Quantitating cantharidin levels in a blister beetle

August 14, 2024
Cat Barr | PhD, DACT, TVMDL Diagnostic Toxicologist Emeritus (retired)

In January, an animal owner in northwestern Washington State contacted the Texas A&M Veterinary Medical Diagnostic Laboratory (TVMDL) regarding possible blister beetles on their property. Specifically, the animal owner was interested in measuring the amount of cantharidin in a sampling of the beetles. Cantharidin is a very potent blistering agent, causing colic, heart arrhythmias and potentially death in horses. It is toxic to all mammalian species.

TVMDL had previously quantitated cantharidin in a different type of blister beetle1 and therefore received three individual beetles for testing from the animal owner in early March. A single large beetle (see photos) had been found on a barn wall, and a mating pair caught in the act on a fence. Based on the position when caught, the male was smaller, and since the female was of similar size to the beetle from the barn wall, they were both presumed to be female. Studies of blister beetle behavior and cantharidin content indicate the male generates cantharidin and transfers it to the female at mating2. The beetles were designated Female 1, Female 2 (from the pair), and Male and determined to be short-winged blister beetles (Meloe sp.).

Using the published method for quantification of cantharidin from beetles3, each beetle was dried for several days at approximately 60oC, then weighed before being crushed to a powder for extraction. To verify the extraction was effective, a sample of three-stripe blister beetles (Epicauta tembexa) was extracted and assayed alongside. Cantharidin was quantified against a solvent standard curve using gas chromatography/mass spectrometry (GC/MS). The following results were generated.

Sample IDCantharidin, % Dry Matter BasisCantharidin/ Beetle, mg# Beetles to Kill 1000 lb. Horse (1 mg/kg)
Meloe sp. Female 10.391.5303
Meloe sp. Female 20.160.5908
Meloe sp. Male0.891.3349
Epicauta temexa, mixed, 6 beetles1.430.222064

Three-stripe blister beetles are known to contain 1-12 % cantharidin on a dry matter basis4.

A much larger sampling would be needed to characterize this beetle further with regard to cantharidin content. The data suggests that short-winged blister beetles are probably less toxic than the three-stripe blister beetle. Their effect on animal and human communities is dependent on feeding and swarming habits.

The method TVMDL uses to test animal clinical specimens for cantharidin exposure is much more sensitive than the method employed on these beetles. Poisoned animals (usually horses on alfalfa) generally have cantharidin concentrations in serum, urine, or stomach/cecal content in parts per billion (ppb) levels when they are showing clinical signs. TVMDL uses liquid chromatography with a triple quadrupole mass spectrometer (LC/MSn) to achieve the sensitivity needed. 1.0 % = 10,000 parts per million (ppm); 1 ppm = 1000 ppb.

For more information about TVMDL’s test catalog, visit tvmdl.tamu.edu or call 1.888.646.5623.

REFERENCES

  1. Barr AC, et al. Cantharidin poisoning of emu chicks by ingestion of Pyrota insulataJ Vet Diagn Invest 10(1):77-79 (1998).
  2. Brown WD, Stanford R. Male mating tactics in a blister beetle (Coleoptera: Meloidae) vary with female quality. Can J Zoo 70(9):1652-1665 (1992).
  3. Carpinera JL, et al. Cantharidin levels in blister beetles (Coleoptera: Meloidae) associated with alfalfa in Colorado. J Econ Entomol 78:1052-1055 (1985).
  4. Ray AC, et al. Etiologic agents, incidence, and improved diagnostic methods of cantharidin toxicosis in horses. Am J Vet Res3(4):208-215 (1989).