@article{wood_minter_bibus_tollefson_ange-van heugten_2022, title={Assessment of the effects of storage temperature on fatty acid analysis using dried blood spot cards from managed southern white rhinoceroses (Ceratotherium simum simum): implications for field collection and nutritional care}, volume={10}, ISSN={2167-8359}, url={http://dx.doi.org/10.7717/peerj.12896}, DOI={10.7717/peerj.12896}, abstractNote={BackgroundSouthern white rhinoceroses (Ceratotherium simum simum) are an endangered species in decline due to poaching and negative habitat changes. Conservation of the species has become increasingly important and a focus on better human management has become prevalent. One area of management that impacts southern white rhinoceroses is nutritional health monitoring, which is often conducted through blood analysis. Blood analysis conducted during field research can be difficult due to temperature, distance, and limited technological resources, so new methods of fast, and relatively stable blood collection are being pursued. One method that has been used in humans for many years is beginning to make its way into wildlife studies: the use of dried blood spot (DBS) cards. These cards are used as a tool to store single drops of whole blood on specialized filter paper and, once dried, can be used for nutritional biomarker analysis. An area of interest for southern white rhinoceroses and nutrition is monitoring fatty acid percentages for cardiovascular, immune, and reproductive health. The time and temperature limitations for storing blood fractions or liquid whole blood when analyzing fatty acids have been investigated, but few studies have performed storage studies on DBS cards colder than −20 °C or in non-human species.MethodsIn order to better understand the limitations of DBS cards and the impact of temperature on fatty acid DBS samples in long-term storage, triplicate samples from seven adult southern white rhinoceroses at the North Carolina Zoo were collected and subjected to three storage treatments (immediate, room temperature (23 °C), or frozen (−80 °C) for 1 year).ResultsStearidonic (18:4w3) (Δ 0.3%), arachdic (20:0) (Δ 0.1%), eicosatetraenoic (20:4w3) (Δ 0.2%), and erucic acid (22:1w9) (Δ 0.1%) were in higher concentration in frozen than initial. Fatty acids in higher concentrations in the initial samples than frozen were myristic (14:0) (Δ 0.2%), mead (20:3w9) (Δ 0.1%), docosatetraenoic (22:4w6) (Δ 0.2%), nervonic (24:1) (Δ 0.1%), and total highly unsaturated fatty acids (HUFAs) (Δ 0.7%). Stearic (18:0) (Δ 2.2%), stearidonic (18:4w3) (Δ 0.3%), arachdic (20:0) (Δ 0.2%), paullinic (20:1w7) (Δ 0.4%), eicosatetraenoic (20:4w3) (Δ 0.1%), eicosapentaenoic (20:5w3) (Δ 0.1%), docosatetraenoic (22:4w6) (Δ 0.2%), nervonic acid (24:1) (Δ 0.2%), monoenes (Δ 1.9%), and total saturates (Δ 3.6%) had higher concentrations in room temperature than initial. Linoleic (18:2w6) (Δ 4.9%), mead acid (20:3w9) (Δ 0.1%), total polyunsaturated fatty acids (5.3%), and total omega-6 fatty acids (Δ 4.8%) had higher concentrations in initial compared to room temperature. Arachidonic (20:4w6) (Δ 0.4%) and omega-3 docosapentaenoic acid (22:5w3) (Δ 0.1%), had higher concentrations in frozen than in room temperature.DiscussionThe frozen samples had the fewest statistical differences compared to room temperature samples and essential omega-3 and -6 fatty acids were stable with freezing up to 1 year. While more research is still warranted, current results suggest that DBS samples are best utilized when immediate analysis or −80 °C storage is available.}, journal={PeerJ}, publisher={PeerJ}, author={Wood, Jordan and Minter, Larry J. and Bibus, Doug and Tollefson, Troy N. and Ange-van Heugten, Kimberly}, year={2022}, month={Feb}, pages={e12896} }
