Occurrence and host specificity of a neogregarine protozoan in four milkweed butterfly hosts (Danaus spp.)

<b>Occurrence and host specificity of a neogregarine protozoan in four milkweed butterfly hosts (<i>Danaus</i> spp.)</b> Paola A. Barriga^1*, Eleanore D. Sternberg^2,3 Thierry Lefèvre^{2, 4}, Jacobus C. de Roode², Sonia Altizer¹ ¹ Odum School of Ecology, University of Georgia, Athens, GA 30602, USA ² Biology Department, Emory University, 1510 Clifton road, Atlanta, GA 30322, USA ³ Center for Infectious Disease Dynamics, Pennsylvania State University, University Park, PA 16802, USA ⁴ MIVEGEC lab (Maladies Infectieuses et Vecteurs: Ecologie, Génétique, Evolution et Contrôle), CNRS-IRD, 911, Av. Agropolis 34394 Montpellier France *Corresponding author: E-mail address: [e-mail] <b>Metadata</b> The “complete survey” file compiles information about prevalence of <i>Ophryocystis elektroscirrha</i> (OE) infection in wild populations of four butterfly species in the family Danainae (<i>Danaus plexippus</i>,<i> D. gilippus</i>,<i> D. eresimus</i>,<i> and D. petila</i>). To collect these butterflies, milkweed and other flowering plants attractive to <i>Danaus</i> spp. were identified and butterflies were collected using an aerial net during nectar feeding or active flight in all the locations mentioned. Butterfly abdomens were sampled non-destructively to obtain scales and parasite spores to determine infection status. Spore load was determined by counting all of the spores in a 2.5 cm diameter transparent adhesive tape pressed against monarch abdomens and transferred to index cards as described in Altizer <i>et al.</i> (2000). Spore samples were identified as OE or OE-like parasites based on amber coloration and shaped as ovals with tapered ends, and, with dimensions of 10-14 mm length and 7-10 mm width (Leong et al., 1997; Sander et al., 2013). Since molecular analyses have not yet been performed to test whether OE parasites found on monarchs belong to the same species as those found on queens or other species, we categorized parasites as OE parasites when found on monarchs, and as OE-like parasites when found on other butterfly species. Samples with more than 100 spores per adhesive tape were considered heavily infected, as previous monarch experiments demonstrated that these butterflies likely ingested spores as larvae (Altizer et al., 2000). Heavily infected butterflies were marked in the infected column of the dataset as “1.” In contrast, spore loads of less than 100 spores can result from passive spore transfer between adult butterflies (De Roode et al., 2009, 2007), and we refer to butterflies with these lower numbers of spores as exposed, but not necessarily infected. Therefore, those samples were marked as no infected and represent the “0s” in the database. <b>Data columns represent:</b> Species: Butterfly species collected Year: Year when butterflies were collected Population: Location where the butterflies were collected Infected: 0 = no infected and 1= infected The other four files compile the results of five experiments performed in laboratory conditions to test the specificity of OE or OE-like parasites infection in <i>D. plexippus</i> (monarchs) and <i>D. gilippus</i> (queens). Specifically, Experiments 1 and 2 focused on monarch and queen hosts, challenging each species with monarch parasite strains; the first experiment used a parasite dose of 10 spores and the second experiment used a dose of 100 spores per larva. Experiments 3 and 4 focused on a fully reciprocal cross-infection design challenging monarchs and queens with parasites collected from each of the two host species. Results from Experiment 3 and 4 were pooled to analyze infection probability. For experiment 3, we further analyzed adult monarch lifespan (without pooling the data, as lifespan was not measured for experiment 4). This file is labeled as “Experiment 3 adult life span.” In Experiment 5, we challenged monarchs from each of two populations (Georgia/U.S. and Queensland/Australia) with parasites from each of three sources: monarch parasites from North Florida, monarch parasites from Australia and an OE-like parasite from the lesser wanderer (<i>D.</i> <i>petilia</i>) in Australia, to compare the specificity of parasites in relation to host species and source location. <b> </b> <b>Data columns represent (on files Experiment 1 to 5, and Experiment 3 life span):</b> Host: Butterfly species studied Treatment: Treated to emphasize that only treated butterflies were analyzed Infection: 0 = no infected and 1= infected Dose: Number of parasite spores inoculated Year: When experiments were performed Monarchp: Explains the population where monarchs were collected (Georgia, US, or Australia). Parasite: Origin of the parasite. In Experiments 3 and 4 it refers to if those were collected from monarch or queen butterflies. In Experiment 5 it refers to if collected from monarchs in Australia, Florida or from “Other butterfly” and in that case comes from <i>Danaus petilia</i>. Sex: whether the butterfly was male (M) or female (F) Life_span: number of days that butterflies lived.