title: Skunk musk causes methemoglobin and Heinz body formation in vitro.
abstract: BACKGROUND
  A captive Red Panda developed a regenerative anemia with Heinz bodies after being sprayed by a skunk. A definite cause-and-effect relationship between skunk musk and oxidative erythrocyte damage has not been reported, but it was suspected in one reported case of a dog with Heinz body hemolytic anemia.
  
  
  OBJECTIVE
  The objective was to determine whether skunk musk induces oxidative HGB damage in vitro.
  
  
  METHODS
  Plasma and RBC were harvested from heparinized blood of 3 dogs, 3 cats, and a Red Panda. Skunk musk was solubilized in ethanol and mixed with plasma from each species to make stock solutions of 4% musk and 4% ethanol. Aliquots of RBC were resuspended in autologous stock solutions and solvent controls to yield musk concentrations of 0%, 0.04%, and 0.4% (by volume). Aliquots were incubated at 37°C for 4-72 hours and assessed for oxidative damage by visual inspection, optical absorbance spectroscopy, transmission electron microscopy, and light microscopy after Wright and vital New Methylene Blue staining.
  
  
  RESULTS
  Dose-dependent brown color and absorption changes characteristic of methemoglobin were present by 4 hours and increased over 24 hours (Red Panda) and 72 hours (dog and cat). Similarly, there were time-dependent (all species) and dose-dependent (dog and cat) increases in the number of Heinz bodies, which were present by 4 hours and numerous by 24 hours.
  
  
  CONCLUSIONS
  In vitro, skunk musk causes Heinz body and methemoglobin formation in canine, feline, and Red Panda RBC, supporting the clinical association between Heinz body hemolytic anemia and skunk spray exposure.
authors.affiliation: Pathobiology and Diagnostic Investigation, Michigan State University, East Lansing, MI, USA.
authors.forename: Brittney R
authors.initials: BR
authors.lastname: Fierro
authors.forename: Dalen W
authors.initials: DW
authors.lastname: Agnew
authors.forename: Ann E
authors.initials: AE
authors.lastname: Duncan
authors.forename: Andreas F
authors.initials: AF
authors.lastname: Lehner
authors.forename: Michael A
authors.initials: MA
authors.lastname: Scott
mesh_terms: D053822:Ailuridae; D000745:Anemia, Hemolytic, Congenital / Q000097:blood / Q000209:etiology / Q000473:pathology / Q000662:veterinary*; D000818:Animals; D002415:Cats; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000473:pathology; D005229:Fatty Acids, Monounsaturated / Q000009:adverse effects*; D005260:Female; D006366:Heinz Bodies / Q000473:pathology; D012889:Mephitidae; D008706:Methemoglobin / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission / Q000662:veterinary; D018384:Oxidative Stress
chemical_list: D005229:Fatty Acids, Monounsaturated; C008563:musk; D008706:Methemoglobin
keywords: Anemia; RBC; oxidative damage