Animal Biosafety Level Research Articles

Some Specific Biological Risks of SARS-CoV-2 Infected Syrian Hamsters at Animal Biosafety Level - Lab 3

The emergence of novel severe acute respiratory syndrome 2 (SARS-CoV-2) has prompted extensive research to identify its pathogenesis, transmission, and reservoir entry in animals. Syrian hamsters (Mesocricetus auratus) are being identified as an ideal animal model for the study of SARS-CoV-2 infection due to their susceptibility and respiratory symptoms similar to that observed in humans. This study focused on the specific biological risks associated with the use of infected Syrian hamsters in animal biosafety level 3 (ABSL-3) laboratories. The professional and psychological training of professionals working with laboratory animals should be a priority. Biological safety requires the involvement of all parties from technical staff to top management. The COVID-19 pandemic has shown a limitation in experimental studies when obtaining the right strain of mice proved difficult. A rare experimental model of Syrian hamsters started to be used in the work, which Syrian hamsters have several distinctive characteristics associated with increased aggressiveness of females, leading to a requirement to keep them isolated and some technical difficulties during manipulations in the biological safety cabinet. Therefore, a risk-based approach must be introduced to reduce the additional risks to workers to an acceptable level. This includes enhanced accuracy in the use of personal protective equipment (PPE), rigorously following biosecurity protocols, and monitoring staff health to detect potential risk of zoonotic transmission. The implementation of international standards and regulations on biological safety and biosecurity is of paramount importance.

Evaluation of Whole Carcass Composting as a Mortality Disposal Option for African Swine Fever Virus-Infected Swine

African swine fever (ASF) epizootic outbreaks often result in the deaths of large numbers of swine either through confirmed or suspected disease or depopulation for outbreak control. Identification of effective biosecure livestock mortality disposal methods, capable of inactivating ASF virus (ASFV) in contaminated carcasses, remains an important component for development of localized outbreak response and recovery plans. In this study, we evaluated ASFV inactivation and viral DNA degradation during composting of ASFV-infected swine carcasses in an Animal Biosafety Level 3-Ag (ABSL3-Ag) high-containment vivarium utilizing virus isolation (VI) and real-time polymerase chain reaction (RT-PCR). Four ASFV-infected pigs, approximately 54 kg, were composted for 37 days in a static, nonaerated windrow assembled using wood chips, horse manure, and pine shavings as the primary sources of carbon. The windrow daily temperature was sustained at ≥55°C for 18 days. Spleen samples, collected over 28 days, were negative for infectious ASFV by virus isolation on day 5 but remained positive for ASFV DNA by RT-PCR through day 28. At the conclusion of the study, bone marrow, muscle, and pinna samples were collected from composted carcasses, in addition to carbon materials in the windrow. No infectious ASFV was detected by virus isolation in any of the decomposed tissues or carbon samples. However, ASFV DNA remained detectable in all swine tissues (n = 36) and in 14/16 sentinel carbon samples at the end of the study, indicating that positive detection of ASFV DNA did not correlate with virus infectivity. In conclusion, infectious ASFV was rapidly eliminated in ASFV-infected swine during composting of whole carcasses by day 5, coinciding with a temperature increase in the compost pile to ≥55°C.

Computed Tomography Imaging for Monitoring of Marburg Virus Disease: a Nonhuman Primate Proof-Of-Concept Study.

