Status of Disease in the Emperor Penguins of Auster Rookery

Metadata record for data from ASAC Project 2954.See the link below for public details on this project.Public The primary goal of this project is to determine the status and origin of diseases in Emperor Penguins at Auster Rookery near Mawson Station, Antarctica. We will investigate the origins of such disease and the role humans may have played. We will sample adults and chicks in order to isolate and describe the pathogens. A high percentage of Emperor Penguin chicks have antibodies to infectious bursal disease (IBDV). This study will investigate the role the adults play in transmitting IBDV to their chicks. The high prevalence of IBDV antibodies should help us to isolate the virus and discover its origin.Project objectives:Status of Disease in the Emperor Penguins of Auster Rookery1) To determine the prevalence of disease in adult Emperor Penguins (Aptenodytes forsteri) at Auster Rookery. Over 65% of Emperor Penguin chicks at Auster Rookery had serum antibodies to Infectious Bursal Disease Virus (IBDV) in December 1995 (Gardner et al. 1997). We have no information on the presence of the same antibodies on adult Emperor Penguins. We will focus on IBDV, but will also test for other common avian diseases.2) To repeat the sampling of Emperor Penguin Chicks by Gardner et al. (1997) in order to compare the prevalence of IBDV in Emperor Penguin chicks in 2008 with 1995.3) To determine the seasonal progression of IBDV antibody prevalence in both adults and chicks.4) To determine the possible source(s) of viral infection in Emperor Penguin chicks. Because Gardner et al. (1997) had no information on adult Emperor Penguins we do not know how the chicks are exposed to IBDV. Gardner et al. (1997) suggested that poultry waste from the nearby Mawson Station may be a source of virus for the Emperor Penguin chicks. Penguins do not scavenge food, however, so the source of infection must be either from the environment, local predators/scavenger, or from parents feeding their young. By sampling both adults and chicks in different parts of the season, we will determine when antibodies first appear in the chicks. If they have antibodies in the early season, then scavengers/predators can not account for their exposure to IBDV.5) To determine the source of the IBDV. We will attempt to isolate virus from Emperor Penguins in order to identify the strains responsible for the antibody reactions in Emperor Penguins. Using reverse transcription polymerase chain reaction (RT-PCR), we will sequence genes from the virus which can be compared with known gene sequences from serotypes available from GenBank.6) To monitor the chick mortality and conduct field necropsies of the Emperor Penguins at the Auster colony to determine whether IBDV or other diseases are a factor in reproductive success.7) To contribute to a conservation strategy for Antarctic wildlife. By developing information on the importance and origins of disease in Emperor Penguins we can clarify the role of human visitors in the transmission of disease in Antarctic wildlife. Progress against objectives:Excellent progress has been made towards all the stated goals.By spending the winter of 2008 at Mawson Station we had access to the emperor penguin colony at Auster. We successfully sampled 400 adults and 200 chicks as stated in the proposal. We now have the samples back in Australia and analyses are beginning. No sample analysis for disease could be undertaken while still in Antarctica.The samples will 1) give us a determination of the prevalence of IBDV antibodies (also some other disease viruses) for both adults and chicks. 2) One of our sampling periods was a repeat of the sampling conducted by Gardner et al. so we can compare the prevalence of IBDV antibodies in chicks from 2 different years and relate that to the prevalence in adults. 4) We conducted our sampling at four different times during the winter so that we were able to sample chicks before any other species visited the colony, then sample them again 6 weeks after skuas and giant petrels were in the area. 5) In order to determine the source of IBDV we want to determine its RNA sequence. To that end, a full set of samples have been sent to Dr. Daral Jackwood at Ohio State University, a colleague and IBDV expert. He has just begun to analyse the samples to isolate and sequence the IBDV RNA. 6) We monitored chick mortality with visits to the colony on average once per week. We noted approximately 800 dead chicks and collected 120 of the chicks for field necropsies. They mostly died of starvation with a few exceptions. We found parasites in one dead chick. We also collected 9 carcasses of adult emperor penguins. Eight of the 9 were females who all died with complications of egg laying. 7) This goal will require the completion of all the analyses for us to make conservation recommendations.This collection of files represents the data (including samples) collected for Project 2954 on Emperor Penguins.This project was based on collecting samples from Emperor penguins throughout the winter season. There were 5 sets of samples.1-Adults during courtship in May2-Adults during hatch period in early August3-Chicks at ca 5 weeks of age--soon after leaving the constant care of their parents4- Adults in early summer (mid-November)5-Chicks in early summer (late November)Sampling at these intervals it was hoped we would be able to discriminate when or how the IBD antibodies appear in this population.The files are:1) Project 2954-Emperor IBD Antibodies PrevalenceSummaryThis is a summary file of all the Virus Neutralization tests to determine the presence of antibodies to Infectious Bursal Disease (IBD) in the 5 groups of Emperor Penguins.2) Project 2954-Emperor IBD summary graphThis is a graphical representation of the summary information for IBDV antibodies3) Project 2954-Emperor Penguin Egg Samples-2008This is a listing of the abandoned eggs collected over the winter. The eggs were weighed and measured. Many were sampled. We took 5ml of yolk and stored the samples in -80 C for future analysis.4) Project 2954-Emperor Penguin sampling2008This is the master file of sampling. Each penguin we handled was given a sample identification (but no permanent identifying marks). Each was weighed, measured, inspected for ticks and samples taken. This file identifies which samples were obtained. In addition it represents some analyses. The sexes of many of the penguins have been determined genetically with PCR. That is included in the file.5) Project 2954-Emperor Penguin Serology NDV and AI 2008This file is a listing of all the serum samples and the results of Hemagglutination Inhibition test (HAI) for antibodies to Newcastle Disease Virus (NDV) and an antibody ELISA test for Avian Influenza. No sign of any exposure to NDV or AI in Emperor penguins.6) Project 2954-Disease Status Poster-Peng Conf 2010This Powerpoint file is a single oversized page that presents much of the results that we have to date. It was presented at the 7th International Conference on the Biology of Penguins, in Boston USA, Sept 20107) Project 2954-Searching for IBDV in Emperors with RT-PCRThis MS WORD file is the summary of methods and results of testing tissue samples from chick carcasses found at the colony. Bursa and spleen tissue was preserved and sent to Daral Jackwood in Ohio USA. He conducted real time Reverse Transcriptase PCR on the samples to try to find the IBD virus. That is essential for the full identification of the source of antibodies in these penguins. Unfortunately all results were all negative. We hope with a new proposal to expand our testing to find this virus.

