13TSB_ENDANI Development of a pen-side test for liver fluke in cattle and sheep

13TSB_ENDANI 牛羊肝吸虫圈侧测试的开发

• 批准号: BB/L011530/1
• 负责人: Kevin Gough
• 金额: $ 22.67万
• 依托单位: University of Nottingham
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FL011530%2F1
• 关键词: 13TSB_ENDANI; Development; pen; side; test

Fasciolosis is a major disease of cattle and sheep and is a key problem within the farming industry. It is caused by the parasite, Fasciola hepatica (a liver fluke), and is acquired by the ingestion of water or vegetation contaminated by the infectious stage of the parasite. The parasite goes on to colonise the liver of cattle and sheep, following migration across the gut wall and peritoneal cavity, where they feed on liver cells and blood. The presence of these parasites causes reduced anaemia, poor liver function, yields, poor fertility and high perinatal loses, and chronic weight loss in cattle. In addition to these effects seen in cattle, acute disease can cause sudden death from haemorrhage and liver damage in sheep. These effects in turn cause economic losses, estimated to be £1312 million worldwide. Currently diagnosis of fasciolosis is based upon epidemiological data and blood samples (for raised liver enzymes) in the acute/sub-acute disease, and the demonstration of eggs in faeces in chronic disease. A number of enzyme-linked immunosorbent assay (ELISA) techniques have also been developed, however, both these and egg counts have to be carried out in central laboratories with the expertise to do so and can take several days to perform, thus there is a clear need for a simple rapid decentralised test. Additionally, current serology based methods of diagnosis fail to distinguish between previous and current infection, while the faecal egg count is both time and labour intensive. Control of infection following diagnosis isthrough the use of flukicides, the efficacy of these drugs is currently under question as drug resistance is now a significant problem for many farmers. Furthermore, the use of classes of these drugs is also now prohibited in dairy animals producing milk for the human food chain. Ultimately this means a more evidence and rational approach to drug usage is needed to prevent further drug resistance and concerns for human food safety. To do would require a test capable of distinguishing not only current from previous infection but also staging the infection in terms of mature versus immature fluke being present. This project will aim to develop a pen-side lateral flow device (LFD) for the diagnosis of fasciolosis in cattle and sheep from faecal samples, with the aim of producing a simple to use, rapid test that will allow farmers and veterinarians to make informed decisions on whether treatment is required.We will achieve this through the combined used of the project partners expertise to complete the following aims; 1) Preparation of F. hepatica antigens (proteins) for immunising mice to generate antibody producing cells; 2) Creation of immortal hybridomas in vitro producing antibody to the F. hepatica antigens; 3) Optimise ELISA immunoassays using antibodies produced in WP3 to F. hepatica antigens capable of distinguishing mature from immature flukes; 4) Develop and produce prototype penside lateral flow devices; 5) Evaluate prototype lateral flow devices on farms with a current fluke problem or farms with no history of fluke infection . The resulting lateral flow device will be used for simple, rapid diagnosisof fasciolosis at the pen-side and will provide evidence of the life-stage of F. hepatica, this is of paramount importance as it influences the choice and efficacy of drug treatments. In diagnosing the parasite and the most efficacious treatment option the novel test will provide a clear economic benefit to the livestock producer. The test will also allow compliance with government policy on the best practice for F. hepatica treatment being on an individual animal basis. In doing so overall drug use will be reduced again reducing costs to the farmer and contributing to the reduction of anthelmintic-resistance.

