DOI: 10.5061/dryad.d51c5b05b
title: Data from: Phormia regina allele report

description: Genetic structure of blow fly (Diptera: Calliphoridae) populations has
 remained elusive regardless of high relatedness within wild-caught
 samples. The aims of this research were to determine if the implementation
 of a high-resolution spatiotemporal sampling design would reveal latent
 genetic structure among blow fly populations and to elucidate
 environmental impacts on structure. Adult female black blow flies, Phormia
 regina (Meigen), were collected from nine urban parks in Indiana, USA over
 three years. Fly gut DNA was genotyped at six microsatellite loci, with
 subsequent amplification and sequencing of vertebrate mitochondrial DNA
 from the same source. Flies were also screened for vertebrate fecal
 metabolites. Latent clustering revealed four genetic groups which were
 interpreted as 11 distinct temporal populations. An analysis of molecular
 variance of temporal populations revealed stronger genetic differentiation
 (FST = 0.048, F’ST = 0.664) relative to geographic populations (FST =
 0.009, F’ST = 0.241). Mean kinship within temporal populations was higher
 than expected in a panmictic population (R = 0.032 ± 0.088). Weather
 conditions (i.e., wind speed, precipitation, humidity, temperature) and
 vertebrate resource availability in the local environment significantly
 impacted the observed genetics of P. regina. Twenty-five vertebrate
 species were detected from flies, and 16% of flies collected in 2016 –
 2017 tested positive for vertebrate feces, suggesting many varied
 resources are important for maintaining high gene flow among geographic
 populations. A complex interplay between biotic and abiotic factors, as
 well as the flies’ own extensive dispersal abilities, seems to drive the
 strong temporal structure of this species in the Midwestern US.
description: <i>Blow Fly Collections</i> Spatiotemporal blow fly collections were made over a three-year period (2015 – 2017) in Central Indiana, USA. Blow flies were sampled from nine geographic sites over 36 sampling periods. Sampling periods occurred from June to September in 2015 and from March to October in 2016 and 2017. Collection sites consisted of urban parks and other public areas in four different counties (Table 1).  Fly collections were made with an aerial sweep net and an aged chicken liver bait inside a mesh-screened container to prevent flies from physically landing on the bait without hindering their ability to detect the smell of the bait [86]. Fly sampling lasted 20 min starting at the time of bait exposure. Temperature and relative humidity were measured throughout the collections using a Datalogging RH/Temperature Pen (SPER Scientific, Scottsdale, Arizona, USA) elevated ~1 m above the ground. Wind speed and direction were also measured at three arbitrary times during each collection using a Digi-Sense® Mini Vane Anemometer (Cole-Parmer, Vernon Hills, Illinois, USA). After the 20 min collection window, flies were immediately killed in 70% ethanol for transportation to the lab and stored at -20˚C. Flies were identified to species using a dichotomous key [87] and fly richness and evenness per sample were calculated using the <i>vegan </i>package [88] in R Studio v1.4.1717 [89]. <i>Dissections and DNA Extractions</i> Gut dissections from adults of the black blow fly, <i>Phormia regina</i> (Meigen), were performed according to previously published protocols [90] on a maximum of 10 randomly chosen females from each sample. A standard phenol:chloroform:isoamyl alcohol DNA extraction method was performed on each individual gut dissection, with a final elution in 35 uL of TE buffer. This procedure allowed for the simultaneous extraction of fly DNA (from the gut epithelial tissue) and vertebrate DNA (from the last meal within the fly gut). The normally discarded phenol layer of the DNA extraction was also screened for fecal metabolites [90]. <i>Genotyping</i> Six polymorphic microsatellite loci were used to assess population structure [24]. Amplification of each sample was carried out in two separate multiplex reactions each containing three primer pairs. One reaction contained primers L3, L9, and L12, and the other contained primers L8, L13, and L14. Each reaction consisted of 5.0 μL 2x PCR MasterMix (Promega), 0.5 μL 1x Bovine Serum Albumin (BSA), 0.5 μL nuclease-free water, 1 μL of each primer, and ~1ng DNA. Concentrations for each primer were as follows: 2.5 μM L3, 1.5 μM L9, 5.0 μM L12, 2.5 μM L8, 2.5 μM L13, and 2.5 μM L14. PCR conditions consisted of an initial 2 min denaturation step at 95˚C, nine touchdown cycles of a 15 s at 95˚C, 30 s from 63˚C to 55˚C, and 72˚C for 2 min, followed by 22 cycles of 95˚C for 15 s, 55˚C for 30 s, and 72˚C for 2 min, with a final extension at 72˚C for 7 min (Mastercycler Pro thermocycler, Eppendorf®, Hamburg, Germany). Following amplification, 1 μL of each PCR product was added to 9.7 μL HiDi formamide (Thermo Fisher Scientific, Waltham, MA) and 0.3 μL LIZ internal size standard (Thermo Fisher Scientific) for fragment analysis on a 3500 genetic analyzer (Thermo Fisher Scientific) and alleles were scored using GeneMarker® software (SoftGenetics®, State College, PA, USA).
description: ## Phormia regina allele report https://doi.org/10.5061/dryad.d51c5b05b
 This dataset contains the alleles for 1198 individual blow flies of the
 species *Phormia regina*, collected from nine urban parks in Indiana from
 2015 to 2017. There are six microsatellite loci tested across 159
 spatiotemporal populations, which have been grouped into 13 genetic
 clusters. **Description of the data and file structure** Below are
 descriptions of each acronym and variable found in the dataset. Ind =
 individual fly Temporal Population (1 through 11) ST Pop = spatiotemporal
 population (1 through 159) Pop = "Site" (see descriptions below)
 + "Sampling timepoint" (variable by year) + \_"Year"
 (15, 16, 17) MP = Military Park (Indianapolis) SP = Skiles-Test Park
 (Indianapolis) BR = Broad Ripple Park (Indianapolis) WL = Westlake Church
 (Indianapolis) LA = Purdue University (West Lafayette) BL = Upper Cascades
 Park (Bloomington) NW = Northwest Park (Indianapolis) UP = University Park
 (Greenwood) PP = Province Park (Franklin) Other columns: Microsatellite
 Loci (L12, L3, L13, L9, L8, L14) Scores for each allele are in two
 separate columns Failure to amplify an allele = 0 **Sharing/Access
 information** N/A **Code/Software** N/A