作者感谢云南，中华人民共和国和阿博维尔，科特迪瓦，他们参与上述研究的社区。我们非常感激的测试和执行的协议，协助当地的野队。我们特别感谢下列同事从瑞士热带和公共健康研究所:Endriss伊薇特提供在视频拍摄过程中的显微图像，Hanspeter实验室的支持和孙燕姿克劳思·马蒂和他的团队对她的帮助。最后，我们感谢两位匿名审稿进行了一系列有益的意见。

1。改良加藤技术<ol><li>将一个标准的改良加藤模板为41.7毫克（mg），大便显微镜载片上有一个洞。 </li><li>炒到2-3克（g）的新鲜粪便样品放在一张报纸或铝箔，然后按顶部的金属丝或塑料网筛它一块。 </li><li>使用一个小的塑料小铲，刮离网筛分的材料，并完全填充改良加藤模板的孔中。以除去过量的粪便物，平与刮刀的孔的含量。 </li><li>垂直删除，而不会干扰现在坚持粪便的显微镜幻灯片模板。模板和抹刀可以在水中用洗涤剂清洗，并重复使用。 </li><li>将一块玻璃纸，预浸泡在甘油孔雀石绿溶液在至少24小时（小时），在显微镜载玻片上的粪便样品。 </li><li>要传播的粪便成厚厚的涂抹，轻轻按下一个干净的微样品滑动的滑动范围，均匀分布的材料内的圆的直径略小于显微镜载玻片的广度。 </li><li>允许的幻灯片，以清除为30-60分钟（min），在此期间，幻灯片必须远离阳光直射的地方。然后，系统地研究了厚厚的涂抹在光学显微镜下（40到100倍的放大倍率）。数一数......鸡蛋和分层物种（即，A.lumbricoides，T.trichiura，和钩虫）。其他寄生虫卵（如血吸虫 ）值得注意的是，也可以通过该方法来检测，并应分开计数和记录。 </li><li>要得到感染的强度的一个标准化的估计，通常表示为每克粪便（EPG）的卵数，乘以蛋计数由24（24×41.7毫克≈1克）。 </li></ol> 2。贝尔曼技术<ol><li>建立一个独立，可容纳一个玻璃漏斗和连接橡胶管，关闭机智公顷剪辑，漏斗底部。 </li><li>形状象一个圆锥体的金属丝网筛的筛或一块放置在液漏斗和层与医用纱布一块。 </li><li>填补了漏斗用自来水，并确保在整个设置也不会漏出。 </li><li>舀到中间的医用纱布20-30克新鲜粪便，并确保它是完全浸没在水中。如果需要添加更多的水。医用纱布折叠过的样品。 </li><li>从下面，人造光照耀在2小时的漏斗。 </li><li>漏到离心管中的50毫升（ml）的下部，在漏斗中的水轻轻释放的橡胶管上的剪辑。 </li><li>在600×g离心5分钟（相对离心力）离心收集的水，并小心地倒出上清液，而不会干扰沉积物。保留最后几滴在管。 </li><li>重悬沉淀中加入2滴在显微镜载片上的解决方案和检查幻灯片在光学显微镜下确认的物种进行检测和100-400X的放大倍率（40倍放大）。第一阶段的幼虫（L1）的S。粪类圆线虫是活的，积极的姿态，在显微镜下。 </li></ol> 3。 20米折返跑测试<ol><li>测量一个20米的平直运行过程，并把它标记为锥。 </li><li>帮助测试参与者佩戴一个心脏监测仪上的手腕和相应的发射器周围的胸部18。此设备允许参与者的心脏速率试验之前和之后进行测量，从而监控该参与者是否已经产生了极大的努力。 </li><li>指导参与者在球场上来回运行，通过预先录制的声音信号，划定20米圈的步伐。从8.5千米每小时（公里/小时）的行驶速度，频率的信号指示一个相当于0.5公里/小时每分钟20米的间隔增加。 </li><li>确保日在参与者开始一个新的20米的间隔，每次一个信号响起，下一个信号到达之前完成。 </li><li>停止参与者，如果他/她没有跟随的步伐，连续两个20米的间隔。 </li><li>记录的最高运行速度参与者完成充分，沿着与该参与者的年龄和性别，最后的间隔表示。 </li><li>验证，在测试结束的心脏率达到是孩子的最大心脏率19。如果没有达到这个心脏率，孩子可能不会产生了最大的努力，因此，应重复测试。 </li><li>估计参与者的最大有氧能力（VO最多2个 ），使用Léger的和他的同事提出的一个方程:最大VO 2 = 31.025 + 3.238 *速度（公里/小时） - 3.248 *年龄（岁）+ 0.1536 *速度（公里/小时）*年龄（岁）。 </li></ol> 4。代表性的成果在运输及房屋局局长诊断与改良加藤技术，独特的特点，不同的寄生虫卵允许的识别大多数物种20。的蛋A.lumbricoides（约45-75微米（μm）长，宽和35-50微米， 图1），比T.trichiura（ 图2），钩虫（ 图3）的蛋具有较厚的壳壁。一个受精卵A.lumbricoides（ 图1）具有外胚乳的外衣，这使得它看起来呈金黄色，而一个未受精的卵子缺乏这方面的外层披毛和稍微细长，更大的整体。鸡蛋T.trichiura（约50μm的长，宽和22微米， 图2）中是细长的，并具有不同的极性的插头。钩虫鸡蛋（约64-76微米的长，宽和36-40微米， 图3），在另一方面，有一个非常薄的壁，随后由明确的环围绕的致密的细胞群在中心的蛋。 s.stercoralis·L-1幼虫（ 图4）是约180-380微米长，并有一个很短的口腔内。一个明显的生殖器原基（突出显示一个蓝色的箭头在图4）可以看出，和幼虫（突出显示一个红色箭头， 图4）的尾部有一个尖底。 作为真正的影响STH感染儿童的体能仍存在争议13,14,15，有代表性的结果，在这一点上他们的影响。可以用各种方式进行20米折返跑测试结果的解释。可以比较谁是相似的年龄，性别，其他可能相关的健康状况，社会经济地位和文化习惯的感染和非感染科，儿童的体能。这可以通过在一个横截面的调查13,14,15运输及房屋局局长感染状况及感染者和非感染儿童的身体素质进行评估在一个特定的时间点。证据的强度会更高时，以下的适龄儿童的体能评估之前和之后驱虫/安慰剂治疗的随机对照试验设计，使短期和长期的评估。可以进一步尝试找出一个尽可能小的剂量-反应关系，运输及房屋局局长感染的强度和体能与体能，并比较不同程度的multiparasitism 1。在工作邑和他的同事15，强身健体，最大VO 2所描述的，被认为是显着减少感染与T.trichiura的儿童相比，孩子们谁没有被感染与T.trichiura（ 图5） 。 <img alt="图1" src="/files/ftp_upload/3966/3966fig1.jpg" /> Figu<html>重1。显微图像的A.lumbricoides鸡蛋受精（A）和（B）500倍的放大倍率下未受精的。比例尺= 50微米。 <img alt="图2" src="/files/ftp_upload/3966/3966fig2.jpg" /> 图2。500倍的放大倍率下的T.trichiura蛋。比例尺=50μm的显微图像。 <img alt="图3" src="/files/ftp_upload/3966/3966fig3.jpg" /> 图3的钩虫鸡蛋500倍的放大倍率下的显微图像。比例尺= 50微米。 <img alt="图4" src="/files/ftp_upload/3966/3966fig4.jpg" /> 图4的S.stercoralis·L-1幼虫在250倍放大倍数下的显微图像。箭头指示的尾巴（红色）和生殖器原基（蓝色）。比例尺= 100微米。 d/3966/3966fig5.jpg"/&gt; 图表显示的体能相比，的儿童感染T.trichiura水平的体能水平的儿童没有感染T. 图5。 虫 。

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没有利益冲突的声明。

本文中所描述的三个协议已经过测试，并执行在西南部云南省的一部分，中华人民共和国各成员之间的布朗族少数民族21在不同地区的非洲6,13,14。 有几个模板，可用于改良加藤技术。每一个不同的孔尺寸将提供一个独特的粪便量。对于计算的EPG，因此，有必要知道单元的模板，以乘以鸡蛋的计数量的修正因子，以到达的EPG估计。此外，当在社区钩虫是本调查?...