Marburg virus (MARV) is a highly virulent zoonotic filovirid that causes Marburg virus disease (MVD) in humans. The pathogenesis of MVD remains poorly understood, partially due to the low number of cases that can be studied, the absence of state-of-the-art medical equipment in areas where cases are reported, and limitations on the number of animals that can be safely used in experimental studies under maximum containment animal biosafety level 4 conditions. Medical imaging modalities, such as whole-body computed tomography (CT), may help to describe disease progression in vivo, potentially replacing ethically contentious and logistically challenging serial euthanasia studies. Towards this vision, we performed a pilot study, during which we acquired whole-body CT images of 6 rhesus monkeys before and 7 to 9 days after intramuscular MARV exposure. We identified imaging abnormalities in the liver, spleen, and axillary lymph nodes that corresponded to clinical, virological, and gross pathological hallmarks of MVD in this animal model. Quantitative image analysis indicated hepatomegaly with a significant reduction in organ density (indicating fatty infiltration of the liver), splenomegaly, and edema that corresponded with gross pathological and histopathological findings. Our results indicated that CT imaging could be used to verify and quantify typical MVD pathogenesis versus altered, diminished, or absent disease severity or progression in the presence of candidate medical countermeasures, thus possibly reducing the number of animals needed and eliminating serial euthanasia. IMPORTANCE Marburg virus (MARV) is a highly virulent zoonotic filovirid that causes Marburg virus disease (MVD) in humans. Much is unknown about disease progression and, thus, prevention and treatment options are limited. Medical imaging modalities, such as whole-body computed tomography (CT), have the potential to improve understanding of MVD pathogenesis. Our study used CT to identify abnormalities in the liver, spleen, and axillary lymph nodes that corresponded to known clinical signs of MVD in this animal model. Our results indicated that CT imaging and analyses could be used to elucidate pathogenesis and possibly assess the efficacy of candidate treatments.

Establishment of multicenter COVID-19 therapeutics preclinical test system in Republic of Korea

Throughout the recent COVID-19 pandemic, South Korea led national efforts to develop vaccines and therapeutics for SARS-CoV-2. The project proceeded as follows: 1) evaluation system setup (including Animal Biosafety Level 3 (ABSL3) facility alliance, standardized nonclinical evaluation protocol, and laboratory information management system), 2) application (including committee review and selection), and 3) evaluation (including expert judgment and reporting). After receiving 101 applications, the selection committee reviewed pharmacokinetics, toxicity, and efficacy data and selected 32 final candidates. In the nonclinical efficacy test, we used golden Syrian hamsters and human angiotensin-converting enzyme 2 transgenic mice under a cytokeratin 18 promoter to evaluate mortality, clinical signs, body weight, viral titer, neutralizing antibody presence, and histopathology. These data indicated eight new drugs and one repositioned drug having significant efficacy for COVID-19. Three vaccine and four antiviral drugs exerted significant protective activities against SARS-CoV-2 pathogenesis. Additionally, two anti-inflammatory drugs showed therapeutic effects on lung lesions and weight loss through their mechanism of action but did not affect viral replication. Along with systematic verification of COVID-19 animal models through large-scale studies, our findings suggest that ABSL3 multicenter alliance and nonclinical evaluation protocol standardization can promote reliable efficacy testing against COVID-19, thus expediting medical product development.

Gut microbiota modulates lung fibrosis severity following acute lung injury in mice

Independent studies demonstrate the significance of gut microbiota on the pathogenesis of chronic lung diseases; yet little is known regarding the role of the gut microbiota in lung fibrosis progression. Here we show, using the bleomycin murine model to quantify lung fibrosis in C57BL/6 J mice housed in germ-free, animal biosafety level 1 (ABSL-1), or animal biosafety level 2 (ABSL-2) environments, that germ-free mice are protected from lung fibrosis, while ABSL-1 and ABSL-2 mice develop mild and severe lung fibrosis, respectively. Metagenomic analysis reveals no notable distinctions between ABSL-1 and ABSL-2 lung microbiota, whereas greater microbial diversity, with increased Bifidobacterium and Lactobacilli, is present in ABSL-1 compared to ABSL-2 gut microbiota. Flow cytometric analysis reveals enhanced IL-6/STAT3/IL-17A signaling in pulmonary CD4 + T cells of ABSL-2 mice. Fecal transplantation of ABSL-2 stool into germ-free mice recapitulated more severe fibrosis than transplantation of ABSL-1 stool. Lactobacilli supernatant reduces collagen 1 A production in IL-17A- and TGFβ1-stimulated human lung fibroblasts. These findings support a functional role of the gut microbiota in augmenting lung fibrosis severity.

Infection Characteristics and Transcriptomics of African Swine Fever Virus in Bama Minipigs.