 @article{wood_morgan_ange-van heugten_serrano_minter_fellner_stoskopf_2022, title={Observable Metabolites and Metabolomic Sampling Protocols for Managed African Savanna Elephant (Loxodonta africana) Whole Blood Using H-NMR Spectroscopy}, volume={12}, ISSN={["2218-1989"]}, url={https://doi.org/10.3390/metabo12050400}, DOI={10.3390/metabo12050400}, abstractNote={We used nuclear magnetic spectroscopy (NMR) to evaluate the metabolomics of heparinized whole blood drawn from six African savanna elephants (Loxodonta africana) maintained on a well characterized diet. Whole blood samples obtained under behavioral restraint, then quickly frozen in liquid nitrogen, were stored at −80 °C until analysis. Frozen samples were thawed under controlled conditions and extracted with methanol and chloroform to separate the polar and non-polar metabolites. We identified 18 polar metabolites and 14 non-polar lipids using one-dimensional (1D) and two-dimensional (2D) NMR spectra. Despite unexpected rouleaux formation in the thawed frozen samples, spectra were consistent among animals and did not vary dramatically with age or the sex of the animal.}, number={5}, journal={Metabolites}, author={Wood, J. and Morgan, D.R. and Ange-van Heugten, K. and Serrano, M. and Minter, L.J. and Fellner, V. and Stoskopf, M.K.}, year={2022}, pages={400} }
 @article{trivedi_burnham_capobianco_boshoff_zheng_pettiglio_ange-van heugten_bissell_minter_2021, title={Analysis of Blood Biochemistry of Free Ranging and Human-Managed Southern White Rhinoceros (Ceratotherium simum simum) Using the i-STAT Alinity v®}, volume={2021}, ISSN={2042-0048 2090-8113}, url={http://dx.doi.org/10.1155/2021/2665956}, DOI={10.1155/2021/2665956}, abstractNote={Handheld point-of-care blood analyzers deliver rapid results for biochemical and hematologic parameters, making them very useful in veterinary clinics and in fieldwork applications. This study compared the biochemical and hematologic parameters generated by the novel point-of-care analyzer i-STAT® Alinity V CHEM8+ cartridge between human-managed and free ranging populations of southern white rhinoceros (Ceratotherium simum simum). In addition, a novel reference interval for ionized calcium (iCa), a parameter of diagnostic and prognostic importance, was established for southern white rhinoceros. Blood samples were obtained from 10 managed (6 at NC Zoo and 4 at Busch Gardens Tampa Bay in 2019) and 30 free ranging white rhinoceros (collected in South Africa between 2018 and 2019) and analyzed using the i-STAT. Multiple parameters were higher (
                     
                        P
                        <
                        0.05
                     
                  ) in free ranging versus managed animals including potassium, blood urea nitrogen, creatinine, glucose, hematocrit, and hemoglobin. Conversely, iCa concentrations were higher (
                     
                        P
                        <
                        0.05
                     
                  ) in the managed populations of white rhinoceros. The RI determined for iCa was 1.36–1.56 mmol/L, with a mean of 1.46 mmol/L, and was determined using the guidelines from the American Society for Veterinary Clinical Pathology. There was no difference in anion gap, chlorine, total carbon dioxide, or sodium between the populations. Seasonality and locality of sampling as well as diet may be contributing factors to the higher iCa concentrations in managed rhinoceros. The six elevated parameters in free ranging rhinoceros are likely attributable to dehydration compounded by capture stress. This data provides insight into the state of several biochemical and hematologic parameters in southern white rhinoceros and will allow veterinarians to better assess the health of both managed and free ranging populations.}, journal={Veterinary Medicine International}, publisher={Hindawi Limited}, author={Trivedi, Shweta and Burnham, Christina M. and Capobianco, Christian M. and Boshoff, Christiaan and Zheng, Yaxin and Pettiglio, Jordan Wood and Ange-van Heugten, Kimberly and Bissell, Heidi D. and Minter, Larry J.}, editor={Hussni, Carlos AlbertoEditor}, year={2021}, month={Jul}, pages={1–6} }