注意:在此列表中的材料是通用的，可以从不同的来源获得。 <table border="1"><tbody><tr><td> 的试剂的名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>改良加藤试剂盒 [400带有一个孔的6毫米（mm）（直径）在1.5毫米厚的模板塑料模板400塑料抹刀; 80网目尺寸的尼龙筛20米的作用; 20米的作用的亲水性的玻璃纸，34微米厚。] </td><td>韦斯特弗兰德森</td><td></td><td>请注意的甘油孔雀石绿溶液不包含在该试剂盒中，但可以从任何化学品供应商购买。 </td></tr><tr><td>玻璃漏斗[约8厘米（cm），宽6厘米深] </td><td></td><td></td><td>为贝尔曼技术在玻璃漏斗应该是能够容纳约60毫升水。钍E尺寸的橡胶软管和纱布是依赖于所使用的玻璃漏斗的类型。 </td></tr><tr><td>生物显微镜</td><td>奥林巴斯</td><td> CX21LED/CX21 </td><td></td></tr><tr><td>离心分离</td><td>上海医疗器械集团</td><td> 80 - 2T </td><td></td></tr><tr><td>预先录制的信号</td><td> Bitworks设计</td><td>团队哔哔测试版1.3.1 </td><td> iPhone应用程序</td></tr><tr><td>心脏监测仪</td><td> POLAR </td><td> FT1 </td><td>手表和发射器</td></tr></tbody></table>

determining soil-transmitted helminth infection status and physical fitness of school-aged children

土壤传播的蠕虫（STH）感染是常见的。事实上，超过1亿人受到影响，主要是在发展中世界，那里的贫困盛行，卫生行为，供水和卫生设施往往缺乏1,2， 蛔虫 ， 鞭虫和钩虫种类， 十二指肠钩虫 ， 钩虫美洲 ，是最普遍的STHs 3。由于钩虫病，蛔虫病和鞭虫病的全球疾病负担的估计是22.1，10.5，和640万伤残调整寿命年（DALY），分别为4。此外，估计有30-100亿人感染粪类圆线虫 ，最被忽视的STH种具有全球意义，可以说是也造成相当大的公众健康的影响5,6。多物种感染（即不同的STHs窝藏在一个单一的个体）是常见的，已经感染林<html>糟透了降低生产率，从而促进经济的发展中国家的前景1,3。 的诊断的常见STHs的，世界卫生组织（WHO）建议的改良加藤技术7.8，这是一个相对简单的方法来确定这类感染的发生率和强度。它促进了检测，感染者通过在其粪便中的寄生虫卵。 随着的诊断S.stercoralis方面，目前还没有简单而准确的工具。的的贝尔曼技术是应用最广泛的方法，其诊断。的贝尔曼技术背后的原理是，积极S.stercoralis幼虫迁移出照明的新鲜粪便样品的幼虫趋光9。它需要不太复杂的实验材料和消耗更少的时间比文化和免疫学方法5。 T"发病率与运输及房屋局局长感染范围从急性，但共同的症状，如腹痛，腹泻，皮肤瘙痒，慢性症状，如贫血，不足和营养不良，和认知功能障碍10。一般都是非特异性的症状，微妙的，他们经常被忽视，被认为是一个正常的条件下受影响的个体，或者被处理作为可能会对多见于一个给定的设定的其他疾病的症状，因此，可以想象的是，真正的负担被低估了评估的STH感染工具依靠自我宣称的症状和体征通常情况下人口为基础的调查。 在20世纪80年代末和90年代初，Stephenson和他的同事们强调STH感染的可能性降低了体能的男孩6-12岁11,12。最近13,14,15这条线的科学调查获得了新的动力。 20米（m）折返跑测试开发ð验证莱热等。，是全球通用的衡量有氧健身的儿童17。测试是简单的规范和可以执行的地方有20米长，平坦的运行过程和音频源，使得它的使用有吸引力的资源限制设置13。为方便和规范的尝试，在评估是否STH感染有效果的适龄儿童的体能，我们提出的方法，诊断STH感染或措施的体能是简单的执行，提供精确和可重复的结果。这将有助于产生新的证据，STH感染对健康的影响。

