The detection and identification of pathogens from water samples remain challenging due to variations in recovery rates and the cost of procedures. Ultrafiltration offers the possibility to concentrate viral, bacterial, and protozoan organisms in a single process by using size-exclusion-based filtration. In this study, two hollow-fiber ultrafilters with 50,000-molecular-weight cutoffs were evaluated to concentrate microorganisms from 2-and 10-liter water samples. When known quantities (10 5 to 10 6 CFU/liter) of two species of enteric bacteria were introduced and concentrated from 2 liters of sterile water, the addition of 0.1% Tween 80 increased Escherichia coli strain K-12 recoveries from 70 to 84% and Salmonella enterica serovar Enteritidis recoveries from 36 to 72%. An E. coli antibiotic-resistant strain, XL1-Blue, was recovered at a level (87%) similar to that for strain K-12 (96%) from 10 liters of sterile water. When E. coli XL1-Blue was introduced into 10 liters of nonsterile Rio Grande water with higher turbidity levels (23 to 29 nephelometric turbidity units) at two inoculum levels (9 ؋ 10 5 and 2.4 ؋ 10 3 per liter), the recovery efficiencies were 89 and 92%, respectively. The simultaneous addition of E. coli XL1-Blue (9 ؋ 10 5 CFU/liter), Cryptosporidium parvum oocysts (10 oocysts/liter), phage T1 (10 5 PFU/liter), and phage PP7 (10 5 PFU/liter) to 10 liters of Rio Grande surface water resulted in mean recoveries of 96, 54, 59, and 46%, respectively. Using a variety of surface waters from around the United States, we obtained recovery efficiencies for bacteria and viruses that were similar to those observed with the Rio Grande samples, but recovery of Cryptosporidium oocysts was decreased, averaging 32% (the site of collection of these samples had previously been identified as problematic for oocyst recovery). Results indicate that the use of ultrafiltration for simultaneous recovery of bacterial, viral, and protozoan pathogens from variable surface waters is ready for field deployment.

Central Asia is a vast geographic region that includes five former Soviet Union republics: Kazakhstan, Kyrgyzstan, Tajikistan, Turkmenistan, and Uzbekistan. The region has a unique infectious disease burden, and a history that includes Silk Road trade routes and networks that were part of the anti-plague and biowarfare programs in the former Soviet Union. Post-Soviet Union biosurveillance research in this unique area of the world has met with several challenges, including lack of funding and resources to independently conduct hypothesis driven, peer-review quality research. Strides have been made, however, to increase scientific engagement and capability. Kazakhstan and Kyrgyzstan are examples of countries where biosurveillance research has been successfully conducted, particularly with respect to especially dangerous pathogens. In this review, we describe in detail the successes, challenges, and opportunities of conducting biosurveillance in Central Asia as exemplified by our recent research activities on ticks and tick-borne diseases in Kazakhstan and Kyrgyzstan.

Strategic laboratory planning in limited resource areas is essential for addressing global health security issues. Establishing a national reference laboratory, especially one with BSL-3 or -4 biocontainment facilities, requires a heavy investment of resources, a multisectoral approach, and commitments from multiple stakeholders. We make the case for donor organizations and recipient partners to develop a comprehensive laboratory operations roadmap that addresses factors such as mission and roles, engaging national and political support, securing financial support, defining stakeholder involvement, fostering partnerships, and building trust. Successful development occurred with projects in African countries and in Azerbaijan, where strong leadership and a clear management framework have been key to success. A clearly identified and agreed management framework facilitate identifying the responsibility for developing laboratory capabilities and support services, including biosafety and biosecurity, quality assurance, equipment maintenance, supply chain establishment, staff certification and training, retention of human resources, and sustainable operating revenue. These capabilities and support services pose rate-limiting yet necessary challenges. Laboratory capabilities depend on mission and role, as determined by all stakeholders, and demonstrate the need for relevant metrics to monitor the success of the laboratory, including support for internal and external audits. Our analysis concludes that alternative frameworks for success exist for developing and implementing capabilities at regional and national levels in limited resource areas. Thus, achieving a balance for standardizing practices between local procedures and accepted international standards is a prerequisite for integrating new facilities into a country's existing public health infrastructure and into the overall international scientific community.

drain), inability to maintain equipment through service contracts and difficulty in obtaining new state-of-the-art equipment, research kits and technologies. Lacking these resources, research quality will stagnate then regress and, thus, the ability to competitively apply for grant funding will diminish, exacerbating the problem. To maintain an upward cycle, a mechanism to assess and plan for success in obtaining and administering grant funding must be instituted early on, so that mitigation processes can be initiated as challenges are identified.The current lack of an overall strategy and programme structure to assess and evaluate the success of securing and administering research funding will hinder Organisations in under-resourced areas that achieve long-term research sustainability by successfully competing for research funding will not only build their reputation for conducting quality science but also develop their human resources in a manner that reduces the risk of becoming a future security threat. Major challenges to these organisations include identifying and prioritising funding opportunities, securing and administering external grant awards and publishing both the outcomes of research and relevant surveillance data. Lack of a standardised evaluation technique to assess institutional research capabilities poses challenges for identifying and targeting specific, repeatable processes that lead to organisational improvements. Short-and long-term goals, which are challenged by research quality, funding and human resources, need to be established in order to achieve complex missions such as reducing global health security threats. Once baseline capabilities are established, a consistent evaluation technique provides an objective view to complement other steps that enhance capabilities. The capability maturity model, which is often used in business and technology sectors for establishing life cycle and planning sustainment, is a technique that enhances performance by defining three levels of capability (initial, managed and optimised). An organisation can assess its current state of capability ('as is') and develop an actionable strategy for its next progression ('to be'). In addition, application of a CMM aids creation of a strategy for realising a more repeatable and optimised process. Research programmes frequently rely on basic metrics such as the number of peer-reviewed publications and grant funding awards to measure their quality. Our analysis suggests an approach that includes references and tools, especially those that are risk-based, which can be used to establish initial best practices, define metrics, measure outputs and rates of success in a stepwise manner. In addition, we provide a pilot example from a survey of research institutes in under-resourced areas. KEYWORDScapability Maturity Model (cMM); global health security; research ARTICLE HISTORY

The World Health Organization (WHO) estimates that zoonotic diseases transmitted from animals to humans account for 75 percent of new and emerging infectious diseases. Globally, high-consequence pathogens that impact livestock and have the potential for human transmission create research paradoxes and operational challenges for the high-containment laboratories that conduct work with them. These specialized facilities are required for conducting all phases of research on high-consequence pathogens (basic, applied, and translational) with an emphasis on both the generation of fundamental knowledge and product development. To achieve this research mission, a highly-trained workforce is required and flexible operational methods are needed. In addition, working with certain pathogens requires compliance with regulations such as the Centers for Disease Control (CDC) and the U.S. Department of Agriculture (USDA) Select Agent regulations, which adds to the operational burden. The vast experience from the existing studies at Plum Island Animal Disease Center, other U.S. laboratories, and those in Europe and Australia with biosafety level 4 (BSL-4) facilities designed for large animals, clearly demonstrates the valuable contribution this capability brings to the efforts to detect, prepare, prevent and respond to livestock and potential zoonotic threats. To raise awareness of these challenges, which include biosafety and biosecurity issues, we held a workshop at the 2018 American Society for Microbiology (ASM) Biothreats conference to further discuss the topic with invited experts and audience participants. The workshop covered the subjects of research funding and metrics, economic sustainment of drug and vaccine development pipelines, workforce turnover, and the challenges of maintaining operational readiness of high containment laboratories.