Evaluation of Vitamin A Status and Diagnosis of Hypovitaminosis A in Amphibians
American Association of Zoo Veterinarians Conference 2015
Allan P. Pessier1, DVM, DACVP; Carlos E. Rodriguez2, DVM, DACVP
1Wildlife Disease Laboratories, Institute for Conservation Research, San Diego Zoo Global, San Diego, CA, USA; 2Department of Animal Health, Disney’s Animal Kingdom, Orlando, FL, USA


Without supplementation, insect-based diets for amphibians and reptiles are known to be deficient in nutrients such as calcium and vitamin A.5 In the last decade, hypovitaminosis A has been recognized as a limiting factor for amphibian conservation programs that must successfully maintain and breed endangered species for ex situ rescue, survival assurance, and reintroduction programs.6,7 Empirical treatments, new approaches to dietary supplementation of insects (especially carotenoids and techniques for more effective gut-loading), and experimental studies have been useful for improving our understanding of this condition.2,4-6 However, research and clinical management have been hampered by practical considerations (e.g., it is difficult to get sufficient samples from very small frogs), limited basic information on vitamin A metabolism in amphibians, and a lack of standardization in diagnostic methods.3,7 A presumptive diagnosis of hypovitaminosis A in amphibians will often be made by a pathologist observing squamous metaplasia (SM) in a normally mucus-producing or ciliated epithelium.6,7 Although the tongue is the most common anatomic site for SM (i.e., short tongue syndrome), it is also recognized in the oropharynx, esophagus, ureter, reproductive tract, and cloaca. It is important to note that SM is not observed in every vitamin A-deficient animal, nor is it consistently observed in every anatomic site.7 Collection and histologic examination of a range of different tissues is suggested for every amphibian necropsy. Measurement of vitamin A levels in serum or liver is encouraged to confirm a diagnosis of hypovitaminosis A, but there are important pitfalls including awareness of what is measured and reported by different laboratories (retinol+retinyl esters or simply retinol) and the need for proper collection of samples (autolysis and light exposure influences results).3,7 Interpretation of vitamin A levels can be frustrating because of a lack of validated reference ranges in amphibians, wide variation in “normal” upper levels between individuals and species, and physiologic maintenance of serum retinol levels until deficiencies are advanced (serum retinol is not linear in relation to deficiency).1,3,7,8 However, low vitamin A levels (e.g., <5–10 µg/g retinol in liver) should always raise suspicion of deficiency.

Literature Cited

1.  Berkvens CN, Lentini A, Dutton CJ, Pearl DL, Barker IK, Crawshaw GJ. Serum and hepatic vitamin A levels in captive and wild marine toads (Bufo marinus). Zoo Biol. 2014;33:536–543.

2.  Brenes-Solo A, Dierenfeld ES. Effect of dietary carotenoids on vitamin A status and skin pigmentation in false tomato frogs (Dyscophus guineti). Zoo Biol. 2014;33:544–552.

3.  Clugston RD, Blaner WS. Vitamin A (retinoid) metabolism and actions: what we know and what we need to know about amphibians. Zoo Biol. 2014;33:527–535.

4.  Dugas MB, Yeager J, Richards-Zawacki CL. Carotenoid supplementation enhances reproductive success in captive strawberry poison frogs (Oophaga pumilio). Zoo Biol. 2013;32:655–658.

5.  Livingston S, Lavin SR, Sullivan K, Attard L, Valdes EV. Challenges with effective nutrient supplementation for amphibians: a review of cricket studies. Zoo Biol. 2014;33:565–576.

6.  Pessier AP. Short tongue syndrome and hypovitaminosis A. In: Mader DR, Divers SJ, eds. Current Therapy in Reptile Medicine and Surgery. Philadelphia, PA: Saunders; 2014:271–276.

7.  Rodriguez CE, Pessier AP. Pathologic changes associated with suspected hypovitaminosis A in amphibians under managed care. Zoo Biol. 2014;33:508–515.

8.  Sullivan KE, Fleming G, Terrell S, Smith D, Ridgeley F, Valdes EV. Vitamin A values of wild-caught Cuban tree frogs (Osteopilus septentrionalis) and marine toads (Rhinella marina) in whole body, liver, and serum. J Zoo Wildl Med. 2014;45:892–895.


Speaker Information
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Allan P. Pessier, DVM, DACVP
Wildlife Disease Laboratories
Institute for Conservation Research
San Diego Zoo Global
San Diego, CA, USA

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