Watch this Scientific Journal Video about Determining Soil-transmitted Helminth Infection Status and Physical Fitness of School-aged Children at JoVE.com

Determining Soil-transmitted Helminth Infection Status and Physical Fitness of School-aged Children

用土壤传播的的蠕虫（STHs）的慢性感染引起吸收不良，发育迟缓，浪费在成长中的孩子。因此，它是合理的，这些感染也可以减少儿童的体能。在这里，我们可视化技术的诊断STHs和20米折返跑测试，以评估儿童的身体健康。

确定土壤传播的蠕虫感染状况和体质的学龄儿童

Soil-transmitted helminth (STH) infections are common. Indeed, more than 1 billion people are affected, mainly in the developing world where poverty ...

determining-soil-transmitted-helminth-infection-status-physical

Universidad de Cartagena UDEC

Research

JoVE Journal

Immunology and Infection

23.3K 次观看.  Swiss Tropical and Public Health Institute, Basel, Switzerland. 用土壤传播的的蠕虫（STHs）的慢性感染引起吸收不良，发育迟缓，浪费在成长中的孩子。因此，它是合理的，这些感染也可以减少儿童的体能。在这里，我们可视化技术的诊断STHs和20米折返跑测试，以评估儿童的身体健康。

视频: Determining Soil-transmitted Helminth Infection Status and Physical Fitness of School-aged Children

Measuring the Functional Abilities of Children Aged 3-6 Years Old with Observational Methods and Computer Tools

The analysis of functional abilities and their development in early childhood (0-6 years old) are fundamental aspects among young children with certain types of developmental difficulties that can facilitate prevention, through programmed interventions adapted to the needs of each user (student or patient). There are, however, few investigations to date, that have analyzed the use of automated tools for recording and interpreting the results of the initial assessment. Here, a protocol is presented to examine the functional abilities in early childhood in young children, aged between 3-6 years old, with intellectual disabilities, but the protocol can also be used for ages 0 to 6 years. The protocol makes use of a computer application, eEarlyCare, that facilitates the interpretation of the results of systematic observations, which are recorded in natural environments by professionals trained in early intervention. The software can be used to analyze 11 functional areas (Food Autonomy, Personal Care and Hygiene, Dressing and Undressing Independently, Sphincter Control, Functional Mobility, Communication and Language, Daily Life Routines, Adaptive Behavior and Attention) and a total of 114 different behaviors. Its use facilitates the analysis of the observed abilities and greatly assists early intervention. Compared to other observational methods, it allows a more efficient use of personal and material resources. The use of the computer application facilitates the recording of the observation results, which helps with organization and reflection on the observations. The software displays the observation results on-screen compared to normal developmental parameters. This information can be referred to for decision-making about the most suitable intervention program for each user (student or patient). Likewise, clustering techniques are applied to analyze the relation between the type of intellectual disabilities and functional development identified with the software, a relation that is intended to serve as a guide for early-care professional intervention.

We present a protocol to use a computational tool to record and analyze the functional abilities of children aged 3-6 years old. The protocol facilitates the comparison of these abilities throughout their development and can be used to assess developmental difficulties.

用观察方法和计算机工具测量3-6岁儿童的功能能力

The analysis of functional abilities and their development in early childhood (0-6 years old) are fundamental aspects among young children with certain ...

