Study 2: How does sampling methodology (swab vs feces) affect
recovered community composition?
To assess the efficacy of cloacal swab and fecal sampling in S.
virgatus , we collected 8 adult lizards (n = 4 females and n = 4 males)
on 30 May 2019 using the same sites and methodology described above.
Animals were housed individually in sterile 15 X 23 cm plastic tanks
lined with paper towels that had also been sterilized with 70% ethanol
on a west-facing screened porch of SWRS’s Live Animal Holding Facility.
They were offered a single cricket on the day of capture that was
removed if not eaten by ~1200 the next day. Sterile
water was provided ad libitum , and one heat lamp on a 14:10 light
cycle was shared between two adjacent tanks.
Lizard cloacae were swabbed first at ~0500 on 31 May,
before animals had woken up and defecated; this is considered our
pre-defecation swab sample. Beginning at 0600, the tanks were checked
for feces every 30 min, until 1830, when the heat lamps had been off for
30 min and the lizards were no longer active. When a fecal pellet was
found, it was collected with sterile tweezers and the lizard’s cloaca
was immediately swabbed. Based on this method, all post-defecation swabs
were taken within 30 min of the defecation event. We also collected a
control swab, which we used to sample the researchers’ hands, a lizard’s
external vent and belly, and the air of the porch, to collect any
microbes that may have contaminated cloacal swabs during sampling.