Detecting and understanding mycobacterial infections in domestic cats and their role in the risk of zoontoci, intra- and inter-species spread

检测和了解家猫的分枝杆菌感染及其在人畜共患病、物种内和物种间传播风险中的作用

• 批准号: 1658195
• 金额: --
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1658195
• 关键词: Detecting; understanding; mycobacterial; infections; domestic

Mycobacterial infections are a global concern in humans and animals. In the UK, tuberculosis (TB) in animals is not confined to cattle and badgers; it is a serious concern in deer, camelids and cats. We have shown that cats are significantly at risk of TB as ~1% of feline biopsies have changes consistent with mycobacteriosis; 15% due to M. bovis, 19% M. microti (the vole/rodent bacillus). Some M. bovis infections progress extremely rapidly - a major concern given the newly recognised cat to human zoonotic risk, and notifiable status (M. bovis spreads between many different mammalian species). We hypothesise that the outcome of a cats exposure to Mycobacteria is determined by the nature of the pathogen and the host's immune response. However, a major bottleneck is failure to culture in ~50% of cases (despite having ZN+ bacteria). Rapid diagnosis is key to determining zoonotic risk and management options.Currently, identifying the Mycobacteria species depends on a) detecting ZN+ bacteria b) IFN-gamma ELISA c) culture. These tests can detect Mycobacteria and the ELISA differentiates (with some specifity) M. bovis, M. microti and M. avium. Culture is a gold standard method to confirm live bacteria but is lengthy and frequently negative (despite tissue being ZN+). To rapidly identify infecting bacteria will impact significantly on knowing the zoonotic, inter- and intra-species risk, management options, affect survival (likelihood of clearance or latency) and inform ongoing monitoring. It could also monitor in-contact animals. We need to invesitgate the host respone to these infections if we are to understand Mycobacterial diseases more fully, and so elucidate the role of the host's response in determining which cases are likely to spread disease or, where appropriate, respond to treatment.The project has 5 objectives:1) Collect samples from Mycobacteria-infected cats and controls. Confirm Mycobacterial presence using current tests (culture, ZN, IFN-gamma ELISA) (RDSVS, Roslin, Biobest).2) Isolate Mycobacterial DNA using established protocols from fresh frozen tissue or FFPE blocks. Determine the Mycobacterial sequences using the latest second and third generation sequencing methods (Illumina HiSeq 2500/MiSeq; Oxford Nanopore MinION) and compare them to published genome sequences (Roslin/Edinburgh Genomics). 3) Perform comparative analysis of the genome sequences of the isolated strains to identify sequence similarities and differences. Identify unique sequences for the development of strain-specific PCRs and develop multiplex PCR assays for the rapid diagnosis of feline TB and other Mycobacterial infections (Biobest). 4) Monitor cases long-term to correlate specific sequences with disease outcomes, and any transmission (RDSVS).5) The nature of host-pathogen interaction between macrophages and Mycobacteria defines infection outcome in a number of species, but has not been investigated in cats. Understanding the basis for immunity or disease progression will inform veterinary medicine, and help to suggest appropriate management regimens for infected cats. We will identify cats with M. bovis TB and compare macrophage distribution, function and phenotype between infected and control tissues (Roslin).This is a new collaboration bringing together expertise in feline medicine and tuberculosis, with macrophage biology, and pathogen genome sequencing incorporating both Roslin/RDSVS and Biobest Laboratories.

