Dissection of the molecular circuitry controlling virulence in Francisella tularensis.

Francisella tularensis, the etiological agent of tularemia, is one of the most infectious bacteria known. Because of its extreme pathogenicity, F. tularensis is classified as a category A bioweapon by the US government. F. tularensis virulence stems from genes encoded on the Francisella pathogenicity island (FPI). An unusual set of Francisella regulators-the heteromeric macrophage growth locus protein A (MglA)-stringent starvation protein A (SspA) complex and the DNA-binding protein pathogenicity island gene regulator (PigR)-activates FPI transcription and thus is essential for virulence. Intriguingly, the second messenger, guanosine-tetraphosphate (ppGpp), which is produced during infection, is also involved in coordinating Francisella virulence; however, its role has been unclear. Here we identify MglA-SspA as a novel ppGpp-binding complex and describe structures of apo- and ppGpp-bound MglA-SspA. We demonstrate that MglA-SspA, which binds RNA polymerase (RNAP), also interacts with the C-terminal domain of PigR, thus anchoring the (MglA-SspA)-RNAP complex to the FPI promoter. Furthermore, we show that MglA-SspA must be bound to ppGpp to mediate high-affinity interactions with PigR. Thus, these studies unveil a novel pathway different from those described previously for regulation of transcription by ppGpp. The data also indicate that F. tularensis pathogenesis is controlled by a highly interconnected molecular circuitry in which the virulence machinery directly senses infection via a small molecule stress signal.

Engaging Communities in Emerging Infectious Disease Mitigation to Improve Public Health and Safety.

The COVID-19 pandemic highlighted the need for potent community-based tools to improve preparedness. We developed a community health-safety climate (HSC) measure to assess readiness to adopt health behaviors during a pandemic. We conducted a mixed-methods study incorporating qualitative methods (e.g., focus groups) to generate items for the measure and quantitative data from a February 2021 national survey to test reliability, multilevel construct, and predictive and nomologic validities. The 20-item HSC measure is unidimensional (Cronbach α = 0.87). All communities had strong health-safety climates but with significant differences between communities (F = 10.65; p<0.001), and HSC levels predicted readiness to adopt health-safety behaviors. HSC strength moderated relationships between HSC level and behavioral indicators; higher climate homogeneity demonstrated stronger correlations. The HSC measure can predict community readiness to adopt health-safety behaviors in communities to inform interventions before diseases spread, providing a valuable tool for public health authorities and policymakers during a pandemic.

Integrating Veterinary Diagnostic Laboratories for Emergency Use Testing during Pandemics1.

The SARS-CoV-2 pandemic showed limitations in human outbreak testing. Veterinary diagnostic laboratories (VDLs) possess capabilities to bolster emergency test capacity. Surveys from 26 participating VDLs found human SARS-CoV-2 testing was mutually beneficial, including One Health benefits. VDLs indicated testing >3.8 million human samples during the pandemic, which included some challenges.

Comparison of nine extraction methods for bacterial identification using the ONT MinION sequencer.

The Anthrax mailings bioterrorism attack in 2001 revealed the need for universal and rapid microbial forensic analyses on unknown biological evidence. However, the gold standard for bacterial identification includes culturing isolates, which is laborious. Molecular approaches for bacterial identification revolve around 16S ribosomal gene sequencing using Sanger or next generation sequencing (NGS) platforms, but these techniques are laboratory-based and can also be time-consuming. The Oxford Nanopore Technologies (ONT) MinION sequencer can generate long read lengths that span the entire bacterial 16S rRNA gene and accurately identify the species level. This platform can be used in the field, allowing on-site evidence analysis. However, it requires higher quantities of pure DNA compared to other sequencing platforms; thus, the extraction method for bacterial DNA is critical for downstream analysis, which to date are tailored toward a priori knowledge of the species' taxonomic grouping. During an attack, the investigative team may not know what species they are handling; therefore, identifying an extraction method that can handle all bacterial groups and generate clean DNA for the MinION is useful for microbial forensic analysis. The purpose of this study was to identify a "universal" extraction method that can be coupled with ONT MinION sequencing for use in forensic situations for rapid identification. It also evaluated the cloud-based data analysis software provided by ONT, EPI2ME. No "universal" extraction method was identified as optimal for downstream MinION sequencing. However, the DNeasy PowerSoil Kit and Noda et al. Chelex-100 method gave comparable sequencing results and could be used as rapid extraction techniques. This study showed that the ONT 16S Barcoding Kit 1-24 coupled with the 16S FASTQ workflow might not be the best for use in forensic situations where species-level identification needs to be obtained, as most alignments were approximately 89% accurate. In all seven test organisms and nine extraction methods, accurate species identification was only obtained in 63% of the cases.

