Filth flies have long been implicated in pathogen transmission routes in which it is thought that they acquire pathogens directly from infected feces and mechanically transmit them to humans and animals. Currently, however, no analytical method for identifying feces as the source of fly-related pathogens exists for applied settings.  A qualitative method to detect various tetrapyrrole urobilinoids associated with vertebrate feces was created in order to definitively link flies to fecal resources. Flies from three feeding treatments (unfed flies, liver fed flies, and feces fed flies; N = 20 flies/treatment) were analyzed using a Thermo LTQ-XL mass spectrometer (San Jose, CA) coupled to an Agilent 1100 HPLC.  The sample was separated using reversed phase chromatography on a 100 x 2.1 mm C18 column at a flow rate of 200 uL/min. The solvents were 0.1% formic acid in water (solvent A) and 0.1% formic acid in 70:30 acetonitrile:methanol (solvent B). The separation began with an initial 1 minute hold at 30% B followed by a 9 minute linear gradient from 30 to 95% B.  Data were acquired in positive ion mode.  Two peaks indicating the presence of the compound class of interest were present at a retention time of ~6.5 minutes containing the m/z 343 and ~ 8.2 minutes containing the m/z 466. Confirmatory tests were run on fly extracts where different types of animal feces were consumed to verify that the urobilinoid compounds would be present regardless of dietary restrictions.<br/>

LC-MS/MS detects urobilinoids from feces in fly guts

255th National Meeting (2018)

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