Animal experiments on African swine fever virus (ASFV) are vital to the study of ASFV; however, ASFV can only infect pigs, and animal experiments need to be performed in animal biosafety level 3 (ABSL-3) laboratories, meaning that many small ABSL-3 laboratories are unable to carry out in vivo ASFV experiments. Therefore, miniaturized experimental animals for ASFV infection are urgently needed. Here, we successfully isolated genotype II of ASFV SY-1 from wild boars and evaluated ASFV-infected Bama minipigs in a negative-pressure isolator of a small ABSL-3 laboratory. The pathological changes of ASFV-infected Bama minipigs were consistent with characteristic lesions of ASFV-infected domestic pigs and wild boars. All pigs died 5 to 14 days postinfection (dpi) through intramuscular injection. Viral genomic DNA from nasal, oral, and rectal swab samples was first detectable at 2 to 4 dpi. The common differentially expressed genes were clustered in the immune-related, metabolic, and inflammatory response pathways from the spleen and inguinal lymph node samples comparing infected to mock. In summary, these results demonstrated that the Bama minipig was an appropriate model for ASFV infection in small ABSL-3 laboratories that can accelerate the research of vaccines and antiviral drugs and uncover pathogenic mechanisms of ASFV infection. IMPORTANCE African swine fever virus (ASFV) can only infect pigs rather than other animals. However, the domestic pigs cannot be kept in small ABSL-3 laboratories for a long time due to the characteristics of rapid growth and large size, which hinder ASFV research, including research of vaccines, antiviral drugs, and mechanisms. In contrast, Bama minipigs have unique advantages consisting of low growth and small size. In the research, Bama minipigs were used to evaluate the characteristics of ASFV infection in small ABSL-3 laboratories. The pathological changes, viral shedding, and gene regulation were consistent with those of domestic pigs infected with ASFV. Therefore, Bama minipigs can be a suitable model for ASFV infection in small ABSL-3 laboratories.

Persistent intraocular Ebola virus RNA is associated with severe uveitis in a convalescent rhesus monkey

Despite increasing evidence that uveitis is common and consequential in survivors of Ebola virus disease (EVD), the host-pathogen determinants of the clinical phenotype are undefined, including the pathogenetic role of persistent viral antigen, ocular tissue-specific immune responses, and histopathologic characterization. Absent sampling of human intraocular fluids and tissues, these questions might be investigated in animal models of disease; however, challenges intrinsic to the nonhuman primate model and the animal biosafety level 4 setting have historically limited inquiry. In a rhesus monkey survivor of experimental Ebola virus (EBOV) infection, we observed and documented the clinical, virologic, immunologic, and histopathologic features of severe uveitis. Here we show the clinical natural history, resultant ocular pathology, intraocular antigen-specific antibody detection, and persistent intraocular EBOV RNA detected long after clinical resolution. The association of persistent EBOV RNA as a potential driver of severe immunopathology has pathophysiologic implications for understanding, preventing, and mitigating vision-threatening uveitis in EVD survivors.

Experimentally controlled study indicates that the naturally occurring recombinant vaccine-like lumpy skin disease strain Udmurtiya/2019, detected during freezing winter in northern latitudes, is transmitted via indirect contact.

Lumpy skin disease (LSD) caused by LSD virus (LSDV), is a member of the poxvirus genus Capripoxvirus. It is classified as a notifiable disease by the World Organization for Animal Health (WOAH) based on its potential for rapid spread and global economic impact. Due to these characteristics, the mode of LSDV transmission has prompted intensive research efforts. Previous experimental studies using the virulent vaccine-derived recombinant LSDV strain Saratov/2017, demonstrated that this strain has the capacity for transmission in a vector-proof environment. This study demonstrated that a second novel recombinant vaccine-derived LSDV strain Udmurtiya/2019, can infect bulls in contact with diseased animals, in the absence of insect vectors. Bulls were housed in an insect proof animal biosafety level 3 facility, where half the animals were inoculated intravenously with the recombinant LSDV (Udmurtiya/2019), whilst the remaining five animals were mock-inoculated but kept in contact with the inoculated group. Both the infected / inoculated group (IN) and uninfected / incontact group (IC), were monitored for 41 days with continuous registration of body temperature, observations for clinical signs and collection of blood samples and nasal swabs for testing of LSDV presence using real-time PCR. Results indicated that cohabitation of animals from both groups was sufficient to transmit the virus from the IN to the IC-group, with the onset of clinical signs including pyrexia (~41°C) and classical LSD nodular skin lesions starting at 10 dpi for the IN group and 16 dpi for the IC-group. Additionally, the presence of LSDV genomes as well as anti-LSDV antibodies were detected in swabs, blood and serum samples from animals belonging to both groups. These results provides additional evidence of LSDV transmission in a controlled environment without direct contact between diseased and healthy animals, yet in the absence of vectors. Based on these observations, the question concerning a hypothetical relation between mutations in the virus genome and its mode of transmission gains more importance and requires additional investigations with direct comparisons between classical and novel recombinant LSDV strains.