科研

行为学

Breakfast Habits among Schoolchildren in the City of Uruguaiana, Brazil

Breakfast is the first and most important meal of the day, and omitting it is associated with a greater probability of overweight and school absenteeism. The eating habits constituted in children depend on social, educational and economic factors, and they usually perpetuate into adulthood. For this reason, the most propitious stage for health promotion interventions on healthy nutrition is childhood. Specifically, in Brazil, malnutrition is relevant in children from low-income families although, currently, there are few studies that relate nutrition with the social factors. In this study, a reliability analysis of the Eating&#160;Habits of the School&#160;Population questionnaire was conducted, and the items referring to the breakfast habits and to some sociodemographic variables were selected. Subsequently, a total of 470&#160;fourth&#160;grade pupils self-completed the questionnaire in 12&#160;schools in Uruguaiana&#160;(Brazil). More than 50% of the mothers and more than 70% of the fathers worked in diverse areas of services and sales in stores or supermarkets. With regards to the hypothesis contrast, a significant relation was observed between the frequency of having breakfast with the father or the mother and having had breakfast&#160;(p&#160;&#60;&#160;0.001). This type of study favors a population analysis for the subsequent design and execution of health promotion activities.

This article describes the breakfast habits and the factors related to schoolchildren in a city of Brazil&#160;(Uruguaiana). To this end, a transcultural adaptation was performed of a questionnaire validated in Spain, and 470&#160;boys and girls from twelve&#160;schools were interviewed.

巴西乌鲁瓜亚纳市学童的早餐习惯

Breakfast is the first and most important meal of the day, and omitting it is associated with a greater probability of overweight and school absenteeism. ...

Physical Activity Measurement in Children Accepting Table Tennis Training

An increasing body of evidence now shows that the majority of children in China experience lower levels of physical activity (PA) than the recommended guideline. Table tennis is a compound and technically difficult game that is popular in China; undertaking table tennis training in clubs can help children to elevate their levels of PA. Given that children cannot complete self-evaluated questionnaires themselves and caregiver-based observations are not suitable for children, we hypothesized that an actigraphy-based method can be an objective method to measure PA. In the present study, we describe a procedure that can be used to evaluate PA levels using an actigraphic device and software. Furthermore, since hip-worn devices are known to reduce compliance, we attempted to assess the agreement between hip-worn and wrist-worn device data. Collectively, our results indicate that these devices are suitable for measuring PA and leisure time physical activity (LTPA) levels. Together with subjective questionnaires, both hip-worn and wrist-worn devices are highly suitable for evaluating PA in Chinese children undergoing table tennis training in clubs.

This study proposes an accelerometer-based method to objectively measure physical activity (PA) and leisure time physical activity (LTPA) in Chinese children accepting table tennis training in clubs.

接受乒乓球训练的儿童的身体活动测量

An increasing body of evidence now shows that the majority of children in China experience lower levels of physical activity (PA) than the recommended ...

Developing a Salivary Antibody Multiplex Immunoassay to Measure Human Exposure to Environmental Pathogens

The etiology and impacts of human exposure to environmental pathogens are of major concern worldwide and, thus, the ability to assess exposure and infections using cost effective, high-throughput approaches would be indispensable. This manuscript describes the development and analysis of a bead-based multiplex immunoassay capable of measuring the presence of antibodies in human saliva to multiple pathogens simultaneously. Saliva is particularly attractive in this application because it is noninvasive, cheaper and easier to collect than serum. Antigens from environmental pathogens were coupled to carboxylated microspheres (beads) and used to measure antibodies in very small volumes of human saliva samples using a bead-based, solution-phase assay. Beads were coupled with antigens from Campylobacter jejuni, Helicobacter pylori, Toxoplasma gondii, noroviruses (G I.1 and G II.4) and hepatitis A virus. To ensure that the antigens were sufficiently coupled to the beads, coupling was confirmed using species-specific, animal-derived primary capture antibodies, followed by incubation with biotinylated anti-species secondary detection antibodies and streptavidin-R-phycoerythrin reporter (SAPE). As a control to measure non-specific binding, one bead set was treated identically to the others except it was not coupled to any antigen. The antigen-coupled and control beads were then incubated with prospectively-collected human saliva samples, measured on a high throughput analyzer based on the principles of flow cytometry, and the presence of antibodies to each antigen was measured in Median Fluorescence Intensity units (MFI). This multiplex immunoassay has a number of advantages, including more data with less sample; reduced costs and labor; and the ability to customize the assay to many targets of interest. Results indicate that the salivary multiplex immunoassay may be capable of identifying previous exposures and infections, which can be especially useful in surveillance studies involving large human populations.

