Name: reduced itraconazole concentration and durations are successful in treating batrachochytrium dendrobatidis infection in amphibians
Uploaded: 2014-03-14T18:45:00
Description: Watch this Scientific Journal Video about Reduced Itraconazole Concentration and Durations Are Successful in Treating Batrachochytrium dendrobatidis Infection in Amphibians at JoVE.com

我要感谢CL理查兹-Zawacki帮助概念化的实验设计和协助材料，A. Pessier帮助制定治疗方案和尸检分析，MWH查特菲尔德，MJ Robak，K.塔斯克和D Lenger在帮助实验室和支持和M.麦克法登和P.哈洛在Taronga动物园的拍摄过程中使用其空间。该项目是通过授予洛杉矶Brannelly路易斯安那州环境教育委员会授予和生命技术部分资金由野生动物和渔业的路易斯安那部门。

伦理批准：本研究及其方法已获美国杜兰大学动物使用及护理委员会（协议没有0407）。 1。测试用于 BD感染<ol><li>擦拭用于 BD <ol><li>拭动物用无菌棉签5倍的ventrum，每个大腿上，每一侧，并且每个肢为总共45笔画五倍。擦拭取样是对波特的两栖类皮肤无创性方法。 </li><li>在与招之间轻轻转动棉签，保证DNA的最大量收集棉签。 </li><li>于1.5ml微?...

<ol>
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使用此协议，我们能够安全，有效地降低两栖动物伊曲康唑治疗波特的治疗剂量和时间。最低治疗剂量和时间（六天0.0025％）是一次成功的治疗亚临床感染一nebulifer。这两个较低的剂量和治疗时间的效果表明，有必要为临床治疗试验的波特感染的治疗，因为目前用于两栖类的所有物种和年龄阶层没有安全有效的治疗方法。 死亡由伊曲康唑治疗造成的原因?...

两栖类动物正在经历的所有脊椎动物类群1的生物多样性的最大跌幅。壶菌病，引起壶真菌病原体Batrachochytrium壶菌（BD）一种皮肤疾病，是这些戏剧性的下滑2的主要原因之一波特是最糟糕的病原体上记录造成生物多样性丧失。它感染了600种两栖动物和有造成下跌超过300种，其中有多达200种已成为极度濒危或灭绝2,3。两栖动物壶菌是不特定于主机的，目前尚不清楚什么样的素质的主机允许波特感染进步导致严重的疾病。壶菌病引起的死亡通过破坏离子通道的皮肤，造成电解质损失和心脏衰竭4。此外， 屋宇署上清液（不含波特游动孢子），导致蝌蚪的免疫反应5 leadin克研究者认为波特产生的副产品6有毒。 由于许多物种正在迅速下降，俘虏保证殖民地和育种计划是重要的保护措施，并可能对某些物种的唯一生命线。建立和维持无病殖民地，是两栖动物管理设施是一个挑战。目前的化疗方法治疗波特感染，虽然有效，可能会损害正在接受治疗的动物，并没有单一的治疗方法一直是跨物种和两栖类7龄级成功。 对波特感染治疗几个临床试验已经完成7。治疗的三种最常用的方法有热疗法，氯霉素和伊曲康唑。热疗是，似乎有一些副作用，可以在多个生命阶段中使用的nonchemotherapeutic治疗选择，并是根儿盟友有效。在治疗试验中，查特菲尔德和Richards-Zawacki 8发现，治疗动物的96％被安置在30℃下10天之后清除了感染。虽然这次审判是不是100％成功的，它提供给别人评价热疗法为他们自己种的基准。热疗法，虽然有效的许多物种，是不切实际的，可以不保持在高温环境中长时间许多高山物种。氯霉素，抗菌，已经成功地用于治疗波特在许多物种中，并在体外抑制真菌的生长。作为抗菌，氯霉素不攻击真菌特别，但已经知道导致细胞凋亡在真核细胞9。据推测，氯霉素治疗引起细胞凋亡的皮肤，造成细胞内的孢子囊，但确切的作用机制尚不清楚。虽然没有临床试验已经进行，因此，剂量和治疗时间没有被评估，当前的治疗方案通常需要浸没为两至四个星期10,11，使得这种治疗不切实际的地面两栖动物。氯霉素也被限制在一些国家的兽医使用，因为人体暴露已经与再生障碍性贫血12一种罕见形式的关联。 伊曲康唑，唑类抗真菌，是目前最广泛地用于在圈养两栖动物治疗波特感染。最广​​泛使用的协议是入浴动物每天连续11天五分钟0.01％伊曲康唑溶液。这个协议是来自用于哺乳动物施用的标准，并且，同时有效两栖类，它不基于在两栖动物11,13的临床试验。这个协议可以导致某些物种的负面影响，特别是在属蛙和蝌蚪和最近metamorphs 11。蝌蚪治疗已经成功使用伊曲康唑的浓度低得多（0.0005％），但用色素脱失14相关联。这同样浓度造模上最近metamorphs，结果为不成功在治疗感染15。因为有一个负面影响与伊曲康唑治疗的风险，一些从业者已经使用一半的通常浓度（0.005％），连续11天，发现这是成功的固化波特感染16。 本研究是使用伊曲康唑治疗波特感染15临床治疗试验的总结。该协议描述，试图找到最低剂量和治疗时间，既安全，有效的治疗波特感染动物与伊曲康唑。在这项研究中使用的动物是最近的墨西哥湾沿岸蟾蜍metamorphs，Incilius nebulifer。

