Search Results (217)

Background: Outer Membrane Vesicles (OMVs), nanosized particles derived from Gram-negative bacteria, are promising vaccine carriers due to innate immunogenicity and self-adjuvant properties. Yet the systematic evaluations of OMV-associated toxicity remain limited. Methods: We developed a CRISPR/Cas9-engineered Salmonella enterica serovar Typhimurium ΔmsbB mutant (Mut4_STM) to produce detoxified OMVs (Mut4_OMVs) with enhanced yield. Subcutaneous immunization of BALB/c mice with Mut4_OMVs to evaluate safety, and the adjuvant efficacy was also determined by injecting Mut4_OMVs with Yersinia pestis F1Vmut or Bacillus anthracis PA_D4 antigens, mixing formulation, respectively. Results: Mut4_OMVs showed a 9-fold protein yield increase over wild-type OMVs. While all mice injected with wild-type OMVs died, 100% survival was observed in those receiving Mut4_OMVs. However, dose-dependent pathological alterations became evident in specific organs as the administration dose escalated, such as induced splenic extramedullary hematopoiesis and renal edema. Despite residual toxicity, 2–3 doses of 10 μg Mut4_OMVs elicited antigen-specific antibody titers comparable to aluminum adjuvant controls and superior T-cell immune responses. Conclusion: While Mut4_OMVs retain potent adjuvant activity, their residual toxicity necessitates further biocompatibility optimization for safe vaccine applications. Full article

Zoonotic diseases present a growing public health challenge, particularly in sub-Saharan Africa (SSA) due to close interactions between humans and animals and poor diagnostic capacity. This pilot study investigated human exposure to zoonotic pathogens in Zambia among 47 suspected COVID-19 patients from whom nasopharyngeal samples were collected between November 2020 and February 2021 at two major COVID-19 referral centers in Lusaka. Using metagenomic next-generation sequencing (mNGS), the study identified a diverse range of pathogens, including bacterial, fungal, viral, and parasitic species. The prevalence of zoonotic pathogens was 57.4%. Noteworthy zoonoses included Bacillus anthracis, Sporothrix schenckii, Listeria monocytogenes, Yersinia pestis, Streptococcus suis, Vibrio parahaemolyticus, Brucella melitensis, Rickettsia prowazekii, Shewanella algae, Rickettsia japonica, Coxiella burnetii, Leptospira borgpetersenii, Erysipelothrix rhusiopathiae, Brucella abortus, Bartonella quintana, Banna virus, Vibrio alginolyticus, Bartonella clarridgeiae, Rickettsia canadensis, Leishmania braziliensis, Trypanosoma brucei, Pasteurella multocida, and Arcobacter butzleri. Despite moderate diversity in the microbial community, no significant demographic or health-related factors, including age, gender, or comorbidities such as HIV, were found to be statistically associated with zoonotic pathogen infection. The findings provide valuable data on the presence of zoonotic pathogens in humans in Zambia and highlight the need for more comprehensive research into zoonotic diseases in both clinical and non-clinical settings. Full article

In developing countries like Bangladesh, livestock is one of the main sources of income. Among several infectious diseases, the Gram-positive bacterium Bacillus anthracis causes a zoonotic disease named anthrax. Animal anthrax outbreaks are a frequently occurring problem in Bangladesh. Our present study aims to molecularly identify and characterize B. anthracis from three districts of Bangladesh by 16S rRNA gene sequencing. B. anthracis was confirmed in soil, meat, and blood samples using PCR. Anthrax-affected soil (n = 128), blood (n = 1), and meat (n = 2) samples were analyzed using PCR. One of the positive samples was randomly chosen for sequencing, and MEGA5 software was used to generate the phylogenetic tree from the sequencing result. A total of 21 (16.40%) soil samples and all of the blood and meat samples were positive for the presence of bacteria, confirmed by PCR. The 16S rRNA gene of B. anthracis Sirajganj-1 was identical to that of other strains. To fulfill the Sustainable Development Goals, it is important to control zoonotic diseases. Our results may help discover the virulent genes of B. anthracis for future investigation and control this zoonotic disease. Also, a proper awareness of vaccination and effective surveillance system is important to eradicate any kind of zoonotic disease in developing nations. Full article