In the current climate of scarce resources, new technologies are emerging that allow researchers to conduct studies cheaper, faster and with more precision. Here we describe the development of a bead-based salivary antibody multiplex immunoassay to measure human exposure to multiple environmental pathogens simultaneously.

制定唾液抗体多重免疫测定来测量人体暴露于环境中的病原体

The etiology and impacts of human exposure to environmental pathogens are of major concern worldwide and, thus, the ability to assess exposure and ...

免疫与感染

Sexual Transmission of American Trypanosomes from Males and Females to Naive Mates

American trypanosomiasis is transmitted to humans by triatomine bugs through the ingestion of contaminated food, by blood transfusions or accidently in hospitals and research laboratories. In addition, the Trypanosoma cruzi infection is transmitted congenitally from a&#160;chagasic mother to her offspring, but the male partner&#39;s contribution to in utero contamination is unknown. The findings of nests and clumps of amastigotes and of trypomastigotes in the theca cells of the ovary, in the goniablasts and in the lumen of seminiferous tubules suggest that T. cruzi infections are sexually transmitted. The research protocol herein presents the results of a family study population showing parasite nuclear DNA in the diploid blood mononuclear cells and in the haploid gametes of human subjects. Thus, three independent biological samples collected one year apart confirmed that T. cruzi infections were sexually transmitted to progeny. Interestingly, the specific T. cruzi antibody was absent in the majority of family progeny that bore immune tolerance to the parasite antigen. Immune tolerance was demonstrated in chicken refractory to T. cruzi after the first week of embryonic growth, and chicks hatched from the flagellate-inoculated eggs were unable to produce the specific antibody.&#160;Moreover, the instillation of the human semen ejaculates intraperitoneally or into the vagina of naive mice yielded T. cruzi amastigotes in the epididymis, seminiferous tubule, vas deferens and uterine tube with an absence of inflammatory reactions in the immune privileged organs of reproduction. The breeding of T. cruzi-infected male and female mice with naive mates resulted in acquisition of the infections, which were later transmitted to the progeny. Therefore, a robust education, information and communication program that involves the population and social organizations is deemed necessary to prevent Chagas disease.

The Trypanosoma cruzi agent of Chagas disease produces long-lasting asymptomatic infections that abruptly develop into clinically recognized pathology. The following research protocol describes a short-run family-based epidemiological study to unravel the T. cruzi infection transmitted sexually from parent to progeny.

美国锥虫从男性和女性到天真的命运的性传播

American trypanosomiasis is transmitted to humans by triatomine bugs through the ingestion of contaminated food, by blood transfusions or accidently in ...

Assessment of Child Anthropometry in a Large Epidemiologic Study

A high proportion of children have overweight and obesity in the United States and other countries. Accurate assessment of anthropometry is essential to understand health effects of child growth and adiposity. Gold standard methods of measuring adiposity, such as dual X-ray absorptiometry (DXA), may not be feasible in large field studies. Research staff can, however, complete anthropometric measurements, such as body circumferences and skinfold measurements, using inexpensive portable equipment. In this protocol we detail how to obtain manual anthropometric measurements from children, including standing and sitting height, weight, waist circumference, hip circumference, mid-upper arm circumference, triceps skinfold thickness, and subscapular skinfold thickness, and procedures to assess the quality of these measurements. To demonstrate accuracy of these measurements, among 1,110 school-aged children in the pre-birth cohort Project Viva we calculated Spearman correlation coefficients comparing manual anthropometric measurements with a gold standard measure of body fat, DXA fat mass1. To address reliability, we evaluate intra-rater technical error of measurement at a quality control session conducted on adult female volunteers.

In epidemiologic studies of children, well-trained research staff can accurately and precisely assess weight, height, sitting height, skinfold thicknesses, and body circumferences.

大型流行病学研究儿童人体测量评估

A high proportion of children have overweight and obesity in the United States and other countries. Accurate assessment of anthropometry is essential to ...

