priority consideration for dog control efforts. Only by understanding and managing complex interactions between humans, domestic animals, and wild animals can we sustain natural systems in a world increasingly dominated by humans.

Hybrid negative enrichment of circulating tumor cells from whole blood in a 3D-printed monolithic device.

Isolation and analysis of circulating tumor cells (CTCs) from blood samples present exciting opportunities for basic cancer research and personalized treatment of the disease. While microchip-based negative CTC enrichment offers both sensitive microfluidic cell screening and unbiased selection, conventional microchips are inherently limited by their capacity to deplete a large number of normal blood cells. In this paper, we use 3D printing to create a monolithic device that combines immunoaffinity-based microfluidic cell capture and a commercial membrane filter for negative enrichment of CTCs directly from whole blood. In our device, stacked layers of chemically-functionalized microfluidic channels capture millions of white blood cells (WBCs) in parallel without getting saturated and the leuko-depleted blood is post-filtered with a 3 µm-pore size membrane filter to eliminate anucleated blood cells. This hybrid negative enrichment approach facilitated direct extraction of viable CTCs off the chip on a membrane filter for downstream analysis. Immunofluorescence imaging of enriched cells showed ∼90% tumor cell recovery rate from simulated samples spiked with prostate, breast or ovarian cancer cells. We also demonstrated the feasibility of our approach for processing clinical samples by isolating prostate cancer CTCs directly from a 10 mL whole blood sample.

A novel polyomavirus from the nasal cavity of a giant panda (Ailuropoda melanoleuca).

BACKGROUND: Polyomaviruses infect a wide variety of mammalian and avian hosts with a broad spectrum of outcomes including asymptomatic infection, acute systemic disease, and tumor induction. METHODS: Viral metagenomics and general PCR methods were used to detected viral nucleic acid in the samples from a diseased and healthy giant pandas. RESULTS: A novel polyomavirus, the giant panda polyomavirus 1 (GPPyV1) from the nasal cavity of a dead giant panda (Ailuropoda melanoleuca) was characterized. The GPPyV1 genome is 5144 bp in size and reveals five putative open-reading frames coding for the classic small and large T antigens in the early region, and the VP1, VP2 and VP3 capsid proteins in the late region. Phylogenetic analyses of the large T antigen of the GPPyV1 indicated GPPyV1 belonged to a putative new species within genus Deltapolyomavirus, clustering with four human polyomavirus species. The GPPyV1 VP1 and VP2 clustered with genus Alphapolyomavirus. Our epidemiologic study indicated that this novel polyomavirus was also detected in nasal swabs and fecal samples collected from captive healthy giant pandas. CONCLUSION: A novel polyomavirus was detected in giant pandas and its complete genome was characterized, which may cause latency infection in giant pandas.

Role of New Nature Reserve in Assisting Endangered Species Conservation - Case Study of Giant Pandas in the Northern Qionglai Mountains, China.

The creation of nature reserves is the most direct way to save endangered species populations and their habitat. Development of the giant panda (Ailuropoda melanoleuca) nature reserve network in China was initiated in the 1960s, though the effort to create new reserves boomed considerably after the year 2000. Given this rapid development of protected areas in panda habitats, and the potential conflicting interests between conservation administrations and local economic development, it is essential to assess the role of new nature reserves in the overall giant panda conservation effort and reserve network. We utilized data from national giant panda surveys conducted in 2000 and 2012 to compare the size, spatial use, and distribution of panda populations, as well as the habitat suitability and connectivity in the Northern Qionglai Mountains between the two survey years. Our results show that although the total giant panda population in the study area did not change remarkably, local changes did occur. Most notably, the population in Wolong Nature Reserve declined by 27.3% (N = 39) and the population in Caopo Nature Reserve increased by 71.4% (N = 29) over the 12-year study period. We also found habitat suitability and availability decreased in both Wolong (12.4%) and Caopo (7.4%), but that the relative density of giant pandas declined (19.2%) and increased (84.6%) at each site, respectively. The distance between centers of high IUA were more distant in 2012 (14.1±1.9km) than that in 2000 (6.1±0.9km; t = -7.4, df = 5, p = 0.001), showing a scattered spatial pattern. Habitat availability decreased by 42% within the corridor between the two reserves, however panda occurrences in the corridor increased 24.6%. Compared to the total number of encounters, the proportion of the corridor increased 45.76%. Our results show the importance and success of the newly established Caopo to the conservation of giant pandas, and how crucial it is to identify and repair reserve corridors. Furthermore, we propose criteria for future nature reserve network management and investment, which is applicable for other endangered species conservation practices.

From frugivore to folivore: Altitudinal variations in the diet and feeding ecology of the Bioko Island drill (Mandrillus leucophaeus poensis).