Laboratory information management system for COVID-19 non-clinical efficacy trial data

BackgroundAs the number of large-scale studies involving multiple organizations producing data has steadily increased, an integrated system for a common interoperable format is needed. In response to the coronavirus disease 2019 (COVID-19) pandemic, a number of global efforts are underway to develop vaccines and therapeutics. We are therefore observing an explosion in the proliferation of COVID-19 data, and interoperability is highly requested in multiple institutions participating simultaneously in COVID-19 pandemic research.ResultsIn this study, a laboratory information management system (LIMS) approach has been adopted to systemically manage various COVID-19 non-clinical trial data, including mortality, clinical signs, body weight, body temperature, organ weights, viral titer (viral replication and viral RNA), and multiorgan histopathology, from multiple institutions based on a web interface. The main aim of the implemented system is to integrate, standardize, and organize data collected from laboratories in multiple institutes for COVID-19 non-clinical efficacy testings. Six animal biosafety level 3 institutions proved the feasibility of our system. Substantial benefits were shown by maximizing collaborative high-quality non-clinical research.ConclusionsThis LIMS platform can be used for future outbreaks, leading to accelerated medical product development through the systematic management of extensive data from non-clinical animal studies.

It's the Wrong Virus: Rapid Adaptation of Operations Inside a Highly Pathogenic Avian Influenza Select Agent Laboratory in Response to the 2019 SARS-CoV-2 Pandemic.

The Animal Biosafety Level 3 Enhanced (ABSL-3+) laboratory at St. Jude Children's Research Hospital has a long history of influenza pandemic preparedness. The emergence of SARS-CoV-2 and subsequent expansion into a pandemic has put new and unanticipated demands on laboratory operations since April 2020. Administrative changes, investigative methods requiring increased demand for inactivation and validation of sample removal, and the adoption of a new animal model into the space required all arms of our Biorisk Management System (BMS) to respond with speed and innovation. In this report, we describe the outcomes of three major operational changes that were implemented to adapt the ABSL-3+ select agent space into a multipathogen laboratory. First were administrative controls that were revised and developed with new Institutional Biosafety Committee protocols, laboratory space segregation, training of staff, and occupational health changes for potential exposure to SARS-CoV-2 inside the laboratory. Second were extensive inactivation and validation experiments performed for both highly pathogenic avian influenza and SARS-CoV-2 to meet the demands for sample removal to a lower biosafety level. Third was the establishment of a new caging system to house Syrian Golden hamsters for SARS-CoV-2 risk assessment modeling. The demands placed on biocontainment laboratories for response to SARS-CoV-2 has highlighted the importance of a robust BMS. In a relatively short time, the ABSL-3+ was able to adapt from a single select agent space to a multipathogen laboratory and expand our pandemic response capacity.

Results of a 2020 Survey on Reporting Requirements and Practices for Biocontainment Laboratory Accidents.

Biosafety laboratory accidents are a normal part of laboratory science, but the frequency of such accidents is unclear due to current reporting standards and processes. To better understand accident reporting, a survey was created, with input from ABSA International, which included a series of questions about standards, requirements, and likely motivations for reporting or nonreporting. A total of 60 biosafety officers completed the survey. Respondents reported working with more than 5,000 people in laboratories, including more than 40 biosafety level 3 or animal biosafety level 3 laboratories, which work with higher-risk pathogens. Most of the respondents were located in the United States, Canada, or New Zealand, or did not identify their location. Notable results included that 97% of surveyed biosafety officers oversee laboratories that require reporting exposure to at least some pathogens. However, 63% relayed that the reports are not usually sent outside of the institution where they occurred. A slight majority (55%) stated that paper reports were used, with the rest reporting they used a variety of computer systems. Even in laboratories that used paper-based reporting systems, 67% relayed that these reports were used alongside, or entered into, a digital system. While 82% of these biosafety officers agreed that workers understood the importance of reporting for their own safety, 82% also agreed that a variety of disincentives prevent laboratory workers from reporting incidents, including concerns about job loss and loss of funding.

