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Bacillus anthracis, the causative agent of anthrax, is able to exist both in environmental conditions (soil) and in the macroorganism. The manifestation of pathogenic properties of B. anthracis strains is determined by relevant plasmid composition, because the main toxin and the capsule-related virulence factors are located in bacterial plasmid. Modeling anthrax infection in vitro in macrophage culture might reveal an influence of individual B. anthracis strain characteristics on infection and development of infectious process. The aim of this study was to analyze cytokine level during infection of in vitro macrophage cell cultures with spores of anthrax microbe strains bearing varying plasmid composition. The dependence of the macrophage cell cytokine profile on the plasmid composition of B. anthracis strains was revealed while modeling anthrax infection in vitro. The presence of the toxin-producing plasmid pXO1 in anthrax microbe strains has a powerful stimulating effect on the production of macrophages J774A cell line cytokines. B. anthracis strains lacking the pXO1 plasmid virtually stimulated no production of IL-1, caused very low secretion of IL-1, IL-6, MCP-1, MIP-1, MIP-1, IL-12 (p70) and active G-CSF products. The low cytokine response of macrophage cells infected with monoplasmid strains bearing only the capsule-forming plasmid was due not only to the absence of a binary toxin, but also to disturbed regulation of capsule production associated with the absence of the atxA gene. The capsule, along with lethal and edematous toxins, belongs to the main virulence factors of B. anthracis, but strains lacking the pXO1 virulence plasmid, had its production impaired, because the main regulator of capsule synthesis is the atxA gene localized on the pXO1 plasmid being positively regulated by the acpA and acpB genes, so that strains lacking the toxin-forming plasmid, even in the presence of the encapsulation plasmid, elicit a weak cytokine response in infected cells. Diplasmid strains of B. anthracis, due to produced main virulence factors a two-component toxin and a capsule, enforce macrophages (in the experiment) to actively produce IL-1, IL-6, MCP-1, G-CSF, MIP-1; MIP-1, IL-12 (p70). Strains with moderate virulence and capable of capsulation in air virtually did not differ from highly virulent strains in terms of their effect on in vitro macrophage culture.

Anthrax is a priority zoonosis for control in Vietnam. The geographic distribution of anthrax remains to be defined, challenging our ability to target areas for control. We analyzed human anthrax cases in Vietnam to obtain anthrax incidence at the national and provincial level. Nationally, the trendline for cases remained at ≈61 cases/year throughout the 26 years of available data, indicating control efforts are not effectively reducing disease burden over time. Most anthrax cases occurred in the Northern Midlands and Mountainous regions, and the provinces of Lai Chau, Dien Bien, Lao Cai, Ha Giang, Cao Bang, and Son La experienced some of the highest incidence rates. Based on spatial Bayes smoothed maps, every region of Vietnam experienced human anthrax cases during the study period. Clarifying the distribution of anthrax in Vietnam will enable us to better identify risk areas for improved surveillance, rapid clinical care, and livestock vaccination campaigns.

The aim of the work was to characterize the epidemiological and epizootic situation on anthrax among population and animals in the Republic of Tatarstan over a period of 1920–2020.Materials and methods. The analysis of the epidemiological and epizootic situation is based on the archival data, epidemiological maps of anthrax patients, results of epizootiological-epidemiological survey of anthrax foci conducted by the Rospotrebnadzor Administration in the Republic of Tatarstan and Center of Hygiene and Epidemiology in the Republic of Tatarstan, materials of the Main Directorate of Veterinary Medicine of the Republic of Tatarstan. Microbiological studies of samples from patients and environmental objects were performed in accordance with the requirements of MR 4.2.2413-08 “Laboratory diagnostics and detection of anthrax pathogen”, real-time PCR was set using the AmpliSense Bacillus anthracis-FRT test-system (Central Research Institute of Epidemiology, Moscow). Statistical data processing was carried out using the quantile ranking method.Results and discussion. There are more than 1000 anthrax soil foci in the Republic of Tatarstan, which territorially belongs to the Volga Federal District. Analysis of the epizootic and epidemiological situation in the Republic of Tatarstan over the period of 1920–2020 has revealed that it has undergone significant changes, from mass diseases in animals and humans in early 20th century to sporadic cases of infection among population and animals at the beginning of the 21st century, primarily due to preventive veterinarysanitary measures, including veterinary and sanitary examination of animal products, mass specific immunization of animals against anthrax, arrangement of anthrax cattle burial grounds. In view of the improvement of epizootiological situation and implementation of preventive measures, there was a decrease in the incidence of anthrax among the population. The regions of the Republic have been ranked by the number of animal anthrax cases.