医学

Standard Membrane Feeding Assay for the Detection of Plasmodium falciparum Infection in Anopheles Mosquito Vectors

Malaria remains one of the most devastating diseases worldwide and, to date, the African region is still responsible for 94% of all cases worldwide. This parasitic disease requires a protozoan parasite, an Anopheles mosquito vector, and a vertebrate host. The Anopheles genus comprises more than 500 species, of which 60 are known as vectors of the parasite. The Plasmodium parasite genus consists of 250 species, and 48 of these are involved in disease transmission. Furthermore, the Plasmodium falciparum parasite has contributed toward an estimated 99.7% of malaria cases in sub-Saharan Africa in recent years. 
Gametocytes form part of the sexual stage of the parasite and are ingested by the female mosquito upon feeding on an infected human host. Further development of the parasite within the mosquito is enhanced by favorable environmental conditions in the midgut of the mosquito. Here, the fusion of the female and male gametes takes place, and the motile ookinetes originate. The ookinetes enter the midgut epithelium of the mosquito, and mature ookinetes form oocysts, which, in turn, produce motile sporozoites. These sporozoites migrate to the mosquito's salivary glands and are injected as a mosquito takes a blood meal. 
For drug discovery purposes, mosquitoes were artificially infected with gametocyte-infected blood in the standard membrane feeding assay (SMFA). To detect infection within the mosquito and/or to assess the efficacy of antimalarial compounds, the midguts of the female mosquitoes were removed post infection and were stained with mercurochrome. This method was used to enhance the visual detection of oocysts under the microscope for the accurate determination of oocyst prevalence and intensity.

Standard Membrane Feeding Assay for the Detection of Plasmodium falciparum Infection in Anopheles Mosquito Vectors

The standard membrane feeding assay (SMFA) is regarded as the gold standard for the assessment and identification of potential antimalarial compounds. This artificial feeding system is used to infect mosquitoes to further evaluate the effects of such compounds on the intensity and prevalence of the Plasmodium falciparum parasite.

用于检测按蚊媒介中恶性疟原虫感染的标准膜喂养测定

Malaria remains one of the most devastating diseases worldwide and, to date, the African region is still responsible for 94% of all cases worldwide. This ...

A Strategy for the Study of IL-9-Producing Lymphoid Cells in the Nippostrongylus brasiliensis Infection Model

IL-9 is a pleiotropic cytokine associated with various processes, including antitumor immunity, induction of allergic pathologies, and the immune response against helminth infections, where it plays an important role in the expulsion of the parasite. In a murine model of Nippostrongylus brasiliensis infection, IL-9 is produced mainly by CD4+ T lymphocytes and innate lymphoid cells found in the lung, small intestine, and draining lymph nodes. Given the technical difficulties involved in the intracellular staining of IL-9, as well as the complexity of isolating hematopoietic cells from the small intestine upon infection, there is a pressing need for a comprehensive but straightforward protocol to analyze the expression of IL-9 in different lymphoid and non-lymphoid tissues in this model. The protocol described here outlines the kinetics of IL-9 produced by CD4+ T cells and innate lymphoid cells in the lung and small intestine, the main organs targeted by N. brasiliensis, as well as in the mediastinal and mesenteric lymph nodes, throughout the infection. In addition, it details the number of larvae needed for infection, depending on the cell type and organ of interest. This protocol aims to assist in the standardization of assays to save time and resources by offering the opportunity to focus on the specific cells, organs, and disease stages of interest in the N. brasiliensis infection model.

A Strategy for the Study of IL-9-Producing Lymphoid Cells in the Nippostrongylus brasiliensis Infection Model

IL-9-expressing T and ILC2 cells are induced during N. brasiliensis infection, yet their characterization has been largely overlooked in the infected intestine due to their low frequency and differential kinetics. This protocol describes the isolation of these cells from different target organs and confirmation of their identity via flow cytometry at different infection stages.

巴西黑线虫感染模型中产生IL-9的淋巴细胞研究策略

IL-9 is a pleiotropic cytokine associated with various processes, including antitumor immunity, induction of allergic pathologies, and the immune response ...

