Hair cortisol analyses in different mammal species: choosing the wrong assay may lead to erroneous results

Abstract Wild animals are faced with a broad range of environmental stressors and research is needed to better understand their effect on populations. Hormone analysis based on enzyme immunoassays (EIAs) can provide valuable information on adrenocortical activity (stress), and assessment of cortisol in hair may allow the quantification of cortisol production. To validate hair hormone analysis, we compared two EIAs based on antibodies against cortisol-3-CMO-BSA and cortisol-21-HS-BSA for hair glucocorticoid (hGC) measurements in Egyptian mongoose, Iberian lynx, Alpine marmot, Asiatic black bear, spotted hyena and cheetah, with results obtained by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) measurements. Both EIAs were also characterized by HPLC immunograms. Our results revealed that the cortisol-21-HS EIA measured 2.3- to 12-fold higher hGC concentrations than the cortisol-3-CMO assay. In dependence of the species, high-performance liquid chromatography (HPLC) immunograms showed that up to 70% of immunoreactivities determined by the cortisol-21-HS constituted of unknown unpolar compounds leading to an overestimation of hGC. The cortisol-3-CMO EIA expressed a better specificity, with 32.1–67.4% of immunoreactivity represented by cortisol and cortisone. The LC-MS/MS analyses (gold standard) revealed that the cortisol-3-CMO EIA also resulted in an (up to 3-fold) overestimation of hGC, but EIA results were correlated with LC-MS/MS in the mongoose, the lynx, the spotted hyena and the marmot. No correlation was obtained for Asiatic black bears. As a result of our study, we strongly recommend to test any cortisol EIA for its specificity towards extracted hair components. In all analyzed species, except the Asiatic black bear, cortisone and cortisol were simultaneously present in hair extracts; consequently, an appropriate EIA should cross-react to these two glucocorticoid hormones and express negligible affinity towards substances with less polarity than corticosterone. Choosing the wrong EIA for hGC analyses may lead to overestimations of hGC or—in the worst case—to results that do not mirror real adrenocortical activity.

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Physicochemical stability of the bevacizumab biosimilar, ABP 215, after preparation and storage in intravenous bags

Generics and Biosimilars Initiative Journal ◽

10.5639/gabij.2020.0904.026 ◽

2020 ◽

Vol 9 (4) ◽

pp. 155-162

Author(s):

Jolita Seckute ◽

Ingrid Castellanos ◽

Steven Bane

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

High Performance ◽

Protein Concentration ◽

Sodium Dodecyl ◽

Significant Loss ◽

Size Exclusion ◽

Worst Case ◽

Physicochemical Stability ◽

And Storage

Study Objectives: To evaluate extended in-use stability of bevacizumab biosimilar, ABP 215, after dilution into intravenous bags, extended storage, and simulated infusion to enable advanced preparation and storage. Methods: Two lots of ABP 215 were diluted to high- (16.5 mg/mL) and low- (1.4 mg/mL) dose concentrations in two types of intravenous bag under ambient light conditions. Dosed intravenous bags were stored at 2°C–8°C for 35 days, followed by 30°C for 2 days, and each bag was infused on Day 37. Analysis of purity and physicochemical stability was performed using size-exclusion high-performance liquid chromatography (SE-HPLC), cation-exchange high-performance liquid chromatography (CEX-HPLC), reduced capillary electrophoresis-sodium dodecyl sulphate (rCE-SDS), subvisible particle detection assays, visual inspection, and by measuring protein concentration and potency. Results: No meaningful changes were seen in ABP 215 purity when analysed by SE-HPLC, CEX-HPLC and rCE-SDS following dilution, storage and infusion of two lots, bags, and doses. Protein concentration remained consistent throughout the study for all samples and no significant loss in potency was detected. No potentially proteinaceous particles or increases in subvisible particles were observed. Discussion: This study investigated the in-use stability of ABP 215 following dilution, extended storage, and infusion, that represent worst-case handling conditions. ABP 215 exhibited consistent product quality and activity, with no significant degradation observed under the conditions tested. Conclusion: ABP 215 retains physicochemical stability after dilution over the recommended dosing concentrations, extended storage, and simulated infusion. This supports the advance preparation and storage of ABP 215 in intravenous bags for infusion.

