Analysis of Fatty Acid Profiles for Eastern Box (Terrapene Carolina Carolina) and Common Snapping (Chelydra Serpentine) Turtles in Wild and Managed Care Environments

Veterinary Medicine International

Stoskopf

et al. 2021

Self Cite

Fatty acids, especially omega-3 and omega-6 fatty acids, are important for reproductive and cardiovascular health in animals. While monitoring fatty acids is traditionally conducted using frozen blood fractions such as serum and plasma, advancements in analytical technology have developed a method of collecting microsamples of dried whole blood on Ahlstrom 226 grade filter paper that can provide information on long-term fatty acid status of animals. Blood samples were collected from five male pigs in both the traditional frozen method and on dried blood spot cards (DBS). The DBS samples were collected with untreated syringes and tubes, and approximately 320 μL of blood was placed on each card with approximately 80 μL per spot (4 spots). Statistical analysis was performed to compare the two sample groups to each other using the Mann–Whitney U-test and determine if DBS samples were similar to traditional whole blood samples. Of the 30 fatty acids and fatty acid groups with measurable concentrations, only four individual fatty acids, behenic acid, omega-3 docosapentaenoic acid, nervonic acid, and adrenic acid, had statistical differences. Most of these differences were minor and could be due to analytical errors or contamination. Comparisons between sample types found similar concentrations of key omega fatty acids and PUFAs and support the use of DBS collection as a less invasive method of blood collection and fatty acid analysis.

Section: Discussionmentioning

confidence: 99%

Investigation of Dried Blood Spot Cards for Fatty Acid Analysis Using Porcine Blood

Veterinary Medicine International

Stoskopf

et al. 2021

Self Cite

Section: Introductionmentioning

confidence: 99%

“…While used since the early 1920s in human health monitoring ( Freeman et al, 2018 ), dried blood spot (DBS) filter cards have begun to be used in field research for animals, including fatty acid analysis ( Brindle, O’Connor & Garrett, 2014 ; Dass et al, 2020 ; Koutsos et al, 2021 ; Wood et al, 2021a ; Wood et al, 2021b ; Wood et al, 2021c ). This work has been conducted in multiple reptile species ( Dass et al, 2020 ; Koutsos et al, 2021 ) but validation work to determine if DBS can be directly compared to liquid whole blood in the domestic pig has also been conducted ( Wood et al, 2021b ). Dried blood spot samples are an important alternative to liquid or frozen whole blood or blood fractions for field work collection because they do not require large amounts of blood to be drawn, only require a few drops of whole blood on specialized filter paper for analysis and potentially have minimal storage needs (no immediate processing or freezing) ( Freeman et al, 2018 ).…”

Section: Introductionmentioning

confidence: 99%

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

Bibus

et al. 2022

PeerJ

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Background Southern 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. Methods In 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). Results Stearidonic (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. Discussion The 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.

“…While understudied in other species, correlations between dietary omega-3 and omega-6 imbalances and atherosclerosis have been noted in several species, including African elephants ( McCullaugh, 1972 ). Additionally, studies have found differences in circulating fatty acids when comparing free ranging versus managed animals for several species ( Clauss et al, 2008 ; Clauss, Grum & Hatt, 2007 ; Dass et al, 2020 ; Dass et al, 2021 ; Schmidt et al, 2009 ).…”

Section: Introductionmentioning

confidence: 99%

“…These only require small volumes of whole blood to run full free fatty acid profiles and are more easily collected and stored in field research settings ( Freeman et al, 2018 ). Because of this positive impact on field research, DBS cards have become more prevalent in wildlife research, but direct comparisons to serum, plasma, or liquid whole blood for a majority of species is lacking ( Koutsos et al, 2021 ; Dass et al, 2020 ; Dass et al, 2021 ). This has led to the research question of how comparable DBS samples compare to more traditional samples such as liquid whole blood, serum, or plasma regarding fatty acid profiling.…”

Section: Introductionmentioning

confidence: 99%

Comparison of African savanna elephant (Loxodonta africana) fatty acid profiles in whole blood, whole blood dried on blood spot cards, serum, and plasma

Bibus

et al. 2021

PeerJ

Self Cite

Background African elephants in managed care have presented differences in the balance between omega-3 and omega-6 fatty acids, a situation primarily thought to be due to dietary differences between the managed animals and their free-ranging counterparts. Because of this, circulating fatty acid status is included in routine monitoring of elephant health. A method of blood collection that requires only a few drops of whole blood, dried on filter paper (DBS) and can be used for analyzing full fatty acid profiles offers advantages in clinical application. Methods This study compared the use of whole blood, and whole blood DBS, serum or plasma for use in evaluating circulating fatty acid composition in African savannah elephants. Samples from six African elephants (two males and four females) were collected during the same week at the NC Zoo, Asheboro, NC. Results Results found only 2 of 36 individual fatty acids and none of the 10 fatty acid groupings were different when comparing the four blood fraction sample types to each other with Mann-Whitney U-Test pairwise comparisons. Myristic acid (14:0) was lower in the DBS samples than in whole blood, serum, and plasma and pentadecaenoic acid (15:1) was slightly more concentrated in DBS and whole blood. Discussion Results indicate that fatty acid profile of serum, plasma, whole blood, and DBS are comparable in African elephants. The DBS method offers advantages in acquisition and handling and may be preferable to other methods in both routine health assessment of captive animals and field research on free ranging animals.