Variation in the quality and availability of food resources can greatly influence the ecology, behavior, and conservation of wild primates. We studied the influence of altitudinal differences in resource availability on diet in wild drill monkeys (Mandrillus leucophaeus poensis) on Bioko Island, Equatorial Guinea. We compared fecal samples (n = 234) collected across three consecutive dry seasons for drills living in lowland (0-300 m asl) forest with nearby (18 km distance) drills living in montane forest (500-1000 m asl) in the Gran Caldera Southern Highlands Scientific Reserve. Lowland forest drills had a frugivorous diet very similar to that reported from studies on nearby mainland drills (M. l. leucophaeus) and mandrills (M. sphinx), with fruits comprising 90% of their dried fecal samples. However drills living in montane forest had a more folivorous diet, with herbaceous pith, leaves and fungi comprising 74% of their dried fecal samples and fruit becoming a minor component (24%). Furthermore, a dietary preference index indicated that the differences in the proportion of fruit and fibrous vegetation in the diets of lowland compared to montane drills was not simply a result of relative availability. Montane drills were actively consuming a higher mass of the available fruits and fibrous vegetation, a condition reflected in the greater mass of their fresh feces. Our results demonstrate the unexpected flexibility and complexity of dietary choices of this endangered species in two adjacent habitat types, a comparison of considerable importance for many other limited-range species faced with habitat loss and climate change.

Influence of saponins and tannins on intake and nutrient digestion of alkaloid-containing foods.

BACKGROUND: We hypothesized that eating a food containing saponins (SAP), or tannins (TAN) prior to foods containing the alkaloids gramine (GRA) or 5-methoxy-N,N-dimethyltryptamine (TRP) would provide benefits not possible when the alkaloid-containing foods were eaten alone. METHODS: In Trial 1, four groups of five lambs were first offered food with SAP for 30 min followed by food with either GRA or TRP for 3.5 h in a 2 × 2 factorial arrangement of a completely randomized design that included alkaloid (GRA or TRP) with or without SAP. In Trial 2 TAN replaced SAP. All foods were isocaloric (3.3 Mcal kg⁻¹) and isonitrogenous (14% crude protein). Foods, fecal and urine samples were collected and analyzed for dry matter intake and apparent digestibility of dry matter, energy (in megajoules, MJ), nitrogen (N), and neutral detergent fiber. RESULTS: Supplemental SAP did not affect digestibility of the parameters tested (P > 0.10). Supplemental TAN increased digestibility of N (g kg⁻¹, P = 0.04), N retained (g day⁻¹, P = 0.07), N digested (g day⁻¹, P = 0.06), and N retained/N consumed (g kg⁻¹, P = 0.07). However, digestibilities of dry matter (g kg⁻¹, P = 0.0026), energy (MJ 1000 MJ⁻¹, P = 0.003), neutral detergent fiber (g kg⁻¹, P = 0.008), and digested N retained (g kg⁻¹, P = 0.07) were lower for lambs fed TAN than for unsupplemented animals. CONCLUSIONS: Tannin supplementation can improve retention of nitrogen in animals fed alkaloid-containing grasses such as reed canarygrass and tall fescue. Combinations of forages with complementary primary and secondary compounds enable animals to maintain intake and improve nutrient utilization.

Supplementing endophyte-infected tall fescue or reed canarygrass with alfalfa or birdsfoot trefoil increases forage intake and digestibility by sheep.

BACKGROUND: We hypothesized that alfalfa (ALF) or birdsfoot trefoil (BFT) eaten prior to a meal of endophyte-infected tall fescue (TF) or reed canarygrass (RCG) would provide benefits not possible when TF or RCG are eaten alone. In trial 1, four groups of five lambs were first offered ALF for 30 min followed by either TF or RCG for 3.5 h in a 2 × 2 factorial arrangement of a completely randomized design that included grass (TF or RCG) with or without ALF. Trial 2 was similar to trial 1 except we used a new group of lambs fed BFT. Forage, fecal and urine samples were collected and analyzed for dry matter intake and apparent digestibility of dry matter, energy (in Kcal), nitrogen and neutral detergent fiber. RESULTS: A meal of either ALF or BFT prior to eating TF or RCG caused lambs to eat more total dry matter and nitrogen (both in g d⁻¹) and energy (in MJ d⁻¹) than lambs fed only TF or RCG (P < 0.05). Lambs fed ALF slightly reduced intake of TF and RCG relative to unsupplemented animals. Lambs fed BFT ate slightly less RCG, but much more TF than unsupplemented animals (P < 0.10). Lambs ate much less BFT than ALF, but they were stimulated to eat TF to a greater degree by BFT than by ALF. CONCLUSIONS: The enhanced intake of TF by lambs provided BFT, as well as the greater nutrient intake by lambs fed legumes and grasses was likely due in part to complementary profiles of alkaloids, saponins, and tannins.