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Mammals of Tengchong Section of Gaoligongshan National Nature Reserve in Yunnan Province, China

Journal of Threatened Taxa ◽

10.11609/jott.4439.11.11.14402-14414 ◽

2019 ◽

Vol 11 (11) ◽

pp. 14402-14414 ◽

Cited By ~ 1

Author(s):

Fei Li ◽

Xiang-Yuan Huang ◽

Xing-Chao Zhang ◽

Xing-Xi Zhao ◽

Jian-Huan Yang ◽

...

Keyword(s):

Yunnan Province ◽

Nature Reserve ◽

Black Bear ◽

Iucn Red List ◽

Livestock Grazing ◽

Mammal Species ◽

Field Surveys ◽

Asiatic Black Bear ◽

National Nature Reserve ◽

Low Altitude

We conducted field surveys on the mammalian diversity in the Tengchong Section of Gaoligongshan National Nature Reserve in Yunnan Province, China, using camera trap and transect method between April 2014 and May 2018. A total of 46 identifiable mammal species were recorded, including one new record for China and nine new species for the Tengchong County. Of the 46 species, nine are globally threatened (three Endangered and six Vulnerable) and six are Near Threatened on the IUCN Red List of Threatened Species. Species richness of most mammal groups in Tengchong remains high compared to similar sites in neighbouring countries; however, encounter rates for species vulnerable to high hunting pressures were quite low and large carnivores, except the Asiatic Black Bear, were either extirpated or at critically low numbers. Future surveys should cover a wider elevation range and a variety of microhabitats to increase the probability of detecting the species not yet recorded. To enhance the conservation value of the reserve for the diverse mammalian community, poaching and livestock grazing should be further controlled; in addition, protection and restoration of low-altitude forests should be encouraged. Monitoring and research on selected flagship species should also be conducted.

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Mammals of the Bhagirathi basin, Western Himalaya: understanding distribution along spatial gradients of habitats and disturbances

Oryx ◽

10.1017/s0030605319001352 ◽

2020 ◽

pp. 1-11

Author(s):

Ranjana Pal ◽

Shagun Thakur ◽

Shashank Arya ◽

Tapajit Bhattacharya ◽

Sambandam Sathyakumar

Keyword(s):

Anthropogenic Impacts ◽

Brown Bear ◽

Black Bear ◽

Lynx Lynx ◽

Panthera Tigris ◽

Anthropogenic Pressures ◽

Mammal Species ◽

Eurasian Lynx ◽

Asiatic Black Bear ◽

Cuon Alpinus

Abstract Understanding the distribution of wildlife species and their response to diverse anthropogenic pressures is important for conservation planning and management of wildlife space in human-dominated landscapes. Assessments of anthropogenic impacts on mammals of the Indian Himalayan Region have mostly been limited to locations inside protected areas. We studied the occurrence of mammals in an unexplored landscape, the 7,586 km2 Bhagirathi basin, at an altitude of 500–5,200 m. The basin encompasses wilderness areas of various habitat types and protection status that are exposed to a range of anthropogenic pressures. Camera trapping at 209 locations during October 2015–September 2017 confirmed the occurrence of 39 species of mammals, nine of which are categorized as threatened (four Vulnerable, five Endangered) and four as Near Threatened on the IUCN Red List. We recorded five mammal species that were hitherto undocumented in Uttarakhand State: the argali Ovis ammon, Tibetan sand fox Vulpes ferrilata, woolly hare Lepus oiostolus, Eurasian lynx Lynx lynx and woolly flying squirrel Eupetaurus cinereus. In addition, we recorded two Endangered species, the dhole Cuon alpinus and tiger Panthera tigris. Threatened species such as the sambar Rusa unicolor, common leopard Panthera pardus and Asiatic black bear Ursus thibetanus occur in a wide variety of habitats despite anthropogenic disturbance. We recorded the snow leopard Panthera uncia in areas with high livestock density but temporally segregated from human activities. The musk deer Moschus spp. and Himalayan brown bear Ursus arctos isabellinus were recorded in subalpine habitats and appeared to be less affected by human and livestock presence. Our findings highlight the potential of the Bhagirathi basin as a stronghold for conservation of several threatened and rare mammal species.

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