The bacterium Bacillus anthracis secretes three protein exotoxins: Protective Antigen 83 (PA83), Lethal Factor (LF), and Edema Factor (EF). A cleaved form of PA83 (PA63) aids LF and EF entry into the cytoplasm, which leads to anthrax-induced cell death. The Protein Data Bank (PDB) has more than 25 structures of LF: the monomer alone, bound with inhibitors, or bound to PA63. The structures are all—with only minor shifts of a few Ångströms—nearly congruent. We have measured the structure of LF at equilibrium in D₂O solution by small-angle neutron scattering (SANS). The shape is modeled well by a parallelepiped (all angles 90°) with dimensions of 12 Å × 49 Å × 129 Å. For a protein with a typical density of 1.4, the molecular weight would be between 55 and 94 kDa, which is comparable to that of the 90.2 kDa monomer. However, the LF crystal structure PDB 1pwu (a generally V-shaped molecule with equal arm lengths ≈ 70 Å) with the same model fits the dimensions 30 Å × 48 Å × 104 Å. Given the large changes in the long and short dimensions, straightforward physical modeling of the solution structure from the crystal form is unable to match the SANS results. Full article

Beneficial bacteria are recognised for their antimicrobial compounds, making them valuable for disease control in agriculture. Bacillus species stand out for their stability, versatility, and selectivity as biocontrol agents. This study aimed to identify potential antagonists within the rhizosphere microorganisms by isolating bacterial strains from grapevine roots and rhizosphere soil in Moroccan vineyards. The antimicrobial activities of these isolates against Allorhizobium vitis, the causative agent of grapevine crown gall, were evaluated in vitro using a disc diffusion assay, followed by in planta assessments under preventive and simulated inoculation conditions. Screening led to the isolation of 123 strains, with six showing strong antagonistic properties, achieving inhibition percentages up to 39.6%. 16S rRNA sequencing led to identifying five Bacillus species: B. amyloliquefaciens, B. velezensis, B. halotolerans, B. subtilis, and B. anthracis. These strains were further characterised by their biochemical traits and plant growth-promoting abilities. Compatibility assays identified optimal combinations for microbial consortia, demonstrating pathogen inhibition up to 37.4%. In planta bioassays confirmed the effectiveness of the isolates and consortia, reducing tumour size. These findings highlight the potential of these Bacillus strains as biocontrol agents and underscore the value of microbial consortia as a sustainable approach to managing grapevine crown gall. Full article

This study tested the efficacy of xenomonitoring using contaminated flies and cockroaches at ports in Shenzhen by analysing sample data from imported flies and cockroaches from October 2023 to April 2024 to identify the pathogens they carried. Among all the samples of flies and cockroaches collected, Musca domestica vicina and Blattella germanica accounted for the highest proportion, 27.59% and 66.47%, respectively. Their positive rates for carrying Staphylococcus aureus were also the most significant, reaching 4.35% and 6.47%, respectively. The imported flies and cockroaches mainly came from Asia, with the highest proportion coming from Hong Kong, at 97.71% and 92.11%, respectively. Metagenomic sequencing indicated that the pathogens carried by the flies and cockroaches from different regions of Asia were generally similar but showed some differences. Flies from Southeast Asia, East Asia, South Asia, and West Asia and cockroaches from Southeast Asia, East Asia, and West Asia harboured unique opportunistic pathogens capable of causing gastrointestinal and respiratory infections in humans. Specifically, flies carried pathogens such as Campylobacter jejuni, Bacillus anthracis, Bacteroides fragilis, and Bordetella bronchiseptica, while cockroaches carried B. fragilis, Clostridium tetani, and Bacillus cereus. Our findings provide data support for future risk assessments of pathogens carried by imported vectors. Full article