NAD(P)H Fluorescence Lifetime Imaging for the Metabolic Analysis of the Murine Intestine and Parasites During Nematode Infection

Parasites generally have a negative effect on the health of their host. They represent a huge health burden, as they globally affect the health of the infested human or animal in the long term and, thus, impact agricultural and socio-economic outcomes. However, parasite-driven immune-regulatory effects have been described, with potential therapeutic relevance for autoimmune diseases. While the metabolism in both the host and parasites contributes to their defense and is the basis for nematode survival in the intestine, it has remained largely understudied due to a lack of adequate technologies. We have developed and applied NAD(P)H fluorescence lifetime imaging to explanted murine intestinal tissue during infection with the natural nematode Heligmosomoides polygyrus to study the metabolic processes in both the host and parasites in a spatially resolved manner. The exploitation of the fluorescence lifetime of the co-enzymes nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate (NADPH), hereafter NAD(P)H, which are preserved across species, depends on their binding status and the binding site on the enzymes catalyzing metabolic processes. Focusing on the most abundantly expressed NAD(P)H-dependent enzymes, the metabolic pathways associated with anaerobic glycolysis, oxidative phosphorylation/aerobic glycolysis, and NOX-based oxidative burst, as a major defense mechanism, were distinguished, and the metabolic crosstalk between the host and parasite during infection was characterized.

NADPH Fluorescence Lifetime Imaging for the Metabolic Analysis of the Murine Intestine and Parasites During Nematode Infection

The present protocol describes the NAD(P)H fluorescence lifetime imaging of an explanted murine intestine infected with the natural parasite Heligmosomoides polygyrus, which allows one to investigate metabolic processes both in host and parasite tissues in a spatially resolved manner.

Parasites generally have a negative effect on the health of their host. They represent a huge health burden, as they globally affect the health of the ...

通过粪便漂浮技术识别人畜共患蛔虫

A Simple Fecal Flotation Method for Diagnosing Zoonotic Nematodes Under Field and Laboratory Conditions

Diagnosis of canine parasites with zoonotic potential such as Toxocara canis and Ancylostoma caninum under field conditions is usually difficult due to limited access to a laboratory in rural and suburban areas in Mexico. This study aimed to detect T. canis and Ancylostoma spp. in fecal samples collected from dogs in Mexico from 2017 to 2021 under field conditions. Sample size calculation resulted in a target enrollment of 534 dogs across the country.
Samples were collected directly from the rectum or the ground after defecation. Samples were stored in individual, tightly sealed, plastic bags at 4 &#176;C. A saturated solution of sodium chloride (specific gravity [SpG] 1.20) was prepared both under field and laboratory conditions. Within 3 days of collection, 2-4 g of feces were tested for parasites using a flotation method by suspending each fecal sample in a saline solution. Feces were mixed with the flotation solution and crushed using a metal spoon.
Once a uniform consistency was achieved, the fecal sample was poured into a new plastic cup using a sieve and allowed to sit for 10-15 min. Three drops from the top of the mixture were collected using a sterilized inoculating loop. The slides were placed on the microscope and parasites were identified by trained parasitologists. Fecal samples from 1,055 dogs were screened microscopically. The number of positive samples for Ancylostoma spp.&#160;was 833 (78.95% frequency) and 222 (21.04%) for T. canis. These findings illustrate the importance of identifying zoonotic helminths in dogs living in urban and rural areas in Mexico using a coproparasitoscopic technique in the laboratory and under field conditions.

作者聚焦： 狗粪便的副寄生虫镜检查用于控制和预防人畜共患寄生虫病

This work describes the use of a flotation method to identify Toxocara canis and Ancylostoma spp. detected in fecal samples collected from dogs in Mexico from 2017 to 2021 under field conditions.

一种在野外和实验室条件下诊断人畜共患线虫的简单粪便浮选方法

Diagnosis of canine parasites with zoonotic potential such as Toxocara canis and Ancylostoma caninum under field conditions is usually difficult due to ...