Terbium Phenanthroline Complex as a Luminescent Probe for the Detection of Anthrax Biomarker: Dipicolinic acid.

Dipicolinic acid (DPA) is a prominent biomarker for Anthrax disease. Bacillus anthracis bacterial endospores is composed of DPA as the significant component, which on over inhalation can cause severe health issues. Such contagious and life-threatening pathogens can be employed as bioweapons or biothreat agents for spreading bioterrorism which is a major risk for national security and public health concerns. Hence, effective detection or a surveillance system is essential for preventing the growth of bioterrorism events. Herein, we have developed a Terbium - 1,10 Phenanthroline (Tb-Phen) based lanthanide luminescence complex with bright green fluorescence. On addition of DPA, the green fluorescence is turn-off at a linear range from 0.6 to 4.762 mM. In this effect, 5D4- 7F5 transition caused by 1,10-phenanthroline to Tb3+ at 544 nm is restricted due to energy transfer quenching and Inner Filter Effect (IFE). The developed probe shows good sensitivity towards the detection of DPA with other coexisting biomolecules and ions with a low Limit of Detection (LOD) of 5.029 µM. The practical feasibility was evaluated in paper strip assay and extended in real samples such as human serum and tap water with satisfactory recovery percentage. Thereby, probe finds promising application in sensing of anthrax spore biomarker (DPA) and biothreat agents.

Poxviruses as Agents of Biological Warfare: The Importance of Ensuring Ethical Standards for Research with Viruses.

Historically, biological agents have been used to target various populations. One of the earliest examples could be the catastrophic effect of smallpox in Australia in the eighteenth century (as alleged by some historians). Modern biological techniques can be used to both create or provide protection against various agents of biological warfare. Any microorganism (viruses, bacteria, and fungi) or its toxins can be used as biological agents. Minnesota Department of Health has listed Smallpox (variola major) as a category A bioterrorism agent, even though it has been eradicated in 1980 through an extensive vaccination campaign. Category A agents are considered the highest risk to public health. Laboratory-associated outbreaks of poxviruses could cause unprecedented occupational hazards. Only two WHO-approved BSL-4 facilities in the United States and Russia are allowed to perform research on the variola virus. So, poxviruses present themselves as a classical case of a dual-use dilemma, since research with them can be used for both beneficial and harmful purposes. Although the importance of ethics in scientific research requires no further elaboration, ethical norms assume greater significance during experimentation with poxviruses. In this chapter, we will update the readers on the sensitive nature of conducting research with poxviruses, and how these viruses can be a source of potential biological weapons. Finally, specified ethical guidelines are explored to ensure safe research practices in virology.

From Fax to Secure File Transfer Protocol: The 25-Year Evolution of Real-Time Syndromic Surveillance in England.