Bacillus anthracis is a deadly pathogen that under unfavourable conditions forms highly resistant spores which enable them to survive for a long period of time. Spores of B. anthracis are transmitted through the contaminated soil or animal products and enter to the host through the skin, lungs or oral route and can cause cutaneous, injection, inhalation and gastrointestinal anthrax, respectively. The disease is caused by the toxin which is produced by them once they germinate within the host cell. Anthrax toxin is the major virulence factor which has the ability to kill the host cell. The role of protein kinases and phosphatases of B. anthracis in toxin production and other virulence related properties have also been reported. There are two vaccines, BioThrax and CYFENDUS^TM, which are approved by the FDA-USA to prevent anthrax disease. Recently, anthrax toxin has also been shown to be a potential candidate for cancer therapeutics. Through present review, we aim to provide insights into sporulation, transmission and pathogenesis of B. anthracis as well as the current state of its prevention, treatment, vaccines and possible therapeutic uses in cancer. Full article

Anthrax represents a disease resulting from infection by toxin-secreting bacteria, Bacillus anthracis. This research aimed to identify new therapeutic targets to combat anthrax. We performed assays to assess cell viability, apoptosis, glycogen consumption, and compound uptake and release in hepatocytes and cardiomyocytes responding to anthrax toxins. Microarray analysis was carried out to identify the genes potentially involved in toxin-induced toxicity. Knockdown experiments were performed to validate the contributions of the identified genes. Our study showed that anthrax edema toxin (EdTx) and lethal toxin (LeTx) induced lethal damage in mouse liver and heart, respectively. Microarray assays showed that 218 genes were potentially involved in EdTx-mediated toxicity, and 18 genes were potentially associated with LeTx-mediated toxicity. Among these genes, the knockdown of Rgs1, Hcar2, Fosl2, Hcar2, Cxcl2, and Cxcl3 protected primary hepatocytes from EdTx-induced cytotoxicity. Plasminogen activator inhibitor 1 (PAI-1)-encoding Serpine1 constituted the most significantly upregulated gene in response to LeTx treatment in mouse liver. PAI-1 knockout mouse models had a higher tolerance to LeTx compared with wild-type counterparts, suggesting that PAI-1 is essential for LeTx-induced toxicity and might represent a therapeutic target in LeTx-induced tissue damage. These results provide potential therapeutic targets for combating anthrax-toxin-induced liver and heart damage. Full article

Background: Anthrax is a serious disease caused by Bacillus anthracis (B. anthracis) with a very high mortality when the spores of B. anthracis are inhaled (inhalational anthrax). Aerosolized B. anthracis spores can be used as a deadly bioweapon. Vaccination against anthrax is the only effective preventive measure and, hence, the anthrax vaccine was administered to United States (and other) troops during the 1990–91 Gulf War. However, the anthrax vaccine is not harmless, and the anthrax vaccination has been linked to the occurrence and severity of Gulf War Illness (GWI), a debilitating Chronic Multisymptom Illness (CMI). We hypothesized that this is partly due to the combination of two factors, namely (a) the cytotoxicity of the antigen (anthrax Protective Antigen, PA) contained in the vaccine, and (b) the Human Leukocyte Antigen (HLA) genotype of susceptible vaccinees, reducing their ability to make antibodies against the cytotoxic PA. Method: Here, we tested this hypothesis by determining the association between severity of GWI symptoms in 458 GW veterans and the overall strength of the binding affinity of the PA epitopes to the specific six Human Leukocyte Antigen (HLA) Class II alleles carried by each individual (two of each of the HLA-II genes: DPB1, DQB1, DRB1), responsible for initiating the process of antibody production in otherwise immunocompetent individuals, estimated in silico. Results: We found that the severity of GWI symptomatology was negatively and significantly correlated with the strength of the predicted binding affinity of PA peptides to HLA-II molecules ($r=-0.356, p<0.001)$; the stronger the overall binding affinity, the weaker the symptoms. Since the binding of a peptide to an HLA-II molecule is the first and necessary step in initiating the production of antibodies, the findings above support our hypothesis that the severity of GWI symptomatology is partly due to a lack of HLA-II protection. Conclusions: Reduced HLA protection against the toxic anthrax vaccine may underlie GWI. Full article