<table border="0" cellpadding="0" cellspacing="0" width="503">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:179px;">0.1% Aqueous Itraconazole, Sporonox</td>
			<td style="width:147px;">Janssen Pharmaceutica Inc.</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:179px;">NaCl</td>
			<td style="width:147px;">Fisher Scientific</td>
			<td style="width:177px;">S640-500</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:179px;">KCl</td>
			<td style="width:147px;">Fisher Scientific</td>
			<td style="width:177px;">BP366-1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:179px;">CaCl2</td>
			<td style="width:147px;">Fisher Scientific</td>
			<td style="width:177px;">BP510-100</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:179px;">NaHCO3</td>
			<td style="width:147px;">Fisher Scientific</td>
			<td style="width:177px;">BP328-500</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:179px;">Sterile cotton swabs</td>
			<td style="width:147px;">Medical and Wire Equipment</td>
			<td style="width:177px;">MW113</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:179px;">1.5 ml micro tubes</td>
			<td style="width:147px;">Sarstedt</td>
			<td style="width:177px;">73.703.217</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:179px;">Qiagen DNEasy Blood and Tissue Kit</td>
			<td style="width:147px;">Qiagen</td>
			<td style="width:177px;">69581</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:179px;">7500 Real-time PCR</td>
			<td style="width:147px;">Applied Biosystems</td>
			<td style="width:177px;">4351104</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:179px;">VIC IPC</td>
			<td style="width:147px;">Applied Biosystems</td>
			<td style="width:177px;">4308323</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:179px;">BSA</td>
			<td style="width:147px;">Applied Biosystems</td>
			<td style="width:177px;">AM2618</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:179px;">Agar</td>
			<td style="width:147px;">Invitrogen</td>
			<td style="width:177px;">30391023</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:179px;">Tryptone</td>
			<td style="width:147px;">Invitrogen</td>
			<td style="width:177px;">Q10029</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:179px;">Penicillin-streptomycin</td>
			<td style="width:147px;">Invitrogen</td>
			<td style="width:177px;">15070-063</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:179px;">Petri Dish</td>
			<td style="width:147px;">Sigma Aldrich</td>
			<td style="width:177px;">CLS430589</td>
		</tr>
	</tbody>
</table>

reduced itraconazole concentration and durations are successful in treating batrachochytrium dendrobatidis infection in amphibians

两栖动物遇到任何脊椎动物类的最大跌幅和这些下降的主要原因是真菌病原体，Batrachochytrium壶菌（BD），从而导致疾病的壶菌病。圈养保证菌落受威胁的两栖动物物种的重要全球范围内，可为那些面临灭绝的关键威胁的唯一生命线。维持无病殖民地，是圈养管理者优先级，为所有物种和整个生命阶段还没有安全有效的治疗方法尚未确定。最广​​泛使用的化学治疗是伊曲康唑，尽管通常使用的剂量可能是有害的一些个体和物种。我们进行了临床治疗试验，以评估伊曲康唑的低，但更安全有效剂量是否能找到治愈波特感染。我们发现，从0.01-0.0025％，减少了治疗浓度，减少治疗时间从115分钟浴-6日，青蛙可以用较少的副作用，减少治疗相关死亡率治愈波特感染。

伊曲康唑治疗被认为是成功的，如果所有的动物都是BD-阴性治疗结束后四个星期。治疗前开始，接种后两周内，所有的BD-接种动物受到感染，以1,121.7平均游动孢子负荷（±17.34），而治疗结束四周后，屋宇署阳性对照组动物有16,765.12平均游动孢子负荷（ ±14643.08）。 Incilius nebulifer的治疗组（n = 15），成功利用浴场0.0025％，伊曲康唑浓度为五分钟，每...