The purpose of syndromic surveillance is to provide early warning of public health incidents, real-time situational awareness during incidents and emergencies, and reassurance of the lack of impact on the population, particularly during mass gatherings. The United Kingdom Health Security Agency (UKHSA) currently coordinates a real-time syndromic surveillance service that encompasses 6 national syndromic surveillance systems reporting on daily health care usage across England. Each working day, UKHSA analyzes syndromic data from over 200,000 daily patient encounters with the National Health Service, monitoring over 140 unique syndromic indicators, risk assessing over 50 daily statistical exceedances, and taking and recommending public health action on these daily. This English syndromic surveillance service had its origins as a small exploratory pilot in a single region of England in 1999 involving a new pilot telehealth service, initially reporting only on "cold or flu" calls. This pilot showed the value of syndromic surveillance in England, providing advanced warning of the start of seasonal influenza activity over existing laboratory-based surveillance systems. Since this initial pilot, a program of real-time syndromic surveillance has evolved from the single-system, -region, -indicator pilot (using manual data transfer methods) to an all-hazard, multisystem, automated national service. The suite of systems now monitors a wide range of syndromes, from acute respiratory illness to diarrhea to cardiac conditions, and is widely used in routine public health surveillance and for monitoring seasonal respiratory disease and incidents such as the COVID-19 pandemic. Here, we describe the 25-year evolution of the English syndromic surveillance system, focusing on the expansion and improvements in data sources and data management, the technological and digital enablers, and novel methods of data analytics and visualization.

The potential to manage releases of Bacillus anthracis using bioretention and a high flow media filter: Results of simulated runoff testing with tracer spores Bacillus globigii.

The threat of bioterrorism has spurred research on the decontamination and containment of different agents. Anthrax [causative agent Bacillus anthracis (Ba)] is a disease that can lead to severe infections within human and animals, particularly when inhaled. This research investigated the use of spore-contaminated simulated runoff events into stormwater control measures (SCMs), which are designed to retain and improve the quality of runoff and may have the potential to filter and contain the spores. In this study, the effectiveness of a bioretention cell (BRC) and high flow media filter (HFMF) in Huron, Ohio, were evaluated for removal of Bacillus globigii (Bg) spores (a harmless cognate of Ba). Three 4-8 mm simulated runoff events were created for each SCM using a fire hydrant and Bg spores were injected into the runoff upstream of the SCM inlets. The BRC significantly (p < 0.001) outperformed the HFMF in reducing Bg concentrations and loads, with an average load reduction of 1.9 log (∼99% reduction) compared to 0.4 (∼60% reduction), respectively. A probable critical design factor leading to these differences was the infiltration rate of the media and subsequent retention time within the filters, which was supported by similar disparities in suspended solids reductions. Differences in spore removal may also have been due to particle size distribution of the HFMF, which was more gravelly than the bioretention cell. At 3 and 6 months after the-simulated runoff tests, soil samples taken from both SCMs, yielding detectable Bg spores within the top 15 cm of media, with increased spore concentrations where ponding occurred for longer durations during the tests. This suggests that forebays and areas near inlets may be hotspots for spore cleanup in a real-world bioterrorism incident.

Pattern of bioterrorism in ancient times: lessons to be learned from the microbial and toxicological aspects.

The current research aimed to analyze the history of bioterrorism in antiquity and to adapt the data to modern medical knowledge. To this end, a thorough evaluation of the literature related to the ancient history of bioterrorism and modern data was done using the Web of Sciences, Science Direct, Scopus, PubMed, and Google Scholar. Results showed that knowledge of bioterrorism has existed since antiquity in different civilizations. Biological and toxicological agents were used as an instrument of legal execution, as a warfare tool in battles, or to eliminate political rivals across nations. Ancient people researched bioterrorism to apply it against enemies and at the same time provide countermeasures in favor of themselves and allies. Despite the existence of the principles of bioterrorism since ancient times, adaptation of the data to modern research can assist in planning countermeasure efforts, preventive actions, and treatments in the framework of modern counterterrorism medicine.

[Understanding public health nurses' experience, knowledge, and perception of bioterrorism training in Japan].

