Usually means: Substance or method lethal to mules.
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We found 7 dictionaries that define the word mule killer:

General (6 matching dictionaries)
  1. mule killer: Merriam-Webster
  2. mule-killer, mule killer: Wordnik
  3. mule killer: Wiktionary
  4. Mule killer: Dictionary.com
  5. mule killer: FreeDictionary.org
  6. Mule killer: TheFreeDictionary.com

Computing (1 matching dictionary)
  1. mule killer: Encyclopedia

Definitions from Wiktionary (mule killer)

noun:  (US, Southern US) Any of various unrelated arthropods, including whip scorpions, velvet ants, mantises, wheel bugs, and walking stick insects, that have been erroneously believed to kill livestock by stinging them or being swallowed.

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