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.

<b>四种马利筋蝶寄主（<i>Danaus</i>属）中新类簇虫原生动物的检出情况与宿主特异性</b>
宝拉·A·巴里加（Paola A. Barriga）^1*、埃莉诺·D·斯滕伯格（Eleanore D. Sternberg）^2,3、蒂埃里·勒弗夫尔（Thierry Lefèvre）^2,4、雅各布斯·C·德·罗德（Jacobus C. de Roode）²、索尼娅·阿尔蒂泽（Sonia Altizer）¹
¹ 美国佐治亚大学奥杜姆生态学学院，雅典，GA 30602，美国
² 埃默里大学生物学系，克利夫顿路1510号，亚特兰大，GA 30322，美国
³ 宾夕法尼亚州立大学传染病动力学中心，大学公园，PA 16802，美国
⁴ 法国国家科研中心-法国发展研究院MIVEGEC实验室（传染病与媒介：生态学、遗传学、进化与防控），阿格罗波利斯大道911号，34394蒙彼利埃，法国
*通讯作者：电子邮箱：[e-mail]

<b>元数据</b>
"完整调查"文件汇总了斑蝶亚科（Danainae）4种野生蝴蝶种群中卵囊簇虫（<i>Ophryocystis elektroscirrha</i>，简称OE）的感染率数据，涉及物种分别为君主斑蝶（<i>Danaus plexippus</i>）、女王斑蝶（<i>D. gilippus</i>）、热带女王斑蝶（<i>D. eresimus</i>）以及小漫游斑蝶（<i>D. petila</i>）。
本研究通过识别马利筋及其他对<em>Danaus</em>属蝴蝶具有吸引力的显花植物，在所述所有采样点内，于蝴蝶吸食花蜜或主动飞行时使用捕虫网进行采集。采用非破坏性方式采集蝴蝶腹部鳞片与寄生虫孢子以确定感染状态。具体方法参照Altizer等（2000）的描述：将直径2.5cm的透明胶带贴于斑蝶腹部，转移至索引卡片后计数所有孢子以确定孢子载荷量。
根据琥珀色色泽、椭圆形且两端渐尖的形态，以及10~14mm长、7~10mm宽的尺寸特征（Leong等，1997；Sander等，2013），将孢子样本鉴定为OE或类OE寄生虫。由于尚未通过分子分析验证君主斑蝶体内的OE寄生虫与女王斑蝶或其他寄主体内的寄生虫是否为同一物种，因此我们将君主斑蝶体内检出的寄生虫归类为OE寄生虫，而将其他蝶类体内检出的寄生虫归类为类OE寄生虫。
每支胶带样本的孢子数超过100个的个体被视为重度感染，此前的君主斑蝶实验表明，此类个体大概率在幼虫阶段摄入了孢子（Altizer等，2000）。数据集的感染列中，重度感染个体标记为"1"。反之，孢子数少于100个的情况可能源于成虫之间的被动孢子传播（De Roode等，2009、2007），此类个体被视为已暴露但未必感染，因此将其标记为"未感染"，对应数据库中的"0"值。

<b>数据列含义如下：</b>
Species：采集的蝴蝶物种
Year：蝴蝶采集年份
Population：蝴蝶采集地点
Infected：0 = 未感染，1= 感染

其余四个文件汇总了5项实验室条件下开展的实验结果，旨在检验OE或类OE寄生虫对君主斑蝶（<i>Danaus plexippus</i>）与女王斑蝶（<i>D. gilippus</i>）的感染特异性。具体而言，实验1与实验2以君主斑蝶寄生虫菌株分别侵染君主斑蝶与女王斑蝶寄主：实验1使用每头幼虫10个孢子的侵染剂量，实验2使用每头幼虫100个孢子的侵染剂量。实验3与实验4采用完全交互交叉侵染设计，分别使用从两种寄主体内采集的寄生虫侵染君主斑蝶与女王斑蝶。将实验3与实验4的结果合并以分析感染概率。针对实验3，我们进一步单独分析了成虫寿命（未合并数据，因实验4未测量寿命），对应文件标注为"实验3成虫寿命"。
实验5中，我们分别使用三种来源的寄生虫侵染来自两个种群（美国佐治亚州与澳大利亚昆士兰州）的君主斑蝶：三种寄生虫来源分别为北佛罗里达州的君主斑蝶寄生虫、澳大利亚的君主斑蝶寄生虫，以及澳大利亚小漫游斑蝶（<i>D. petila</i>）体内的类OE寄生虫，以此比较寄生虫相对于寄主物种与来源地的特异性。

<b>实验1至实验5以及实验3寿命文件的数据列含义如下：</b>
Host：研究的蝴蝶物种
Treatment：处理组，表明仅对经处理的个体进行分析
Infected：0 = 未感染，1= 感染
Dose：接种的寄生虫孢子数量
Year：实验开展年份
Monarchp：君主斑蝶的采样种群（美国佐治亚州或澳大利亚）
Parasite：寄生虫来源。在实验3与实验4中，该字段指代寄生虫采集自君主斑蝶还是女王斑蝶；在实验5中，该字段指代寄生虫采集自澳大利亚君主斑蝶、佛罗里达州君主斑蝶，还是"其他蝶类"（即澳大利亚<em>D. petila</em>体内的寄生虫）
Sex：蝴蝶性别，雄性（M）或雌性（F）
Life_span：蝴蝶存活天数。