Anthrax threatens human and animal health, and people's livelihoods in many rural communities in Africa and Asia. In these areas, anthrax surveillance is challenged by a lack of tools for on-site detection. Furthermore, cultural practices and infrastructure may affect sample availability and quality. Practical yet accurate diagnostic solutions are greatly needed to quantify anthrax impacts. We validated microscopic and molecular methods for the detection of Bacillus anthracis in field-collected blood smears and identified alternative samples suitable for anthrax confirmation in the absence of blood smears. We investigated livestock mortalities suspected to be caused by anthrax in northern Tanzania. Field-prepared blood smears (n = 152) were tested by microscopy using four staining techniques as well as polymerase chain reaction (PCR) followed by Bayesian latent class analysis. Median sensitivity (91%, CI 95% [84-96%]) and specificity (99%, CI 95% [96-100%]) of microscopy using azure B were comparable to those of the recommended standard, polychrome methylene blue, PMB (92%, CI 95% [84-97%] and 98%, CI 95% [95-100%], respectively), but azure B is more available and convenient. Other commonly-used stains performed poorly. Blood smears could be obtained for <50% of suspected anthrax cases due to local customs and conditions. However, PCR on DNA extracts from skin, which was almost always available, had high sensitivity and specificity (95%, CI 95% [90-98%] and 95%, CI 95% [87-99%], respectively), even after extended storage at ambient temperature. Azure B microscopy represents an accurate diagnostic test for animal anthrax that can be performed with basic laboratory infrastructure and in the field. When blood smears are unavailable, PCR using skin tissues provides a valuable alternative for confirmation. Our findings lead to a practical diagnostic approach for anthrax in low-resource settings that can support surveillance and control efforts for anthrax-endemic countries globally.

Aims: This study investigated the efficacy of hydrogen peroxide vapour (HPV) at inactivating hazard group 3 bacteria that have been presented dried from their growth medium to present a realistic challenge. Methods and Results: Hydrogen peroxide vapour technology (Bioquell) was used to decontaminate a class III microbiological safety cabinet containing biological indicators (BIs) made by drying standard working suspensions of the following agents: Bacillus anthracis (Ames) spores, Brucella abortus (strain S99), Burkholderia pseudomallei (NCTC 12939), Escherichia coli O157 ST11 (NCTC 12079), Mycobacterium tuberculosis (strain H37Rv) and Yersinia pestis (strain CO92) on stainless steel coupons. Extended cycles were used to expose the agents for 90 min. The HPV cycle completely inactivated B. anthracis spores, B. abortus, B. pseudomallei, E. coli O157 and Y. pestis when BIs were processed using quantitative and qualitative methods. Whilst M. tuberculosis was not completely inactivated, it was reduced by 4 log 10 from a starting concentration of 10 6 colony-forming units.Conclusions: This study demonstrates that HPV is able to inactivate a range of HG3 agents at high concentrations with associated organic matter, but M. tuberculosis showed increased resistance to the process. Significance and Impact of the Study: This publication demonstrates that HPV can inactivate HG3 agents that have an organic load associated with them. It also shows that M. tuberculosis has higher resistance to HPV than other agents. This shows that an appropriate BI to represent the agent of interest should be chosen to demonstrate a decontamination is successful.

As part of the UK response to the 2013-2016 Ebola virus disease (EVD) epidemic in West Africa, Public Health England (PHE) were tasked with establishing three field Ebola virus (EBOV) diagnostic laboratories in Sierra Leone by the UK Department for International Development (DFID). These provided diagnostic support to the Ebola Treatment Centre (ETC) facilities located in Kerry Town, Makeni and Port Loko. The Novel and Dangerous Pathogens (NADP) Training group at PHE, Porton Down, designed and implemented a pre-deployment Ebola diagnostic laboratory training programme for UK volunteer scientists being deployed to the PHE EVD laboratories. Here, we describe the training, workflow and capabilities of these field laboratories for use in response to disease epidemics and in epidemiological surveillance. We discuss the training outcomes, the laboratory outputs, lessons learned and the legacy value of the support provided. We hope this information will assist in the recruitment and training of staff for future responses and in the design and implementation of rapid deployment diagnostic field laboratories for future outbreaks of high consequence pathogens.This article is part of the themed issue 'The 2013-2016 West African Ebola epidemic: data, decision-making and disease control'.

23Background: Anthrax threatens human and animal health, and people's livelihoods in 24 many rural communities in Africa and Asia. In these areas, anthrax surveillance is 25 challenged by a lack of tools for on-site detection. Furthermore, cultural practices and 26 infrastructure may affect sample availability and quality. Practical yet accurate diagnostic 27 solutions are greatly needed to quantify anthrax impacts. We validated microscopic and 28 molecular methods for the detection of Bacillus anthracis in field-collected blood smears 29 and identified alternative samples suitable for anthrax confirmation in the absence of blood 30 smears. 31 Methodology/Principal Findings:We investigated livestock mortalities suspected to be 32 caused by anthrax in northern Tanzania. Field-prepared blood smears (n = 152) were tested 33 by microscopy using four staining techniques as well as polymerase chain reaction (PCR) 34 followed by Bayesian latent class analysis. Median sensitivity (91%, CI 95% [84-96%]) and 35 specificity (99%, CI 95% [96-100%]) of microscopy using azure B were comparable to 36 those of the recommended standard, polychrome methylene blue, PMB (92%, CI 95% [84-37 97%] and 98%, CI 95% [95-100%], respectively), but azure B is more available and 38 convenient. Other commonly-used stains performed poorly. Blood smears could be 39 obtained for <50% of suspected anthrax cases due to local customs and conditions. 40 However, PCR on DNA extracts from dried skin, which was almost always available, had 41 high sensitivity and specificity (95%, CI 95% [90-98%] and 95%, CI 95% [87-99%], 42 respectively), even after extended storage at ambient temperature. 43 Conclusions/Significance: Azure B microscopy represents an accurate diagnostic test for 44animal anthrax that can be performed with basic laboratory infrastructure and in the field. 45When blood smears are unavailable, PCR using skin tissues provides a valuable alternative 46 for confirmation. Our findings lead to a practical diagnostic approach for anthrax in low-47 resource settings that can support surveillance and control efforts for anthrax-endemic 48 countries globally. 49 Author summary50 Anthrax, an ancient disease largely controlled in the developed world, is still widespread in 51 remote and rural communities of low-and middle-income countries where it affects human 52 and animal health, and livelihoods. To control anthrax effectively, detection and accurate 53 confirmation are important, but solutions need to be feasible for the most-affected areas 54 3 where resources and infrastructure are typically limited. To achieve this, we assessed a 55 newly proposed stain, azure B, for microscopic confirmation on animal blood smears, as 56 this method can be implemented in low-resource laboratories and in the field. Microscopy 57 using azure B was highly accurate compared to other recommended stains and has the 58 added advantage of being more readily available and convenient. However, blood smear 59 samples were unavailable for more than half of suspected case...