Analysis of Range and Use of a Hybrid Hydrogen Peroxide System for Biosafety Level 3 and Animal Biosafety Level 3 Agriculture Laboratory Decontamination.

Introduction:The applications of fumigation and the challenges that high-containment facilities face in achieving effective large volume decontamination are well understood. The Biosecurity Research Institute at Kansas State University sought to evaluate a novel system within their biosafety level 3 (BSL-3) and animal biosafety level 3 agriculture (ABSL-3Ag) facility.Methods:The system chosen for this study is the CURIS® Hybrid Hydrogen PeroxideTM (HHPTM) system, comprising a mobile 36-pound (16 kg) device delivering a proprietary 7% hydrogen peroxide (H2O2) solution. To examine the system's efficacy in multiple laboratory settings, two BSL-3 laboratories (2,281 [65 m3] and 4,668 ft3 [132 m3]) with dropped ceiling interstitial spaces and an ABSL-3Ag necropsy suite (44,212 ft3 [1,252 m3]) with 21-foot (6.4 m) ceilings were selected. Biological indicators (BIs) of Geobacillus stearothermophilus (1.7 × 106 organisms) on steel spore carriers and H2O2 chemical indicators (CIs) were used to provide validation.Results:After cycle optimization, the smaller laboratory had a total of 60 BIs over two treatments that demonstrated a greater than 6-log reduction of bacterial spores. The larger laboratory (192 BIs) and the necropsy suite (206 BIs) had no BIs positive for spore growth when incubated at 60°C for 24 h per manufacturer's specifications.Conclusion:Overall successful results through multiple components of this study demonstrate that the HHP device, paired with the pulsed 7% H2O2 solution, achieved efficacy regardless of variables in laboratory size and layout. Perceived challenges such as 21-ft (6.4 m) ceiling heights, active equipment, and difficult to access ceiling interstitial spaces proved unfounded. Given the successful sterilization of all challenged BIs, the HHP system presents a useful alternative for high level decontamination within BSL-3 and ABSL-3Ag facilities.

Significance of High-Containment Biological Laboratories Performing Work During the COVID-19 Pandemic: Biosafety Level-3 and -4 Labs.

High containment biological laboratories (HCBL) are required for work on Risk Group 3 and 4 agents across the spectrum of basic, applied, and translational research. These laboratories include biosafety level (BSL)-3, BSL-4, animal BSL (ABSL)-3, BSL-3-Ag (agriculture livestock), and ABSL-4 laboratories. While SARS-CoV-2 is classified as a Risk Group 3 biological agent, routine diagnostic can be handled at BSL-2. Scenarios involving virus culture, potential exposure to aerosols, divergent high transmissible variants, and zoonosis from laboratory animals require higher BSL-3 measures. Establishing HCBLs especially those at BSL-4 is costly and needs continual investments of resources and funding to sustain labor, equipment, infrastructure, certifications, and operational needs. There are now over 50 BSL-4 laboratories and numerous BSL-3 laboratories worldwide. Besides technical and funding challenges, there are biosecurity and dual-use risks, and local community issues to contend with in order to sustain operations. Here, we describe case histories for distinct HCBLs: representative national centers for diagnostic and reference, nonprofit organizations. Case histories describe capabilities and assess activities during COVID-19 and include capacities, gaps, successes, and summary of lessons learned for future practice.

Natural history of disease in cynomolgus monkeys exposed to Ebola virus Kikwit strain demonstrates the reliability of this non-human primate model for Ebola virus disease.