元数据记录（Metadata record）对应ASAC 2954号项目的数据集。可通过下方链接查看该项目的公开详情。本项目的核心目标为探明南极莫森站附近奥斯特企鹅聚居地帝企鹅的疾病现状与传染源。我们将探究此类疾病的起源，以及人类活动可能起到的作用。我们将对成年企鹅与幼雏进行采样，以分离并描述病原体。1995年12月的调查显示，奥斯特聚居地超过65%的帝企鹅幼雏携带传染性法氏囊病病毒（Infectious Bursal Disease Virus, IBDV）抗体，本研究将探究成年企鹅向幼雏传播IBDV的具体路径。高流行的IBDV抗体水平将助力我们分离病毒并明确其起源。

项目目标：
1. 探明奥斯特聚居地成年帝企鹅（Aptenodytes forsteri）的疾病流行情况。此前我们尚无该聚居地成年帝企鹅相关抗体的监测数据。本次研究将以IBDV为核心关注对象，同时也会检测其他常见禽类传染病。
2. 重复Gardner等人（1997）的幼雏采样工作，以对比2008年与1995年帝企鹅幼雏的IBDV流行率。
3. 明确成年企鹅与幼雏体内IBDV抗体流行率的季节变化规律。
4. 探明帝企鹅幼雏病毒感染的潜在来源。由于Gardner等人（1997）未涉及成年帝企鹅的相关数据，我们尚不清楚幼雏的IBDV暴露途径。该团队曾提出，附近莫森站的家禽废弃物可能是幼雏感染病毒的来源，但帝企鹅不会食腐，因此感染来源只能是环境、本地捕食者/食腐动物，或是亲代育雏时的传播。通过在不同季节节点对成年企鹅与幼雏进行采样，我们将明确幼雏体内抗体首次出现的时间。若幼雏在季初便已携带抗体，则可排除食腐/捕食动物作为暴露途径的可能。
5. 明确IBDV的传染源。我们将尝试从帝企鹅体内分离病毒，以鉴定引发帝企鹅抗体反应的病毒毒株。我们将通过逆转录聚合酶链反应（reverse transcription polymerase chain reaction, RT-PCR）对病毒基因进行测序，并与GenBank中已公开的血清型基因序列进行比对。
6. 监测幼雏死亡率，并对奥斯特聚居地的帝企鹅进行野外剖检，以明确IBDV或其他疾病是否会影响繁殖成功率。
7. 为南极野生动物保护策略提供支撑。通过明确帝企鹅疾病的重要性与起源，我们可以厘清人类访客在南极野生动物疾病传播中所扮演的角色。

