Network Experiences from a Cross-Sector Biosafety Level-3 Laboratory Collaboration: A Swedish Forum for Biopreparedness Diagnostics.

The Swedish Forum for Biopreparedness Diagnostics (FBD) is a network that fosters collaboration among the 4 agencies with responsibility for the laboratory diagnostics of high-consequence pathogens, covering animal health and feed safety, food safety, public health and biodefense, and security. The aim of the network is to strengthen capabilities and capacities for diagnostics at the national biosafety level-3 (BSL-3) laboratories to improve Sweden's biopreparedness, in line with recommendations from the EU and WHO. Since forming in 2007, the FBD network has contributed to the harmonization of diagnostic methods, equipment, quality assurance protocols, and biosafety practices among the national BSL-3 laboratories.

One health security: an important component of the global health security agenda.

The objectives of the Global Health Security Agenda (GHSA) will require not only a "One Health" approach to counter natural disease threats against humans, animals, and the environment, but also a security focus to counter deliberate threats to human, animal, and agricultural health and to nations' economies. We have termed this merged approach "One Health Security." It will require the integration of professionals with expertise in security, law enforcement, and intelligence to join the veterinary, agricultural, environmental, and human health experts essential to One Health and the GHSA.

A decade with nucleic acid-based microbiological methods in safety control of foods.

In the last decade, nucleic acid-based methods gradually started to replace or complement the culture-based methods and immunochemical assays in routine laboratories involved in food control. In particular, real-time polymerase chain reaction (PCR) was technically developed to the stage of good speed, sensitivity and reproducibility, at minimized risk of carry-over contamination. Basic advantages provided by nucleic acid-based methods are higher speed and added information, such as subspecies identification, information on the presence of genes important for virulence or antibiotic resistance.

Pre-PCR processing in bioterrorism preparedness: improved diagnostic capabilities for laboratory response networks.

Diagnostic DNA analysis using polymerase chain reaction (PCR) has become a valuable tool for rapid detection of biothreat agents. However, analysis is often challenging because of the limited size, quality, and purity of the biological target. Pre-PCR processing is an integrated concept in which the issues of analytical limit of detection and simplicity for automation are addressed in all steps leading up to PCR amplification--that is, sampling, sample treatment, and the chemical composition of PCR.

The need for high-quality whole-genome sequence databases in microbial forensics.

Microbial forensics is an important part of a strengthened capability to respond to biocrime and bioterrorism incidents to aid in the complex task of distinguishing between natural outbreaks and deliberate acts. The goal of a microbial forensic investigation is to identify and criminally prosecute those responsible for a biological attack, and it involves a detailed analysis of the weapon--that is, the pathogen. The recent development of next-generation sequencing (NGS) technologies has greatly increased the resolution that can be achieved in microbial forensic analyses.

Actionable knowledge and strategic decision making for bio- and agroterrorism threats: building a collaborative early warning culture.

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.

Harmonization of European laboratory response networks by implementing CWA 15793: use of a gap analysis and an "insider" exercise as tools.

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.

Separated by a common language: awareness of term usage differences between languages and disciplines in biopreparedness.

Preparedness for bioterrorism is based on communication between people in organizations who are educated and trained in several disciplines, including law enforcement, health, and science. Various backgrounds, cultures, and vocabularies generate difficulties in understanding and interpretating terms and concepts, which may impair communication. This is especially true in emergency situations, in which the need for clarity and consistency is vital.

Social media and its dual use in biopreparedness: communication and visualization tools in an animal bioterrorism incident.

This article focuses on social media and interactive challenges for emergency organizations during a bioterrorism or agroterrorism incident, and it outlines the dual-use dilemma of social media. Attackers or terrorists can use social media as their modus operandi, and defenders, including emergency organizations in law enforcement and public and animal health, can use it for peaceful purposes. To get a better understanding of the uses of social media in these situations, a workshop was arranged in Stockholm, Sweden, to raise awareness about social media and animal bioterrorism threats.

Agroterrorism targeting livestock: a review with a focus on early detection systems.

Agroterrorism targeting livestock can be described as the intentional introduction of an animal disease agent against livestock with the purpose of causing economic damage, disrupting socioeconomic stability of a country, and creating panic and distress. This type of terrorism can be alluring to terrorists because animal disease agents are easily available. This review addresses the vulnerabilities of the livestock industry to agroterrorism.

Development and optimization of a biopreparedness protocol for extracting and detecting avian influenza virus in broiler chicken meat.

Detection of avian influenza virus (AIV) in poultry meat is hampered by the lack of an efficient analytical method able to extract and concentrate viral RNA prior to PCR. In this study we developed a method for extracting and detecting AIV from poultry meat by a previously standardized 1-step real-time reverse transcriptase PCR (RRT-PCR) assay. In addition, a new process control, represented by feline calicivirus (FCV), was included in the original protocol, to evaluate all analytical steps from sample preparation to the detection phase.

Multiplex real-time PCR for detecting and typing Clostridium botulinum group III organisms and their mosaic variants.

Botulism is a neuroparalytic disease that can occur in all warm-blooded animals, birds, and fishes. The disease in animals is mainly caused by toxins produced by Clostridium botulinum strains belonging to group III, although outbreaks due to toxins produced by group I and II organisms have been recognized. Group III strains are capable of producing botulinum toxins of type C, D, and C/D and D/C mosaic variants.