Filoviruses (Family Filoviridae genera Ebolavirus and Marburgvirus) are negative-stranded RNA viruses that cause severe health effects in humans and non-human primates, including death. Except in outbreak settings, vaccines and other medical countermeasures against Ebola virus (EBOV) will require testing under the FDA Animal Rule. Multiple vaccine candidates have been evaluated using cynomolgus monkeys (CM) exposed to EBOV Kikwit strain. To the best of our knowledge, however, animal model development data supporting the use of CM in vaccine research have not been submitted to the FDA. This study describes a large CM database (122 CM, 62 female and 60 male, age 2 to 9 years) and demonstrates the consistency of the CM model through time to death models and descriptive statistics. CMs were exposed to EBOV doses of 0.1 to 100,000 PFU in 33 studies conducted at three Animal Biosafety Level 4 facilities, by three exposure routes. Time to death was modeled using Cox proportional hazards models with a frailty term that incorporated study-to-study variability. Despite significant differences attributed to exposure variables, all CMs exposed to the 100 to 1,000 pfu doses commonly used in vaccine studies died or met euthanasia criteria within 21 days of exposure, median 7 days, 93% between 5 and 12 days of exposure. Moderate clinical signs were observed 4 to 5 days after exposure and preceded death or euthanasia by approximately one day. Viremia was detected within a few days of infection. Hematology indices were indicative of viremia and the propensity for hemorrhage with progression of Ebola viremia. Changes associated with coagulation parameters and platelets were consistent with coagulation disruption. Changes in leukocyte profiles were indicative of an acute inflammatory response. Increased liver enzymes were observed shortly after exposure. Taken together, these factors suggest that the cynomolgus monkey is a reliable animal model for human disease.

Outside the Box: Working With Wildlife in Biocontainment.

Research with captive wildlife in Animal Biosafety Level 2 (ABSL2) and 3 (ABSL3) facilities is becoming increasingly necessary as emerging and re-emerging diseases involving wildlife have increasing impacts on human, animal, and environmental health. Utilizing wildlife species in a research facility often requires outside the box thinking with specialized knowledge, practices, facilities, and equipment. The USGS National Wildlife Health Center (NWHC) houses an ABSL3 facility dedicated to understanding wildlife diseases and developing tools to mitigate their impacts on animal and human health. This review presents considerations for utilizing captive wildlife for infectious disease studies, including, husbandry, animal welfare, veterinary care, and biosafety. Examples are drawn from primary literature review and collective 40-year experience of the NWHC. Working with wildlife in ABSL2 and ABSL3 facilities differs from laboratory animals in that typical laboratory housing systems, husbandry practices, and biosafety practices are not designed for work with wildlife. This requires thoughtful adaptation of standard equipment and practices, invention of customized solutions and development of appropriate enrichment plans using the natural history of the species and the microbiological characteristics of introduced and native pathogens. Ultimately, this task requires critical risk assessment, understanding of the physical and psychological needs of diverse species, creativity, innovation, and flexibility. Finally, continual reassessment and improvement are imperative in this constantly changing specialty area of infectious disease and environmental hazard research.

IACUC and Veterinary Considerations for Review of ABSL3 and ABSL4 Research Protocols.

With the recent upswing of infectious disease outbreaks (coronavirus, influenza, Ebola, etc), there is an ever-increasing need for biocontainment animal use protocols to better address the research of emerging diseases and to increase the health of both animals and humans. It is imperative that we as a research community ensure these protocols are conducted with the utmost scrutiny and regulatory compliance for the welfare of the animals as well as the health and safety concerns of the individual conducting these studies. Both the welfare of the animals and the health and safety of the research staff must be balanced with the integrity of the science being studied. Even prior to reviewing biocontainment protocols, the research stakeholders should have professional and collegial interactions across all levels of the proposed project. These stakeholders should include the attending veterinarian, the principal investigator, the sponsor, and any organic institutional health and safety assets (environmental health and safety, occupational health, biosafety personnel, medical personnel, facilities operations and maintenance, etc). At most institutions, these stakeholders are members of the Institutional Animal Care and Use Committee and may not possess the necessary tools to properly assess an Animal Biosafety Level 3 and 4 animal use protocol. It is the goal of this article to review some basic concepts of biocontainment, discuss critical communications and preapprovals, clinical observations, medical interventions and supportive care, scientific and study endpoints, euthanasia criteria, animal manipulations, documentation, training, emergency response and contingency plans, security, and decontamination and provide a scenario-based and informative thought-provoking process Institutional Animal Care and Use Committee members and veterinary staff may consider during Animal Biosafety Level 3 and 4 protocol review. These topics will enhance the ability of all stakeholders to balance the protection of the people with the integrity of the science and ultimately the welfare of the animal.

