Current trends in biosecurity and cybersecurity include (1) the wide availability of technology and specialized knowledge that previously were available only to governments; (2) the global economic recession, which may increase the spread of radical non-state actors; and (3) recent US and EU commission reports that reflect concerns about non-state actors in asymmetric threats. The intersectoral and international nature of bioterrorism and agroterrorism threats requires collaboration across several sectors including intelligence, police, forensics, customs, and other law enforcement organizations who must work together with public and animal health organizations as well as environmental and social science organizations. This requires coordinated decision making among these organizations, based on actionable knowledge and information sharing. The risk of not sharing information among organizations compared to the benefit of sharing information can be considered in an "information sharing risk-benefit analysis" to prevent a terrorism incident from occurring and to build a rapid response capability. In the EU project AniBioThreat, early warning is the main topic in work package 3 (WP 3). A strategy has been generated based on an iterative approach to bring law enforcement agencies and human and animal health institutes together. Workshops and exercises have taken place during the first half of the project, and spin-off activities include new preparedness plans for institutes and the formation of a legal adviser network for decision making. In addition, a seminar on actionable knowledge was held in Stockholm, Sweden, in 2012, which identified the need to bring various agency cultures together to work on developing a resilient capability to identify early signs of bio- and agroterrorism threats. The seminar concluded that there are a number of challenges in building a collaborative culture, including developing an education program that supports collaboration and shared situational awareness.
The aim of this study was to develop a rapid PCR-based detection method to confirm presumptive isolates of Salmonella spp. as well as to directly detect Salmonella in feed samples. The specificity of the assay was confirmed by testing I 04 Salmonella strains and 43 strains of other Enterobacteriaceae species. A simple sample preparation method was developed based on dilution of suspended cells picked from agar plates after selective enrichment. The method was evaluated on 109 food and feed samples and there was 100% agreement between the traditional method based on biochemical tests and the PCR method. The inhibitory effect on PCR for 15 different feed samples was overcome by using Tth DNA polymerase in combination with addition of glycerol, which enabled direct detection of Salmonella cells in feed.
Laboratory response networks (LRNs) have been established for security reasons in several countries including the Netherlands, France, and Sweden. LRNs function in these countries as a preparedness measure for a coordinated diagnostic response capability in case of a bioterrorism incident or other biocrimes. Generally, these LRNs are organized on a national level. The EU project AniBioThreat has identified the need for an integrated European LRN to strengthen preparedness against animal bioterrorism. One task of the AniBioThreat project is to suggest a plan to implement laboratory biorisk management CWA 15793:2011 (CWA 15793), a management system built on the principle of continual improvement through the Plan-Do-Check-Act (PDCA) cycle. The implementation of CWA 15793 can facilitate trust and credibility in a future European LRN and is an assurance that the work done at the laboratories is performed in a structured way with continuous improvements. As a first step, a gap analysis was performed to establish the current compliance status of biosafety and laboratory biosecurity management with CWA 15793 in 5 AniBioThreat partner institutes in France (ANSES), the Netherlands (CVI and RIVM), and Sweden (SMI and SVA). All 5 partners are national and/or international laboratory reference institutes in the field of public or animal health and possess high-containment laboratories and animal facilities. The gap analysis showed that the participating institutes already have robust biorisk management programs in place, but several gaps were identified that need to be addressed. Despite differences between the participating institutes in their compliance status, these variations are not significant. Biorisk management exercises also have been identified as a useful tool to control compliance status and thereby implementation of CWA 15793. An exercise concerning an insider threat and loss of a biological agent was performed at SVA in the AniBioThreat project to evaluate implementation of the contingency plans and as an activity in the implementation process of CWA 15793. The outcome of the exercise was perceived as very useful, and improvements to enhance biorisk preparedness were identified. Gap analyses and exercises are important, useful activities to facilitate implementation of CWA 15793. The PDCA cycle will enforce a structured way to work, with continual improvements concerning biorisk management activities. Based on the activities in the AniBioThreat project, the following requirements are suggested to promote implementation: support from the top management of the organizations, knowledge about CWA 15793, a compliance audit checklist and gap analysis, training and exercises, networking in LRNs and other networks, and interinstitutional audits. Implementation of CWA 15793 at each institute would strengthen the European animal bioterrorism response capabilities by establishing a well-prepared LRN.
A review describes the sample prepn. for biochem. methods, enrichment methods, immunol. methods, and phys. methods. The pretreatment of a complex biol. sample is crucial, and for successful PCR the following requirements have to be fulfilled: complete lack or low concn. of PCR-inhibitory components in the sample and sufficient concn. of target DNA. The aim of the pre-PCR treatment is to convert a complex biol. sample contg. the target microorganisms into PCR-amplifiable samples.
ABSTRACT A PCR procedure has been developed for routine analysis of viable Salmonella spp. in feed samples. The objective was to develop a simple PCR-compatible enrichment procedure to enable DNA amplification without any sample pretreatment such as DNA extraction or cell lysis. PCR inhibition by 14 different feed samples and natural background flora was circumvented by the use of the DNA polymerase Tth . This DNA polymerase was found to exhibit a high level of resistance to PCR inhibitors present in these feed samples compared to DyNAzyme II, FastStart Taq , Platinum Taq , Pwo , r Tth , Taq , and Tfl . The specificity of the Tth assay was confirmed by testing 101 Salmonella and 43 non- Salmonella strains isolated from feed and food samples. A sample preparation method based on culture enrichment in buffered peptone water and DNA amplification with Tth DNA polymerase was developed. The probability of detecting small numbers of salmonellae in feed, in the presence of natural background flora, was accurately determined and found to follow a logistic regression model. From this model, the probability of detecting 1 CFU per 25 g of feed in artificially contaminated soy samples was calculated and found to be 0.81. The PCR protocol was evaluated on 155 naturally contaminated feed samples and compared to an established culture-based method, NMKL-71. Eight percent of the samples were positive by PCR, compared with 3% with the conventional method. The reasons for the differences in sensitivity are discussed. Use of this method in the routine analysis of animal feed samples would improve safety in the food chain.