 @article{wood_minter_tollefson_bissell_bibus_ange-van heugten_2021, title={Implications of Nutritional Management on Fatty Acid Profiles of Southern White Rhinoceroses (Ceratotherium simum simum) Housed at Two Zoological Institutions}, volume={11}, ISSN={2076-2615}, url={http://dx.doi.org/10.3390/ani11113063}, DOI={10.3390/ani11113063}, abstractNote={Southern white rhinoceroses (Ceratotherium simum simum) are African megaherbivores that are considered near threatened by the International Union for the Conservation of Nature. The fatty acid circulating values of these animals have not been thoroughly investigated. Fatty acids are critical for immune, heart, skin, and reproductive health, and may have a significant impact on the management and conservation of this species. Published data on fatty acids in this species is limited to incomplete profiles with very few animals in managed environments. The objectives of this research were to provide novel fatty acid percentage profiles for managed healthy southern white rhinoceroses, as well as to provide comparisons between two zoological institutions with differences in diet and climate during two distinct pasture growth periods. Whole blood samples were collected as dried blood spots from six rhinoceroses at the North Carolina Zoo (NC Zoo) and five rhinoceroses at Busch Gardens Tampa (BGT) in the low growth period (February to April) of 2019 and during the high growth period (July to September) of 2020. Fatty acid results indicated numerous differences when comparing the institutions within the same growth period and when comparing the same institution between its two growth periods. Most noteworthy were the higher levels of α-linolenic acid (18:3w3) and total omega-3 fatty acids and the lower linoleic acid (18:2w6), total omega-6 fatty acids, and omega-6 to omega-3 ratio found in the BGT population in both growth periods. This study provides novel percentages of fatty acids in managed southern white rhinoceroses and data on how fatty acid profiles may be altered between two housing locations via dietary differences in hay type and quantity, pasture availability via season, and pellet inclusion levels.}, number={11}, journal={Animals}, publisher={MDPI AG}, author={Wood, Jordan and Minter, Larry and Tollefson, Troy Neil and Bissell, Heidi and Bibus, Doug and Ange-van Heugten, Kimberly}, year={2021}, month={Oct}, pages={3063} }
 @article{wood_koutsos_kendall_minter_tollefson_ange‐van heugten_2020, title={Analyses of African elephant (
            Loxodonta africana
            ) diet with various browse and pellet inclusion levels}, volume={39}, ISSN={0733-3188 1098-2361}, url={http://dx.doi.org/10.1002/zoo.21522}, DOI={10.1002/zoo.21522}, abstractNote={AbstractTo more closely simulate the diet of free‐ranging elephants, the diet of six (2.4) African elephants (Loxodonta africana) was altered to include more browse and less pelleted complete feed (5% total diet). Dietary proximate compounds, minerals, vitamins A (and carotenoids), D and E, and fatty acids were analyzed on pelleted diet items and forages including hay, grass, and browse. A total of 42 browse species were offered over 1 year with an average total diet inclusion of 5.2% (dry matter basis) per day. Dietary Na and Se were low while Fe and Mn were high compared to published intake levels for elephants. Analyzed nutrients within browse varied widely among seasons and species. Ingredient analyses were used to create predicted elephant nutrient intake for (a) the current diet, (b) a diet excluding pellets, and (c) a diet excluding pellets and providing browse at doubled levels. Formulated diets excluding pellets had lower mineral levels than the current diet and doubled browse did not alter mineral inclusions of concern. This study provides seasonal data on the nutrient levels of Southeastern browse species important for various pachyderm and herbivorous species. Predicted nutrient intake with new diet scenarios does not support the exclusion of pellets in the diets of African elephants without greater browse quantity availability, strict diet management, or additional supplements.}, number={1}, journal={Zoo Biology}, publisher={Wiley}, author={Wood, Jordan and Koutsos, Elizabeth and Kendall, Corinne J. and Minter, Larry J. and Tollefson, Troy N. and Ange‐van Heugten, Kimberly}, year={2020}, month={Jan}, pages={37–50} }