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Reduced Itraconazole Concentration and Durations Are Successful in Treating Batrachochytrium dendrobatidis Infection in Amphibians

Batrachochytrium壶菌感染两栖动物可以使用缩小浓度（0.0025％）和伊曲康唑的持续时间较短（5分钟浴六天），比一般用被淘汰。采用这种改性的治疗方案更少的死亡率和较少副作用进行观察。

减少伊曲康唑的浓度和持续时间是成功的治疗<em&gt; Batrachochytrium壶菌</em&gt;感染两栖动物

Amphibians are experiencing the greatest decline of any vertebrate class and a leading cause of these declines is a fungal pathogen, Batrachochytrium ...

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Reduced Itraconazole Concentration and Durations Are Successful in Treating Batrachochytrium dendrobatidis Infection in Amphibians

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Use of Artificial Sputum Medium to Test Antibiotic Efficacy Against Pseudomonas aeruginosa in Conditions More Relevant to the Cystic Fibrosis Lung

Use of Artificial Sputum Medium to Test Antibiotic Efficacy Against Pseudomonas aeruginosa in Conditions More Relevant to the Cystic Fibrosis Lung

使用人工痰介质测试抗生素药效试验<em&gt;绿脓杆菌</em&gt;更多相关的肺囊肿性纤维化的条件

There  is growing concern about the relevance of in vitro antimicrobial  susceptibility tests when applied to isolates of P. aeruginosa from  cystic ...

Culturing Microglia from the Neonatal and Adult Central Nervous System

培养小胶质细胞的新生和成年的中枢神经系统

Microglia are the resident macrophage-like cells of the central nervous system (CNS) and, as such, have critically important roles in physiological and ...

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

总蛋白提取和二维凝胶电泳方法<em&gt;鼻疽</em&gt;物种

The investigation of  the intracellular protein levels of bacterial species is of importance to  understanding the pathogenic mechanisms of diseases ...

Neutrophil Isolation and Analysis to Determine their Role in Lymphoma Cell Sensitivity to Therapeutic Agents

中性粒细胞分离和分析，以确定他们的淋巴瘤细胞的敏感性对治疗药物的作用

Neutrophils are the most abundant (40% to 75%) type of white blood cells and among the first inflammatory cells to migrate towards the site of ...

Isolation of Salmonella typhimurium-containing Phagosomes from Macrophages

Isolation of Salmonella typhimurium-containing Phagosomes from Macrophages

巨噬细胞中含有吞噬的鼠伤寒沙门氏菌的分离

Salmonella typhimurium is a facultative intracellular bacterium that causes gastroenteritis in humans. After invasion of the lamina propria, S. ...

Analysis of Iophenoxic Acid Analogues in Small Indian Mongoose (Herpestes Auropunctatus) Sera for Use as an Oral Rabies Vaccination Biological Marker

Analysis of Iophenoxic Acid Analogues in Small Indian Mongoose Herpestes Auropunctatus Sera for Use as an Oral Rabies Vaccination Biological Marker

小印度蒙古狼的异苯抗酸模拟分析(赫皮斯特奥罗普塔图斯)塞拉,用作口服狂犬病疫苗接种生物标记

The small Indian mongoose (Herpestes auropunctatus) is a reservoir of rabies virus (RABV) in Puerto Rico and comprises over 70% of animal rabies cases ...

Recurrent Escherichia coli Urinary Tract Infection Triggered by Gardnerella vaginalis Bladder Exposure in Mice

Recurrent Escherichia coli Urinary Tract Infection Triggered by Gardnerella vaginalis Bladder Exposure in Mice

由加德纳菌阴道膀胱暴露在小鼠中引发的复发性大肠杆菌尿路感染

Recurrent urinary tract infections (rUTI) caused by uropathogenic Escherichia coli (UPEC) are common and costly. Previous articles describing models of ...

Separation of Immune Cell Subpopulations in Peripheral Blood Samples from Children with Infectious Mononucleosis

传染性单核细胞增多症儿童外周血样本中免疫细胞亚群的分离

Infectious mononucleosis (IM) is an acute syndrome mostly associated with primary Epstein&#8211;Barr virus (EBV) infection. The main clinical symptoms ...