目标推进情况：目前所有既定目标均已取得良好进展。2008年我们在莫森站越冬，得以进入奥斯特帝企鹅聚居地。我们按照研究计划成功采集了400只成年企鹅与200只幼雏的样本。目前样本已运回澳大利亚，分析工作即将启动。在南极境内时，我们暂未开展疾病相关的样本检测工作。
样本分析将达成以下目标：1) 明确成年企鹅与幼雏体内IBDV抗体（及部分其他病毒）的流行率；2) 我们的其中一个采样周期与Gardner等人的采样方案一致，因此可对比两年间幼雏的IBDV抗体流行率，并结合成年企鹅的流行情况进行关联分析；4) 我们在冬季开展了四次不同时段的采样，得以在其他物种到访聚居地前采集幼雏样本，随后在贼鸥与巨海燕活跃于该区域的6周后再次采样；5) 为明确IBDV的来源，我们需要确定其RNA序列。为此，全套样本已寄往美国俄亥俄州立大学的达里尔·杰克伍德（Daral Jackwood）博士——他是IBDV领域的专家，也是我们的同行。他已开始分析样本，以分离并测序IBDV RNA；6) 我们通过平均每周一次的聚居地走访监测幼雏死亡率，共记录到约800只死亡幼雏，并收集了120只幼雏进行野外剖检。大部分幼雏死于饥饿，仅少数存在其他死因；我们在1只死亡幼雏体内发现了寄生虫。此外我们还收集了9具成年帝企鹅的尸体，其中8只为雌性，均死于产卵并发症；7) 该目标需待所有分析工作完成后，我们才能提出针对性的保护建议。

本文件集合包含了2954号项目中帝企鹅相关的所有采集数据（含样本信息）。本项目在整个冬季周期内采集了帝企鹅样本，共分为5组：
1. 5月繁殖期的成年企鹅
2. 8月初孵化期的成年企鹅
3. 约5周龄的幼雏——即脱离亲代持续照料后不久的个体
4. 夏初（11月中旬）的成年企鹅
5. 夏初（11月下旬）的幼雏
我们期望通过该间隔采样方案，明确该种群中IBD抗体出现的时间与传播路径。

文件清单如下：
1) Project 2954-Emperor IBD Antibodies PrevalenceSummary：本文件为所有病毒中和试验的汇总结果，用于检测5组帝企鹅体内传染性法氏囊病（IBD）抗体的存在情况。
2) Project 2954-Emperor IBD summary graph：本文件为IBDV抗体汇总信息的可视化图表。
3) Project 2954-Emperor Penguin Egg Samples-2008：本文件为冬季采集的废弃企鹅卵的清单。所有卵均进行了称重与测量，其中多数被采样：我们提取了5ml卵黄，并将样本保存于-80℃以备后续分析。
4) Project 2954-Emperor Penguin sampling2008：本文件为采样总记录。每只被捕捉的企鹅均被赋予样本编号（但未添加永久标识），并进行了称重、测量、蜱虫检查及样本采集。本文件记录了所有获取的样本信息，同时包含部分分析结果：我们通过聚合酶链反应（Polymerase Chain Reaction, PCR）对多数企鹅的性别进行了遗传鉴定，相关信息已纳入本文件。
5) Project 2954-Emperor Penguin Serology NDV and AI 2008：本文件为所有血清样本的清单，以及新城疫病毒（Newcastle Disease Virus, NDV）血凝抑制试验（Hemagglutination Inhibition test, HAI）和禽流感（Avian Influenza, AI）抗体酶联免疫吸附试验（ELISA）检测的结果。结果显示，帝企鹅未表现出NDV或AI感染的迹象。
6) Project 2954-Disease Status Poster-Peng Conf 2010：本Microsoft PowerPoint文件为单页超大尺寸展板，展示了我们目前已获得的大部分研究成果，该展板于2010年9月在美国波士顿举办的第7届国际企鹅生物学大会上展示。
7) Project 2954-Searching for IBDV in Emperors with RT-PCR：本Microsoft Word文件为聚居地内发现的幼雏尸体组织样本检测方法与结果的汇总。我们保存了法氏囊与脾脏组织，并寄往美国俄亥俄州的达里尔·杰克伍德处。他对样本进行了实时逆转录PCR检测，以寻找IBD病毒，但所有结果均为阴性。我们希望通过新的研究提案扩大检测范围，以分离出该病毒。