Historical perspective on agroterrorism: lessons learned from 1945 to 2012.

This article presents a historical perspective on agroterrorism cases from 1945 until 2012. The threat groups and perpetrators associated with bio- and agroterrorism are clustered into several groups: apocalyptic sects, lone wolves, political groups, and religious groups. We used open-source information, and 4 biological agroterrorism cases are described: (1) in 1952, Mau Mau poisoned cattle in Kenya by using a plant toxin from the African milk bush plant; (2) in 1985, the USDA claimed that Mexican contract workers were involved in deliberately spreading screwworm (Cochliomyia hominivorax) among livestock; (3) in 2000, Palestinian media reported that Israeli settlers released sewer water into Palestinian agricultural fields; and (4) in 2011, a person was sentenced to prison after threatening US and UK livestock with the deliberate spread of foot-and-mouth disease virus.

Metagenomic detection methods in biopreparedness outbreak scenarios.

In the field of diagnostic microbiology, rapid molecular methods are critically important for detecting pathogens. With rapid and accurate detection, preventive measures can be put in place early, thereby preventing loss of life and further spread of a disease. From a preparedness perspective, early detection and response are important in order to minimize the consequences.

Towards an international standard for detection and typing botulinum neurotoxin-producing Clostridia types A, B, E and F in food, feed and environmental samples: a European ring trial study to evaluate a real-time PCR assay.

A real-time PCR method for detection and typing of BoNT-producing Clostridia types A, B, E, and F was developed on the framework of the European Research Project "Biotracer". A primary evaluation was carried out using 104 strains and 17 clinical and food samples linked to botulism cases. Results showed 100% relative accuracy, 100% relative sensitivity, 100% relative specificity, and 100% selectivity (inclusivity on 73 strains and exclusivity on 31 strains) of the real-time PCR against the reference cultural method combined with the standard mouse bioassay.

Accidental and deliberate microbiological contamination in the feed and food chains--how biotraceability may improve the response to bioterrorism.

A next frontier of the global food safety agenda has to consider a broad spectrum of bio-risks, such as accidental and intentional contaminations in the food and feed chain. In this article, the background for the research needs related to biotraceability and response to bioterrorism incidents are outlined. Given the current scale of international trade any response need to be considered in an international context.

A multiplex real-time PCR for identifying and differentiating B. anthracis virulent types.

Bacillus anthracis is closely related to the endospore forming bacteria Bacillus cereus and Bacillus thuringiensis. For accurate detection of the life threatening pathogen B. anthracis, it is essential to distinguish between these three species.

Bioinformatic tools for using whole genome sequencing as a rapid high resolution diagnostic typing tool when tracing bioterror organisms in the food and feed chain.

The rapid technological development in the field of parallel sequencing offers new opportunities when tracing and tracking microorganisms in the food and feed chain. If a bioterror organism is deliberately spread it is of crucial importance to get as much information as possible regarding the strain as fast as possible to aid the decision process and select suitable controls, tracing and tracking tools. A lot of efforts have been made to sequence multiple strains of potential bioterror organisms so there is a relatively large set of reference genomes available.

Evaluation of Bacillus strains as model systems for the work on Bacillus anthracis spores.

Available strain collections of Bacillus anthracis and Bacillus cereus were screened for B. cereus strains sharing major genotypic characteristics with B. anthracis.

Real-time PCR for Clostridium botulinum type C neurotoxin (BoNTC) gene, also covering a chimeric C/D sequence--application on outbreaks of botulism in poultry.

In recent years, botulism type C has become a serious problem in poultry flocks in Sweden. A real-time PCR assay for Clostridium botulinum (C. botulinum) type C neurotoxin (BoNTC) gene was developed as an alternative to the mouse bioassay for detection and identification of C.

An innovative molecular detection tool for tracking and tracing Clostridium botulinum types A, B, E, F and other botulinum neurotoxin producing Clostridia based on the GeneDisc cycler.

Rapid and specific detection of botulinum neurotoxin (BoNT) producing Clostridia is a priority for public health authorities, in case of both natural and intentional botulism outbreaks. This study reports on the evaluation of a detection system based on the GeneDisc Cycler designed for simultaneously testing the bont/A, bont/B, bont/E and bont/F genes encoding for the botulinum neurotoxins types A, B, E and F. BoNT-producing Clostridia (n = 102) and non-BoNT-producing bacteria (n = 52) isolated from clinical, food and environmental samples were tested using this macro-array and results were compared to the reference lethality test on mice.

Anthrax outbreak in a Swedish beef cattle herd--1st case in 27 years: Case report.

After 27 years with no detected cases, an outbreak of anthrax occurred in a beef cattle herd in the south of Sweden. The outbreak was unusual as it occurred in winter, in animals not exposed to meat-and-bone meal, in a non-endemic country. The affected herd consisted of 90 animals, including calves and young stock.

Strategies for overcoming PCR inhibition.

INTRODUCTIONThe use of conventional and real-time PCR is to some extent restricted by the presence of PCR inhibitors. This is particularly so when the techniques are applied directly to complex biological samples such as clinical, environmental, or food samples for the detection of microorganisms. PCR inhibitors can originate from the sample itself, or as a result of the method used to collect or otherwise prepare the sample.