片形吸虫病是牛羊的主要疾病，是畜牧业中的一个关键问题。它是由寄生虫肝片吸虫（一种肝吸虫）引起的，通过摄入被寄生虫感染阶段污染的水或植物而获得。这种寄生虫继续在牛和羊的肝脏中定居，然后穿过肠壁和腹膜腔，在那里它们以肝细胞和血液为食。这些寄生虫的存在导致牛贫血减少、肝功能差、产量低、生育力差和围产期损失高以及慢性体重减轻。除了在牛身上看到的这些影响外，急性疾病还可能导致绵羊因出血和肝损伤而突然死亡。这些影响反过来又造成经济损失，估计全球范围内为13.12亿英镑。目前，肝片形吸虫病的诊断是基于流行病学数据和血液样本（肝酶升高）在急性/亚急性疾病，并在慢性疾病的粪便中的鸡蛋的证明。还开发了一些酶联免疫吸附测定（ELISA）技术，但是，这些技术和鸡蛋计数都必须在具有专业知识的中心实验室进行，并且可能需要几天时间才能完成，因此显然需要一种简单快速的分散测试。此外，目前基于血清学的诊断方法无法区分先前和当前感染，而粪便虫卵计数既费时又费力。在诊断后控制感染是通过使用杀吸虫剂，这些药物的有效性目前受到质疑，因为抗药性现在是许多农民的一个重要问题。此外，现在也禁止在为人类食物链生产牛奶的奶畜中使用这些药物。最终，这意味着需要一种更有证据和更合理的药物使用方法，以防止进一步的耐药性和对人类食品安全的担忧。要做到这一点，需要一种测试，不仅能够区分当前感染与之前的感染，而且还能够根据存在的成熟与不成熟的侥幸来对感染进行分期。本项目旨在开发一种用于从粪便样本中诊断牛羊肝片吸虫病的围栏侧流装置（LFD），目的是生产一种简单易用的快速检测方法，使农民和兽医能够就是否需要治疗做出明智的决定。我们将通过联合使用项目合作伙伴的专业知识来实现以下目标：1）制备F.肝纤维化抗原（蛋白质）用于免疫小鼠以产生抗体产生细胞; 2）在体外产生针对肝纤维化的抗体的永生杂交瘤的产生; 3）利用WP 3中产生的抗F.能够区分成熟和未成熟吸虫的肝抗原; 4）开发和生产原型Penside侧流装置; 5）在当前有吸虫问题的农场或没有吸虫感染史的农场上评估原型侧流装置。由此产生的侧流装置将用于简单，快速的诊断fasciolosis在笔侧，并将提供证据的生活阶段的F。对于肝细胞癌，这是至关重要的，因为它影响药物治疗的选择和功效。在诊断寄生虫和最有效的治疗方案中，新的测试将为牲畜生产者提供明显的经济效益。该测试还将允许遵守政府关于F.肝病治疗是基于个体动物的。这样做，药物的总体使用将再次减少，降低农民的成本，并有助于减少驱虫剂的耐药性。

项目成果

A rapid IL-17 response to Cryptosporidium parvum in the bovine intestine.

• DOI: 10.1016/j.vetimm.2017.07.009
• 发表时间: 2017-09
• 期刊: Veterinary immunology and immunopathology
• 影响因子: 1.8
• 作者: Drinkall E;Wass MJ;Coffey TJ;Flynn RJ
• 通讯作者: Flynn RJ

Fasciola hepatica infection reduces Mycobacterium bovis burden and mycobacterial uptake and suppresses the pro-inflammatory response.

• DOI: 10.1111/pim.12326
• 发表时间: 2016-07
• 期刊: Parasite immunology
• 影响因子: 2.2
• 作者: Garza-Cuartero L;O'Sullivan J;Blanco A;McNair J;Welsh M;Flynn RJ;Williams D;Diggle P;Cassidy J;Mulcahy G
• 通讯作者: Mulcahy G

The Chronic Stages of Bovine Fasciola hepatica Are Dominated by CD4 T-Cell Exhaustion.

牛fasciola Hepatica的慢性阶段以CD4 T细胞耗尽为主。

• DOI: 10.3389/fimmu.2017.01002
• 发表时间: 2017
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Sachdev D;Gough KC;Flynn RJ
• 通讯作者: Flynn RJ