This study proposes a risk analysis model based on the principles of ISO 31000 and decision theory for biological agents with potential for offensive use in Brazil. Bibliographic research was conducted on the main models already published on the subject. The German risk classification system was adopted as the main reference because it is adjustable and adopts a semiquantitative approach. After translating and adapting this model to the Brazilian context, the analytic hierarchy process (AHP) was applied to generate a hierarchical tree with criteria and subcriteria to be considered in the risk assessment. A questionnaire was administered to a group of experts to evaluate the relevant criteria and subcriteria and the risks related to three biological agents (Bacillus anthracis, Yersinia pestis, and Francisella tularensis), which were selected in an exploratory manner to exemplify the application of the model. According to the results, the criterion of impact of a biological attack was given greater importance when compared with the probability of occurrence of this event. According to the subcriteria, the greatest weight was attributed to human health when compared with agriculture, the environment, or the economy. Finally, in the evaluation of the three pathogens, B. anthracis was identified as having the highest risk for offensive use in Brazil. This research is focused on a practical approach and can be implemented by competent agencies to foster their capacity for biological defense by determining which pathogens represent the greatest risk to society. Full article

The anthrax pathogen Bacillus anthracis can remain dormant as spores in soil for many years. This applies to both natural foci and to sites of anthropogenic activity such as tanneries, abattoirs, or wool factories. The A.Br.075 (A-branch) clade (also known as A.Br.Sterne) is prominent not only because it comprises several outbreak strains but even more so because spore preparations of its namesake, the Sterne strain, are counted among the most utilized anthrax animal vaccines. In this study, we genome-sequenced and analyzed 56 additional B. anthracis isolates of the A.Br.075 clade. Four of these we recently retrieved from soil samples taken from a decades-long abandoned tannery. The other 52 strains originated from our archival collection from the 20th century. Notably, the extended phylogeny of the A.Br.075 clade indicated that many of the newly added chromosomes represent basal members, some of which are among the most basal strains from this lineage. Twelve new strains populate a very deep-branching lineage we have named A.Br.Ortho-Sterne (also known as A.Br.076). A further 11 isolates amend the clade named A.Br.Para-Sterne (A.Br.078). Finally, some of the terminal clusters of the clade named A.Br.Eu-Sterne appear to be replete with (near) identical isolates, possibly a result of widespread use of the Sterne vaccine and of its re-isolation from vaccination-related animal anthrax outbreaks. From the accrued new phylogenetic information, we designed and tested a variety of new SNP-PCR assays for rapid and facile genotyping of unassigned B. anthracis genomes. Lastly, the successful isolation of live B. anthracis from a long-abandoned tannery reemphasizes the need for continued risk awareness of such sites. Full article

Anthrax is a zoonotic disease characterized by rapid onset with usual fatal outcomes in livestock and wildlife. In Ethiopia, anthrax is a persistent disease; however, there are limited data on the isolation and molecular characterization of Bacillus anthracis strains. This study aimed to characterize B. anthracis isolated from animal anthrax outbreaks between 2019 and 2024, from different localities in Ethiopia. B. anthracis was identified using standard microbiology techniques and confirmed by real-time PCR. For the first time in Ethiopia, the genetic diversity of five Bacillus anthracis strains, isolated from dead cattle and goats, was investigated by Whole Genome Sequencing (WGS) and bioinformatics analyses. The five sequenced strains were compared to one Ethiopian B. anthracis genome and the other 29 B. anthracis genomes available in the global genetic databases to determine their phylogeny. The genomes of the strains were also analyzed to detect the presence of antimicrobial resistance and virulence genes. The whole genome SNP analysis showed that the Ethiopian B. anthracis strains were grouped in the A clade. Three strains (BA2, BA5, and BA6) belonged to the A.Br.034 subgroup (A.Br.005/006), and two strains (BA1 and BA4) belonged to the A.Br.161 (Heroin) clade of the Trans-Eurasian (TEA) group. The findings of this study will contribute to expanding the current understanding of the anthrax hotspots in Ethiopia, and the phylogenetic correlation and/or diversity of the circulating strains. Full article