Objectives　Bioterrorism is a disease in which the attacks can be covert and latent, take time to manifest, and, when discovered, can result in large outbreaks. When detected, they can become large-scale outbreaks. Therefore, preparedness is essential for early detection and response for damage mitigation. Although public health nurses (PHNs) of public health centers are responsible for detecting and dealing with bioterrorism, their preparation status is not clear. Therefore, this study aimed to elucidate the status of training experience, knowledge, and awareness of bioterrorism among PHNs working in public health centers' infectious disease control departments in Japan's metropolitan areas.Methods　This was a cross-sectional, descriptive study. The participants were PHNs in the infectious disease control departments of 88 public health centers in Tokyo and its three neighboring prefectures. This are densely populated metropolitan areas with many mass gathering events and a high probability of bioterrorism. An anonymous, self-administered questionnaire survey was conducted by mail in 2019. Two PHNs per location were surveyed regarding their attributes, training experience, knowledge, and perceptions.Results　We received 71 responses (40.3%). A total of 10 (14.1%) PHNs reported having experience with bioterrorism training in the workplace. Regarding the knowledge of the four infectious diseases with a high probability of bioterrorism, more than 95% of the respondents answered they had heard of them. However, few were aware of related treatments or terrorist responses. Although survey respondents were aware of the seriousness of bioterrorism and the effectiveness of preparedness, they did not have sufficient opportunity or time to have training. They tended to be less confident in their response and were willing to have training.Conclusion　PHNs who responded to this survey do not have sufficient training experience, knowledge, opportunities, and time for bioterrorism training, and establishing a training system to improve bioterrorism preparedness is a challenge. It will be necessary for PHN students to learn about the existence of bioterrorism. Further, PHNs in public health centers need to be trained at least once using online services, including brushing up on information as needed. Based on the lessons learned from the coronavirus disease 2019 pandemic, health centers will formulate a Health Crisis Management Plan. This will be effective in fiscal 2024, and we believe that it is essential that preparedness against bioterrorism be a part of this plan.

Epidemiology of Pathogens Listed as Potential Bioterrorism Agents, the Netherlands, 2009‒2019.

We provide incidences (cases/10 million persons) in the Netherlands during 2009-2019 for pathogens listed as potential bioterrorism agents. We included pathogens from the highest categories of the European Medicines Agency or the US Centers for Disease Control and Prevention. Notifiable diseases and recently published data were used to calculate the average annual incidence. Coxiella burnetii had the highest incidence because of a Q fever epidemic during 2007-2010. Incidence then decreased to 10.8 cases/. Pathogens with an incidence >1 were Brucella spp. (2.5 cases), Francisella tularensis (1.3 cases), and Burkholderia pseudomallei (1.1 cases). Pathogens with an incidence <1 were hemorrhagic fever viruses (0.3 cases), Clostridium botulinum (0.2 cases), and Bacillus anthracis (0.1 cases). Variola major and Yersinia pestis were absent. The generally low incidences make it unlikely that ill-meaning persons can isolate these pathogens from natural sources in the Netherlands. However, the pathogens are stored in laboratories, underscoring the need for biosecurity measures.

Interventions to Reduce Risk for Pathogen Spillover and Early Disease Spread to Prevent Outbreaks, Epidemics, and Pandemics.

The pathogens that cause most emerging infectious diseases in humans originate in animals, particularly wildlife, and then spill over into humans. The accelerating frequency with which humans and domestic animals encounter wildlife because of activities such as land-use change, animal husbandry, and markets and trade in live wildlife has created growing opportunities for pathogen spillover. The risk of pathogen spillover and early disease spread among domestic animals and humans, however, can be reduced by stopping the clearing and degradation of tropical and subtropical forests, improving health and economic security of communities living in emerging infectious disease hotspots, enhancing biosecurity in animal husbandry, shutting down or strictly regulating wildlife markets and trade, and expanding pathogen surveillance. We summarize expert opinions on how to implement these goals to prevent outbreaks, epidemics, and pandemics.