Challenges and Opportunities in the Use of High and Maximum Biocontainment Facilities in Developing and Licensing Risk Group 3 and Risk Group 4 Agent Veterinary Vaccines.

New solutions are necessary for the singular global health security threat formed by endemic, epidemic, and emerging/re-emerging zoonoses, coupled with epizootic and enzootic transboundary animal diseases (TADs). This One Health issue is related to the daily interactions between wildlife, domesticated and indigenous livestock, and humans primarily associated with global trade, transboundary co-movement of humans and diverse livestock/livestock products, and agriculture production intensification and penetration into previously uninhabited areas. The World Health Organization defines Risk Group 3 (RG-3) and RG-4 pathogens as mainly viruses but also bacteria that serve as the foundation for approximately 60% of emerging infectious diseases that are zoonoses. The World Organisation for Animal Health defines trade-notifiable TADs, and subsets of these are zoonotic. Livestock vaccination policies mainly focus on TADs that are promulgated by the United Nations Food and Agriculture Organization and government agriculture agencies. The development, licensure, and product manufacturing of next-generation molecular-based RG-3 and RG-4 veterinary vaccines largely ignored by the global animal health biopharmaceutical sector can have an important positive impact on food security and One Health. There have been sharp increases in the global demand for livestock meat and milk products, especially in low- and middle-income countries in Africa and Asia. This relatively recent market driver-coupled with scientific advances in human EID and zoonotic disease vaccine platform technologies and increases in the number of high (US biosafety level 3 agriculture) and maximum (US animal biosafety level 4) biocontainment facilities with supporting workforce capabilities-offers new investment opportunities to the animal health biopharmaceutical sector. Moreover, a growing number of One Health public-private partnerships have moved the net present value calculus in favor of the financial feasibility of RG-3 and RG-4 veterinary vaccine product development and licensure. This article highlights the challenges and opportunities in the use of high and maximum biocontainment facilities in developing and licensing RG-3 and RG-4 veterinary vaccines that are safe and effective against epizootic and enzootic TADs and zoonotic diseases.

Challenges and Solutions With Agricultural Animal High Containment Waste Disposal.

Waste disposal in Agricultural Animal High Containment Animal Biosafety Level 3Ag and Animal Biosafety Level 4Ag (ABSL-3Ag and ABSL-4Ag) research facilities necessitates significantly more attention to detail in operations than that required in lower-containment-level laboratories. The unique features and requirements of agricultural-related research involve additional equipment and systems to safely transfer decontaminated waste out of the facility. The waste stream coming from ABSL-3Ag and ABSL-4Ag facilities, or high containment agricultural research waste, consists of many forms and differs from most research facility waste in that it is produced from research with livestock or other species loose housed, with the animal room serving as primary containment. This is in contrast to small laboratory animals being housed in primary containment caging. Waste handling equipment in agricultural research facilities may include autoclaves, effluent decontamination systems, incinerators, high-temperature renderers, alkaline tissue digester systems, high-efficiency particulate air filtration of exhaust and supply air, gas decontamination systems, and laundry facilities. This article focuses primarily on the disposal of waste from ABSL-3Ag livestock facilities, including procedures and lessons learned over 10 years of facility operation.

ANSI/ASSP Z9.14-2020 Testing and Performance-Verification Methodologies for Biosafety Level 3 (BSL-3) and Animal Biosafety Level 3 (ABSL-3) Ventilation Systems.

The American National Standards Institute has approved the revised standard ANSI/ASSP Z9.14-2020, "Testing and Performance-Verification Methodologies for Biosafety Level 3 (BSL-3) and Animal Biosafety Level 3 (ABSL-3) Ventilation Systems"; the revision was published on March 31, 2020. ANSI/ASSP Z9.14-2020 focuses on performance verification of engineering controls related specifically to ventilation system features of BSL-3/ABSL-3 facilities. The revised standard has been enhanced to include new definitions; new guidance for developing corrective action plans and performing verification of conformance to regulations, appendices, risk assessment matrices, and checklists dedicated to performing a facility risk assessment; and numerous updates to the original content.

SARS-CoV-2: an unknown agent and challenges in vaccine development

SARS-CoV-2: an unknown agent and challenges in vaccine development