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Griebenow, K., & Klibanov, A. M. (1995). Lyophilization-induced reversible changes in the secondary structure of proteins. Proc Natl Acad Sci USA, 92(24), 10969–10976.
Abstract: Changes in the secondary structure of some dozen different proteins upon lyophilization of their aqueous solutions have been investigated by means of Fourier-transform infrared spectroscopy in the amide III band region. Dehydration markedly (but reversibly) alters the secondary structure of all the proteins studied, as revealed by both the quantitative analysis of the second derivative spectra and the Gaussian curve fitting of the original infrared spectra. Lyophilization substantially increases the beta-sheet content and lowers the alpha-helix content of all proteins. In all but one case, proteins become more ordered upon lyophilization.
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Palme, R., Touma, C., Arias, N., Dominchin, M.N., & Lepschy, M. (2013). Steroid extraction: Get the best out of faecal samples. Wien Tierärztl Monat – Vet Med Austria, 100, 238–246.
Abstract: Faecal steroid hormone metabolites are becoming increasingly popular as parameters for reproductive functions and stress. The extraction of the steroids from the faecal matrix represents the initial step before quantification can be performed. The steroid metabolites present in the faecal matrix are of varying polarity and composition, so selection of a proper extraction procedure is essential. There have been some studies to address this complex but often neglected point. Radiolabelled steroids (e.g. cortisol or progesterone) have frequently been added to faecal samples to estimate the efficiency of the extraction procedures used. However, native, unmetabolized steroids are normally not present in the faeces and therefore the results are artifi- cial and do not accurately reflect the actual recoveries of the substances of interest. In this respect, recovery experiments based on faecal samples from radiometabolism studies are more informative. In these samples, the metabolite content accurately reflects the mixture of metabolites present in the given species. As a result, it is possible to evaluate different extraction methods for use with faecal samples. We present studies on sheep, horses, pigs, hares and dogs that utilized samples containing naturally metabolized, 14C-labelled steroids. We recommend extracting faecal steroids by simply suspending the faeces in a high percentage of a primary alcohol (for glucocorticoid metabolites 80% aqueous methanol proved best suited for virtually all mammalian species tested so far). Not only does the procedure significantly increase the total amount of recovered radioactivity, it also increases the percentage of unconjugated metabolites, which are more likely to be recognized by the antibodies used in various immunoassays. The advantages of this extraction procedure are clear: it is very easy to use (no evaporation step is needed), it yields high recoveries and variation based on the extraction procedure is reduced to a minimum.
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Gholib, G., Heistermann, M., Agil, M., Supriatna, I., Purwantara, B., Nugraha, T. P., et al. (2018). Comparison of fecal preservation and extraction methods for steroid hormone metabolite analysis in wild crested macaques. Primates, 59(3), 281–292.
Abstract: Since the non-invasive field endocrinology techniques were developed, several fecal preservation and extraction methods have been established for a variety of species. However, direct adaptation of methods from previous studies for use in crested macaques should be taken with caution. We conducted an experiment to assess the accuracy and stability of fecal estrogen metabolite (E1C) and glucocorticoid metabolite (GCM) concentrations in response to several preservation parameters: (1) time lag between sample collection and fecal preservation; (2) long-term storage of fecal samples in 80% methanol (MeOH) at ambient temperature; (3) different degrees of feces drying temperature using a conventional oven; and (4) different fecal preservation techniques (i.e., freeze-drying, oven-drying, and field-friendly extraction method) and extraction solvents (methanol, ethanol, and commercial alcohol). The study used fecal samples collected from crested macaques (Macaca nigra) living in the Tangkoko Reserve, North Sulawesi, Indonesia. Samples were assayed using validated E1C and GCM enzyme immunoassays. Concentrations of E1C and GCM in unprocessed feces stored at ambient temperature remained stable for up to 8 h of storage after which concentrations of both E1C and GCM changed significantly compared to controls extracted at time 0. Long-term storage in 80% MeOH at ambient temperature affected hormone concentrations significantly with concentrations of both E1C and GCM increasing after 6 and 4 months of storage, respectively. Drying fecal samples using a conventional oven at 50, 70, and 90 °C did not affect the E1C concentrations, but led to a significant decline for GCM concentrations in samples dried at 90 °C. Different fecal preservation techniques and extraction solvents provided similar results for both E1C and GCM concentrations. Our results confirm previous studies that prior to application of fecal hormone analysis in a new species, several preservation parameters should be evaluated for their effects on hormone metabolite stability. The results also provide several options for fecal preservation, extraction, and storage methods that can be selected depending on the condition of the field site and laboratory.
