DOI: 10.5061/dryad.0cfxpnw3f
title: Influence of habitat quality and resource density on breeding-season female monarch butterfly (Danaus plexippus) movement and space use in north-central USA agroecosystem landscapes

description:  The eastern North American monarch butterfly is at risk of
 quasi-extinction due, in part, to the loss of breeding habitat in
 agricultural landscapes of the USA Midwest. Because adult females are not
 patch residents, egg abundance and distribution across the landscape are a
 function of their perceptual range, flight directionality, and flight step
 lengths. Conservation actions that account for habitat use in agricultural
 landscapes can enhance functional connectivity. Field-captured females
 (n=114) were released in a 64-ha area containing restored prairies,
 grass-dominated fields, and crop fields in Floyd County, Iowa, USA, and
 two 1,000 m linear north-south sections of grass-dominated roadside along
 secondary roads (~ 35 ha) with different proximity to prairie habitat in
 rural Story County, Iowa. Radio-tagged or untagged monarchs were released
 in areas with high-density, low-density, and zero density of forage and
 oviposition resources, as well as on habitat edges between high and
 zero-density habitats. Monarchs were observed for one hour. Radio-tagged
 individuals that flew beyond visual detection were relocated using
 handheld radio telemetry. Monarchs moved within and between habitat
 classes and typically performed up-wind search behavior. Monarchs
 successfully located resources, with some flying over 500 m to find
 high-density areas, providing evidence that the monarch's perceptual
 distance is &gt; 100 m. Regardless of habitat class or field site,
 most step lengths were &lt; 50 m, and turn angles were directional.
 Large steps (≥ 50 m) crossing habitat boundaries occurred with
 approximately half of the monarchs, which may indicate initiation of
 long-range searches for suitable habitat, consistent with their vagile
 behavior. Establishing habitat patches 50 m apart in agricultural
 landscapes would facilitate efficient movement. This study provides an
 extensive dataset of directly observed breeding-season monarch butterflies
 to assess the utilization of agricultural landscapes. Documentation of
 step lengths &gt; 50 m in complex, agricultural landscapes would not
 have been possible without the aid of radio telemetry. Results provide
 improved estimates of perceptual range and flight patterns within and
 between habitat patches that support models that simulate natural
 population dynamics to enable conservation planning at a landscape scale.
description: <i>Field Sites </i>            
 Three field sites were selected that contained areas with varying
 densities of milkweed ramets and blooming inflorescences in different
 habitat classes (see Table 1 for a glossary of key terms). The selected
 sites did not have structural boundaries (e.g., forest edges, see Schultz
 et al. 2012). A detailed description of each site is provided in S1-S4.
 The mosaic site was a 64-ha<sup> </sup>area in Floyd County,
 Iowa that contained a restored prairie (HD), grass-dominated fields (LD),
 and crop fields (ZD) (Fig 1A). The roadsides were two north-south, 1,000 m
 by 350 m linear sections (~ 35 ha) of grass-dominated roadside (LD) along
 secondary gravel roads in Story County, Iowa, with different proximity to
 prairie habitat. One roadside was bordered by maize to the west (ZD) and a
 remnant prairie (HD) to the east (RHD; Fig 1B, green box). The other
 roadside was bordered by maize and soybean fields to the east and west
 (ZD) and was ≥ 500 m from prairie habitat (RZD; Fig 1B, blue box).
 Observations occurred from June to August at the mosaic field site in 2019
 and the roadside field sites in 2020.  
 <i>Tracking Field-Captured Female Monarch Butterflies
 </i>            
 Field-captured, female monarchs with minimally worn wings were held for
 1-4 days and maintained as previously reported (Fisher et al. 2020, Fisher
 and Bradbury 2021). Monarch movement was observed on days with a
 temperature of 18.6-35.4°C and wind speed below 15.0 kph; wind direction
 was recorded prior to the release of each monarch. Monarchs were
 anesthetized with CO<sub>2</sub> and radio-tagged with a 220
 mg LB-2X transmitter (~ half the average weight of a monarch; Holohil
 Systems Ltd, Ontario, Canada; see Fisher et al. 2020). Untagged
 individuals served as controls. Monarchs were released on<i> Bromus
 inermis </i>or <i>Zea mays </i>within a habitat class
 (Table 1). Those that did not move from their release location (S5) within
 15 minutes were considered unresponsive, and the observation period was
 terminated (see Zalucki and Kitching, 1984). There was no significant
 difference in responsiveness between tagged and untagged individuals
 (S6-S8), consistent with previous studies that documented no impact on
 flight ability (Fisher et al. 2020, Fisher and Bradbury 2021).
