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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

This article describes the deuterium oxide dilution technique in two mammals, an insectivore and carnivore, to determine total body water, lean body mass, body fat mass, and water consumption.

Abstract

Body condition scoring systems and body condition indices are common techniques used for assessing the health status or fitness of a species. Body condition scoring systems are evaluator dependent and have the potential to be highly subjective. Body condition indices can be confounded by foraging, the effects of body weight, as well as statistical and inferential problems. An alternative to body condition scoring systems and body condition indices is using a stable isotope such as deuterium oxide to determine body composition. The deuterium oxide dilution method is a repeatable, quantitative technique used to estimate body composition in humans, wildlife, and domestic species. Additionally, the deuterium oxide dilution technique can be used to determine the water consumption of an individual animal. Here, we describe the adaption of the deuterium oxide dilution technique for assessing body composition in big brown bats (Eptesicus fuscus) and for assessing water consumption in cats (Felis catis).

Introduction

Body condition scoring systems and body condition indices are common techniques used for assessing the health status or fitness of a species1,2. Many domestic and zoological species have unique body condition scoring (BCS) systems that are used to assess an animal's muscle and superficial fatty tissue3. However, BCS assessment relies upon the evaluator—meaning that BCS is an objective or semiquantitative measurement when assessed by a trained evaluator. In wildlife species, body condition indices are commonly used rather than BCS and are based upon a ratio of body mass to body....

Protocol

All experiments described here were approved by the University of Missouri Animal Care and Use Committee and conducted under the Missouri Department of Conservation (MDC) Wildlife Scientific Collection permit (Permit #16409 and #17649).

1. Preparation of sterile, isotonic, salinated D2O stock solution

  1. Make a 50 mL stock solution of 9.0 g/L salinated D2O.
    1. Weigh 450 mg of pharmaceutical grade NaCl and transfer all NaCl into a 100 mL, sterilized beake.......

Representative Results

The deuterium oxide dilution technique can be used to assess the body composition of a variety of species. To demonstrate the adaptability, we are reporting the first use of the deuterium oxide dilution technique in a North American insectivorous bat species, Eptesicus fuscus, the big brown bat for representative results. A timing plateau was completed by taking pre- and post-D2O injection blood samples as should be done with any species where the equilibration period .......

Discussion

The use of deuterium oxide to determine TBW has been used since the 1940s17 and is used in humans and a variety of domestic and wildlife species4,6,7. Other non-destructive techniques have been developed including bioelectrical impedance analysis (BIA), DXA, and quantitative magnetic resonance (QMR). Each method has advantages and disadvantages that should be considered before selecting a particular metho.......

Acknowledgements

This research was supported by MDC Cooperative Agreement (#416), US Forest Service Cooperative Agreement (16-JV-11242311-118), American Academy of Veterinary Nutrition and Waltham/Royal Canin, USA Grant (grant number: 00049049), NIH training grant (grant number: T32OS011126), and the University of Missouri Veterinary Research Scholars Program. The authors thank Shannon Ehlers for pre-reviewing this manuscript. We thank Dr. Robert Backus for providing the D2O standards and allowing use of his laboratory.

....

Materials

NameCompanyCatalog NumberComments
0.2 micron non-pyrogenic disk filterArgos TechnologiesFN32Snylon, 30mm diameter, 0.22um, sterile
1.5 mL conical microcentrifuge tubesUSA Scientific1415-97011.5 ml self-standing microcentrifuge tube, natural with blue cap
10 mL sterile glass vial for injectionMountainside Medical EquipmentMS-SEV10clear, sterile glass injection unit
10 mL syringeBecton Dickinson305219sterile 10 mL syringe individually wrapped
100 mL sterile glass vial for injectionMountainside Medical EquipmentAL-SV10020clear, sterile glass injection unit
20 gauge needleExel26417needles hypodermic 20g x 1" plastic hub (yellow) / regular bevel
22 gauge needleExel26411needles hypodermic 22g x 1" plastic hub (black) / regular bevel
deuterium oxideSigma-Aldrich151882-25G99.9 atom % D
isofluoraneVetone3060fluriso isoflurane, USP
OMNIC Spectra SoftwareThermoFisher Scientific833-036200FT-IR standard software
petroleum jellyVaseline305212311006Vaseline, 100% pure petroleum jelly, original, skin protectant
plastic capillary tubesInnovative Med Tech100050sodium heparin anticoagulant, 50 μL capacity, 30 mm length
Sealed liquid spectrophotometer SL-3 FTIR CAF2 CellInternational Crystal Laboratory0005D-8750.05 mm Pathlength
sodium chlorideEMD Millipore1.37017suitable for biopharmaceutical production
Thermo Electron Nicolet 380 FT-IR SpectrometerThermoFisher Scientific269-169400discontinued model, newer models available

References

  1. Schiffmann, C., Clauss, M., Hoby, S., Hatt, J. M. Visual body condition scoring in zoo animals – composite, algorithm and overview approaches. Journal of Zoo Aquarium Research. 5 (1), (2017).
  2. Peig, J., Green, A. J.

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