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Broekhuis, F., Madsen, E. K., & Klaassen, B. (2019). Predators and pastoralists: how anthropogenic pressures inside wildlife areas influence carnivore space use and movement behaviour. Anim Conserv, .
Abstract: Abstract Across the globe, wildlife populations and their behaviours are negatively impacted by people. Protected areas are believed to be an antidote to increasing human pressures but even they are not immune to the impact of anthropogenic activities. Areas that have been set aside for the protection of wildlife therefore warrant more attention when investigating the impact of anthropogenic pressures on wildlife. We use cheetahs Acinonyx jubatus as a case study to explore how a large carnivore responds to anthropogenic pressures inside wildlife areas. Using GPS-collar data we investigate cheetah space use, both when moving and stationary, and movement parameters (speed and turn angles) in relation to human disturbance, distance to human settlement, livestock abundance and livestock site use inside wildlife areas. Space use was negatively influenced by human disturbance, resulting in habitat loss and fragmentation and potentially reducing landscape permeability between neighbouring wildlife areas. Cheetahs were also less likely to stop in areas where livestock numbers were high, but more likely to stop in areas that were frequently used by livestock. The latter could reflect that cheetahs are attracted to livestock however, cheetahs in the study area rarely predated on livestock. It is therefore more likely that areas that are frequently used by livestock attract wild herbivores, which in turn could influence cheetah space use. We did not find any effects of people and livestock on cheetahs? speed and turn angles which might be related to the resolution of the data. We found that cheetahs are sensitive to human pressures and we believe that they could be an indicator species for other large carnivores facing similar challenges. We suggest that further research is needed to determine the levels of anthropogenic pressures needed to maintain ecological integrity, especially inside wildlife areas.
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Lee, P. (1991). Adaptation to environmental change:an evolutionary perspective. In H. O. Box (Ed.), Primate responses to environmental changes (pp. 39–56). London: Chapmann & Hall.
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Beck, B. B. (1980). Animal tool behaviour: The use and manufacture of tools by animals. New York: Garland.
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Fisher, J., & Hinde, R. A. (1994). The opening of milk bottles by birds. British Birds, (42), 347–357.
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Kuczaj, S. A., Makecha, R., Trone, M., Paulos, R. D., & Ramos, J. A. (2006). Role of Peers in Cultural Innovation and Cultural Transmission: Evidence from the Play of Dolphin Calves. Int. J. Comp. Psychol, 19(2), 223–240.
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Thorpe, W. H. (1963). Learning and Instinct in Animals. London: Methuen.
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Burke, C., Rashman, M., Wich, S., Symons, A., Theron, C., & Longmore, S. (2019). Optimizing observing strategies for monitoring animals using drone-mounted thermal infrared cameras. International Journal of Remote Sensing, 40(2), 439–467.
Abstract: ABSTRACTThe proliferation of relatively affordable off-the-shelf drones offers great opportunities for wildlife monitoring and conservation. Similarly the recent reduction in the cost of thermal infrared cameras also offers new promise in this field, as they have the advantage over conventional RGB cameras of being able to distinguish animals based on their body heat and being able to detect animals at night. However, the use of drone-mounted thermal infrared cameras comes with several technical challenges. In this article, we address some of these issues, namely thermal contrast problems due to heat from the ground, absorption and emission of thermal infrared radiation by the atmosphere, obscuration by vegetation, and optimizing the flying height of drones for a best balance between covering a large area and being able to accurately image and identify animals of interest. We demonstrate the application of these methods with a case study using field data and make the first ever detection of the critically endangered riverine rabbit (Bunolagus monticularis) in thermal infrared data. We provide a web-tool so that the community can easily apply these techniques to other studies (http://www.astro.ljmu.ac.uk/aricburk/uav_calc/).
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