Global Veterinary Diagnostic Laboratory Equipment Management and Sustainability and Implications for Pandemic Preparedness Priorities1.

Substantial investments into laboratories, notably sophisticated equipment, have been made over time to detect emerging diseases close to their source. Diagnostic capacity has expanded as a result, but challenges have emerged. The Equipment Management and Sustainability Survey was sent to the Veterinary Services of 182 countries in mid-2019. We measured the status of forty types of laboratory equipment used in veterinary diagnostic laboratories. Of the 68,455 items reported from 227 laboratories in 136 countries, 22% (14,894/68,455) were improperly maintained, and 46% (29,957/65,490) were improperly calibrated. Notable differences were observed across World Bank income levels and regions, raising concerns about equipment reliability and the results they produce. Our results will advise partners and donors on how best to support low-resource veterinary laboratories to improve sustainability and fulfill their mandate toward pandemic prevention and preparedness, as well as encourage equipment manufacturers to spur innovation and develop more sustainable products that meet end-users' needs.

Biodefense: Expanding Nursing Strategies After the SARS-CoV-2 Threat.

The COVID-19 pandemic has impacted the nursing profession and its existence in terms of preventing infection from spreading at the levels of patient care and management. Vigilance is essential in combating potential re-emerging diseases in the future. Hence, exploring a new framework, biodefense, is the best way to reframe nursing preparedness for new biological threats or new pandemics at any level of nursing care.

Systematic Review of In Vitro Antimicrobial Susceptibility Testing for Bacillus anthracis, 1947-2019.

Bacillus anthracis, the causative agent of anthrax, is a high-consequence bacterial pathogen that occurs naturally in many parts of the world and is considered an agent of biowarfare or bioterrorism. Understanding antimicrobial susceptibility profiles of B. anthracis isolates is foundational to treating naturally occurring outbreaks and to public health preparedness in the event of an intentional release. In this systematic review, we searched the peer-reviewed literature for all publications detailing antimicrobial susceptibility testing of B. anthracis. Within the set of discovered articles, we collated a subset of publications detailing susceptibility testing that followed standardized protocols for Food and Drug Administration-approved, commercially available antimicrobials. We analyzed the findings from the discovered articles, including the reported minimal inhibitory concentrations. Across the literature, most B. anthracis isolates were reported as susceptible to current first-line antimicrobials recommended for postexposure prophylaxis and treatment. The data presented for potential alternative antimicrobials will be of use if significant resistance to first-line antimicrobials arises, the strain is bioengineered, or first-line antimicrobials are not tolerated or available.

Epidemiologic Features and Control Measures during Monkeypox Outbreak, Spain, June 2022.

During June 2022, Spain was one of the countries most affected worldwide by a multicountry monkeypox outbreak with chains of transmission without identified links to disease-endemic countries. We provide epidemiologic features of cases reported in Spain and the coordinated measures taken to respond to this outbreak.

Laboratory Misidentifications Resulting from Taxonomic Changes to Bacillus cereus Group Species, 2018-2022.

Whole-genome sequencing (WGS) is being applied increasingly to Bacillus cereus group species; however, misinterpretation of WGS results may have severe consequences. We report 3 cases, 1 of which was an outbreak, in which misinterpretation of B. cereus group WGS results hindered communication within public health and industrial laboratories.

Bioterrorism: An analysis of biological agents used in terrorist events.