分枝杆菌感染是人类和动物关注的全球性问题。在英国，动物结核病（TB）并不局限于牛和獾；这对鹿、骆驼和猫来说是一个严重的问题。我们已经表明，猫患结核病的风险很大，因为约1%的猫活组织检查有与分枝杆菌病一致的变化；15%为牛支原体，19%为微支原体（田鼠/啮齿动物杆菌）。一些牛支原体感染进展极快——考虑到新发现的猫对人的人畜共患风险和应通报状态（牛支原体在许多不同哺乳动物物种之间传播），这是一个主要问题。我们假设猫暴露于分枝杆菌的结果是由病原体的性质和宿主的免疫反应决定的。然而，一个主要的瓶颈是在约50%的病例中培养失败（尽管有ZN+细菌）。快速诊断是确定人畜共患病风险和管理方案的关键。目前，分枝杆菌种类的鉴定依赖于a)检测ZN+细菌b) ifn - γ ELISA c)培养。这些试验可检测分枝杆菌，ELISA可区分（有一定特异性）牛支原体、微支原体和鸟支原体。培养是确认活菌的金标准方法，但耗时长且经常呈阴性（尽管组织呈ZN+）。快速识别感染细菌将对了解人畜共患病、物种间和物种内风险、管理选择、影响生存（清除或延迟的可能性）以及为持续监测提供信息产生重大影响。它还可以监测接触动物。如果我们要更全面地了解分枝杆菌疾病，我们需要调查宿主对这些感染的反应，从而阐明宿主反应在确定哪些病例可能传播疾病或在适当情况下对治疗有反应方面的作用。该项目有5个目标：1)收集感染分枝杆菌的猫和对照组的样本。使用当前测试（培养、ZN、ifn - γ ELISA） （RDSVS、Roslin、Biobest）确认分枝杆菌的存在。2)使用既定方案从新鲜冷冻组织或FFPE块中分离分枝杆菌DNA。使用最新的第二代和第三代测序方法（Illumina HiSeq 2500/MiSeq; Oxford Nanopore MinION）测定分枝杆菌序列，并将其与已发表的基因组序列（Roslin/Edinburgh Genomics）进行比较。3)对分离菌株的基因组序列进行比较分析，找出序列的异同。鉴定独特的序列，用于开发菌株特异性PCR，并开发多重PCR检测方法，用于快速诊断猫结核病和其他分枝杆菌感染（Biobest）。4)长期监测病例，将特定序列与疾病结局和任何传播（RDSVS）联系起来。5)在许多物种中，巨噬细胞和分枝杆菌之间的宿主-病原体相互作用的性质决定了感染的结果，但尚未在猫中进行研究。了解免疫或疾病进展的基础将为兽医提供信息，并有助于为受感染的猫提出适当的管理方案。我们将鉴定患有牛分枝杆菌结核的猫，并比较感染组织和对照组织之间巨噬细胞的分布、功能和表型（Roslin）。这是一项新的合作，汇集了猫科医学和结核病方面的专业知识，巨噬细胞生物学，以及Roslin/RDSVS和Biobest实验室的病原体基因组测序。

项目成果

An outbreak of tuberculosis due to Mycobacterium bovis infection in a pack of English Foxhounds (2016-2017).

• DOI: 10.1111/tbed.12969
• 发表时间: 2018-12
• 期刊: Transboundary and emerging diseases
• 影响因子: 4.3
• 作者: O'Halloran C;Hope JC;Dobromylskyj M;Burr P;McDonald K;Rhodes S;Roberts T;Dampney R;De la Rua-Domenech R;Robinson N;Gunn-Moore DA
• 通讯作者: Gunn-Moore DA

Tuberculosis joint infections in four domestic cats.

• DOI: 10.1177/2055116917719401
• 发表时间: 2017-07
• 期刊: JFMS open reports
• 作者: Lalor SM;Clarke S;Pink J;Parry A;Scurrell E;Fitzpatrick N;Watson F;O'Halloran C;Gunn-Moore D
• 通讯作者: Gunn-Moore D

Unusual presentations of feline cowpox.

猫科动物牛痘的异常表现。

• DOI: 10.1136/vr.i5767
• 发表时间: 2016
• 期刊: The Veterinary record
• 作者: O'Halloran C

Novel characteristics identified in two cases of feline cowpox virus infection.

• DOI: 10.1177/2055116917717191
• 发表时间: 2017-07
• 期刊: JFMS open reports
• 作者: Breheny CR;Fox V;Tamborini A;O'Halloran C;Robertson E;Cazzini P;Birn-Jeffery D;Henkin J;Schwartz T;Scase T;Powell R;Gunn-Moore D
• 通讯作者: Gunn-Moore D

Bovine tuberculosis in working foxhounds: lessons learned from a complex public health investigation.

• DOI: 10.1017/s0950268818002753
• 发表时间: 2018-10-09
• 期刊: Epidemiology and infection
• 影响因子: 4.2
• 作者: Phipps E;McPhedran K;Edwards D;Russell K;O'Connor CM;Gunn-Moore DA;O'Halloran C;Roberts T;Morris J
• 通讯作者: Morris J