 Consequently, observations of responsive tagged and untagged monarchs were
 combined for data analyses, resulting in 18-20 responsive individuals for
 each habitat class (Table 1).             Monarchs were
 observed visually by technicians stationed within a field site (yellow,
 green, and blue boxes in Fig 1). Locations between natural flight steps
 were georeferenced (Trimble Geo7x; Sunnyvale, CA). The amounts of time and
 behaviors performed (i.e., foraging, ovipositing, resting, mating) at each
 location and the amount of time spent flying between locations were
 recorded. In cases where monarchs flew beyond the limit of visual
 detection (ca. 50 m), monarchs were relocated with the aid of handheld
 radio telemetry equipment (directional 3-element Yagi antennae
 [164.000–166.000 MHz; Johnson's Telemetry, El Dorado Springs, MO],
 connected to VHF receivers [Alinco DJ-X11; Toyama, Japan] with 0.9-m coax
 cables). We also employed an automated radio telemetry system (Fisher et
 al. 2021) at the mosaic site; however, location estimates from the
 automated system were unsuccessful (S9). Monarchs that flew within the
 initial 15 min, but then remained stationary for 30 min, were recaptured.
 For the remaining responsive monarchs, capture was attempted after an
 observation period of 3,600-sec (one hour) within a field site. When a
 monarch was relocated and captured with the aid of handheld radio
 telemetry outside of a field site, the elapsed time from release to
 recapture was recorded. Unsuccessful relocation efforts were terminated
 three hours post-release. Consequently, observation periods ranged from 75
 – 7,500 s (1.25 – 125 min).              
 <i>Measurements</i>             Space use estimates with
 continuous-time movement models (ctmm) were created for each monarch using
 the ctmm package (Calabrese et al. 2016, Fleming et al. 2016, Fleming and
 Calabrese 2020) in R (R Core Team 2020) with the Ornstein-Uhlenbeck
 Foraging (OUF) process (see S10 for methodological details). These models
 produced 0.05 m<sup>2</sup> raster cells with occurrence
 probabilities ranging from 0-1. Straight-line measurements were used to
 analyze movement paths, assuming direct movement between georeferenced
 locations where monarchs were observed between flights. Step lengths
 between two consecutive georeferenced locations, bearing orientation of
 steps, and turn angles created from three consecutive georeferenced
 locations within a habitat class were calculated using the 'movement.
 pathmetrics' package in Geospatial Modeling Environment (GME; Spatial
 Ecology LLC, www.spatialecology.com/gme/) in combination with ArcMap 10.3. Total distances traveled were calculated by summing step lengths for each monarch. The Euclidian distances and bearings of Euclidian movement were calculated from the release locations to the location at the end of the observation period using ArcMap 10.3.     <i>Statistical Analyses</i>             All statistical analyses associated with movement characteristics were conducted in R (R Core Team 2020). The total and Euclidian distance measures for each individual were compared with a paired t-test. Distances traveled, step lengths within and between habitat classes, logit transformed proportions (Warton and Hui 2011) of raster cells with non-zero occurrence probability, as well as the time and georeferenced locations in each habitat class, were analyzed by field site and release habitat class with generalized linear models in the package 'emmeans' (Lenth et al. 2020). The Rayleigh test of uniformity, the Watson-Williams test for homogeneity of means, and the Watson-Wheeler test for homogeneity of angles with the 'circular' package (Lund 2017) were used to analyze orientations of movement relative to wind direction and distributions of turn angles within habitat classes. The percent of steps ≥ 50 m that originated in each habitat class were analyzed with multiple chi-square tests of independence and a Bonferroni corrected significance threshold (see Sedgwick 2012 for multiple comparisons).