BACKGROUND: The Covid19 pandemic has reignited debates and discussions around healthcare systems' biosecurity vulnerabilities and cast a spotlight on the potential weaponization of biological agents. Terrorist and violent extremist groups have already attempted to incite the intentional spread of Covid19 and to use it as an improvised form of a biological weapon. This study aims to provide an epidemiological description of all terrorism-related attacks using biological agents sustained between 1970 and 2019. METHODS: Data collection was performed using a retrospective database search through the Global Terrorism Database (GTD). The GTD was searched using the internal database search functions for all events using biological weapons between January 1, 1970 - December 31, 2019. RESULTS: 33 terrorist attacks involving biological agents were recorded between 1970 and 2019, registering 9 deaths and 806 injuries. 21 events occurred in the United States, 3 in Kenya, 2 each in both the United Kingdom and Pakistan and a single event in Japan, Columbia, Israel, Russia and Tunisia. CONCLUSION: The reported use of biological agents as a terrorist weapon is extremely rare and accounts for 0.02% of all historic terrorist attacks. Despite its apparent rarity, however, bioterrorism has the ability to inflict mass injuries unmatched by conventional weapons. Anthrax has been the most commonly used in previous bioterrorism events with the vast majority of reported attacks occurring in the United States by a single suspected perpetrator. Counter-Terrorism Medicine (CTM) and Disaster Medicine (DM) specialists need to be proactive in delivering ongoing educational sessions on biological events to first responder communities, and anticipate emerging novel biotechnology threats.

Knowledge and practice of nursing students regarding bioterrorism and emergency preparedness: comparison of the effects of simulations and workshop.

BACKGROUND AND PURPOSE: Bioterrorism is a global threat. Nurses are one of the first groups that should be ready for it. College years are the best time to educate these issues. This study aimed to compare the effectiveness of simulation and workshop on knowledge and practice of nursing students regarding bioterrorism. MATERIALS AND METHODS: This was an experimental study. The study sample consisted of 40 last-year nursing students who were randomly assigned to two groups by using random numbers table. Data was collected using a demographic questionnaire, bioterrorism knowledge scale, and an OSCE checklist. Before the intervention, the students completed the study questionnaires and a six-station OSCE test. The workshop group (20 students) learned how to deal with bioterrorism through lectures. The simulation group (20 students) participated in a simulation learning program. After one month, the students completed the study tools again. Finally, collected data were analyzed using descriptive and inferential statistics in SPSS V.16. RESULTS: The difference between the knowledge and performance scores of both groups (workshop and simulation), before and after the intervention, was statistically significant (P < 0.001). Students in both groups had higher knowledge and performance scores after the intervention. The simulation group scores were higher than the workshop group scores in the knowledge and the most of performance domains. CONCLUSION: The simulation group had better results in terms of enhancing knowledge, preparedness, disaster triage, reporting, incident management, communication, mental disorders, and isolation domains compared to the workshop group.

Dental professionals' attitude and willingness to respond during disaster events: a cross-sectional study.

INTRODUCTION: In the present times, natural and man-made threats have questioned our existence on this planet. Health care professionals need to be aware of all the procedures to follow during such an event that threatens to paralyze the entire community and should be able to respond effectively. OBJECTIVE: The present study was conducted to assess the willingness and attitude of dental professionals to render help during any disaster or catastrophic event. MATERIAL AND METHODS: The present cross-sectional study among 480 subjects who were residing in Tricity (Chandigarh, Mohali and Panchkula). Informed consent was taken from all the subjects. Systematic random sampling methodology was employed for selection for study sample. The study utilized a self-designed close-ended questionnaire written in English and verified by experts. The questionnaire was divided into two parts to collect the required information. The data was analyzed using SPSS software version 21; chi-square test and multiple regression analysis were used to arrive at the results. RESULTS: More than 85% of subjects had awareness regarding natural and manmade disasters. A positive response was given by 79.2% of subjects regarding willingness to help during disaster. Educational qualification of subjects was significantly associated with attitude towards some aspects of disaster response (p<0.05). Postgraduate subjects and subjects who were academicians were more willing to render help (OR: 2.18 & 3.65 respectively). CONCLUSIONS: The study emphasizes the need for educational and training programs for dentists' regarding disaster management. A short course on emergency and disaster management can be included in undergraduate and postgraduate dental curriculum.