The COVID-19 and mpox crisis has reminded the world of the potentially catastrophic consequences of biological agents. Aside from the natural risk, biological agents can also be weaponized or used for bioterrorism. Dissemination in a population or among livestock could be used to destabilize a nation by creating a climate of terror, by negatively impacting the economy and undermining institutions. The Centers for Disease Control and Prevention (CDC) classify biological agents into three categories (A or Tier 1, B and C) according to the risk they pose to the public and national security. Category A or Tier 1 consists of the six pathogens with the highest risk to the population (Bacillus anthracis, Yersinia pestis, Francisella tularensis, botulinum neurotoxins, smallpox and viral hemorrhagic fevers). Several medical countermeasures, such as vaccines, antibodies and chemical drugs, have been developed to prevent or cure the diseases induced by these pathogens. This review presents an overview of the primary medical countermeasures, and in particular, of the antibodies available against the six pathogens on the CDC’s Tier 1 agents list, as well as against ricin. Full article

Anthrax is a fatal zoonotic disease caused by exposure to Bacillus anthracis spores. The CDC’s guidelines divide anthrax treatment into three categories according to disease progression: post-exposure prophylaxis (PEP), systemic, and systemic with a suspicion of CNS infection. While the prognosis for PEP or the early treatment of systemic anthrax is very good, ingress of the bacteria into the CNS poses a substantial clinical challenge. Here, we use rabbits to test the efficacy of a combined treatment of meropenem and doxycycline, which is the first choice in the CDC recommendations for treating systemic patients with an indication of CNS infection. In addition, we test the efficacy of the first-generation cephalosporin, cefazolin, in treating different stages of the disease. We found that the combination of doxycycline and meropenem is highly effective in treating rabbits in our inhalation model. Cefazolin was efficient only for PEP or systemic-stage treatment and not for CNS-infected animals. Our findings support the CDC recommendation of using a combination of doxycycline and meropenem for systemic patients with or without indications of CNS infection. We found that cefazolin is a decent choice for PEP or early-stage systemic disease but recommend considering using this antibiotic only if all other options are not available. Full article

Bacillus anthracis causes anthrax through virulence factors encoded on two plasmids. However, non-B. anthracis organisms within the closely related, environmentally ubiquitous Bacillus cereus group (BCG) may cause an anthrax-like disease in humans through the partial adoption of anthrax-associated virulence genes, challenging the definition of anthrax disease. To elucidate these phenomena and their evolutionary past, we performed whole-genome sequencing on non-anthracis BCG isolates, including 93 archival (1967–2003) and 5 contemporary isolates (2019–2023). We produced annotated genomic assemblies and performed a pan-genome analysis to identify evidence of virulence gene homology and virulence gene acquisition by linear inheritance or horizontal gene transfer. At least one anthrax-associated virulence gene was annotated in ten isolates. Most homologous sequences in archival isolates showed evidence of pseudogenization and subsequent gene loss. The presence or absence of accessory genes, including anthrax-associated virulence genes, aligned with the phylogenetic structure of the BCG core genome. These findings support the hypothesis that anthrax-associated virulence genes were inherited from a common ancestor in the BCG and were retained or lost across different lineages, and contribute to a growing body of work informing public health strategies related to anthrax surveillance and identification. Full article