The relevance of the study is conditioned upon the fact that outbreaks of anthrax are periodically recorded on the territory of Ukraine, not only in ruminants, but also in pigs, fur animals, dogs, and people. The purpose of the study is to investigate the protective properties of the experimental vaccines and the abacillary vaccine “Antracol” and to prove the immunogenic effect of the extracellular toxin from the B. anthracis K-79 Z strain. Cultures of vaccine strains of anthrax were used for the experiments: B. anthracis 55, B. anthracis SB, B. anthracis K-79 Z and the “Antracol” vaccine (experimental development). Microbiological, clinical-biological, and biotechnological research methods were used in the study. The protective effect was investigated on guinea pigs (Cavia porcellus). An acute experiment was performed with a virulent strain B. anthracis 92 Z. Exotoxin was obtained from the specified cultures. The titre of the exotoxin was found in the disk precipitation reaction. The highest result regarding exotoxin production was recorded in B. anthracis K-79 Z 1 : 128 with a total protein concentration of 0.19 mg/ml, while the exotoxin of B. anthracis strain 55 with a titre of 1 : 32 showed a high total protein concentration of 0.4 mg/ml. The effect of B. anthracis exotoxins on the body was investigated by administering them to laboratory animals in different titres of exotoxins, followed by infection with the pathogenic strain B. anthracis 92 Z. The exotoxin of the vaccine strain B. anthracis K-79 Z in a titre of 1 : 64-1 : 128 shows the best protective properties against the pathogenic strain. It was found that the vaccine strains of B. anthracis SB and B. anthracis K-79 Z have the same level of protection of laboratory animals during experimental infection, which is 60%, while the vaccine from the strain B. anthracis 34F2 showed a level of protection of 20%. Based on the results of the study, it was found appropriate to use exotoxin B. anthracis in the development of prophylactic preparations against anthrax. The research results can be used by scientists and specialists in the field of veterinary medicine to develop new and improve the available vaccines for effective anthrax prevention

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.

Bacillus cereus G9241 was isolated from a welder who survived a pulmonary anthrax-like disease. Strain G9241 carries two virulence plasmids, pBCX01 and pBC210, as well as an extrachromosomal prophage, pBFH_1. pBCX01 has 99.6% sequence identity to pXO1 carried by Bacillus anthracis and encodes the tripartite anthrax toxin genes and atxA, a mammalian virulence transcriptional regulator. This work looks at how the presence of pBCX01 and temperature may affect the lifestyle of B. cereus G9241 using a transcriptomic analysis and by studying spore formation, an important part of the B. anthracis lifecycle. Here we report that pBCX01 has a stronger effect on gene transcription at the mammalian infection relevant temperature of 37°C in comparison to 25°C. At 37°C, the presence of pBCX01 appears to have a negative effect on genes involved in cell metabolism, including biosynthesis of amino acids, whilst positively affecting the transcription of many transmembrane proteins. The study of spore formation showed B. cereus G9241 sporulated rapidly in comparison to the B. cereus sensu stricto type strain ATCC 14579, particularly at 37°C. The carriage of pBCX01 did not affect this phenotype suggesting that other genetic elements were driving rapid sporulation. An unexpected finding of this study was that pBFH_1 is highly expressed at 37°C in comparison to 25°C and pBFH_1 expression leads to the production of Siphoviridae-like phage particles in the supernatant of B. cereus G9241. This study provides an insight on how the extrachromosomal genetic elements in B. cereus G9241 has an influence in bacterial phenotypes. [ABSTRACT FROM AUTHOR]

Intracellular protein–protein interactions in aberrant signaling pathways have emerged as a prime target in several diseases, particularly cancer. Since many protein–protein interactions are mediated by rather flat surfaces, they can typically not be interrupted by small molecules as they require cavities for binding. Therefore, protein drugs might be developed to compete with undesired interactions. However, proteins in general are not able to translocate from the extracellular side to the cytosolic target site by themselves, and thus an efficient protein translocation system, ideally combining efficient translocation with receptor specificity, is in high demand. Anthrax toxin, the tripartite holotoxin of Bacillus anthracis, is one of the best studied bacterial protein toxins and has proven to be a suitable candidate for cell-specific translocation of cargoes in vitro and in vivo. Our group recently developed a retargeted protective antigen (PA) variant fused to different Designed Ankyrin Repeat Proteins (DARPins) to achieve receptor specificity, and we incorporated a receptor domain to stabilize the prepore and prevent cell lysis. This strategy had been shown to deliver high amounts of cargo DARPins fused behind the N-terminal 254 amino acids of Lethal Factor (LFN). Here, we established a cytosolic binding assay, demonstrating the ability of DARPins to refold in the cytosol and bind their target after been translocated by PA. [ABSTRACT FROM AUTHOR]

2,6-Pyridine dicarboxylic acid (DPA) is considered one of the main anthrax biomarkers, and the detection of DPA is of great significance. This work presents a Zn(II)-based metal–organic framework (MOF) with the formula {[Zn2(2,6-NBC)2(vlpy)Zn(2,6-NBC)(vlpy)0.5]·0.8(2,6-H2NBC)·H2O}n (1) assembled from 2,6-naphthalenedicarboxylic acid (2,6-H2NBC) and 4,4′-vinylenedipyridine (vlpy) for a photoluminescence sensor matrix for DPA detection. 1 features a 3D pillar–layer framework with nanopore sizes of ca.13.165 Å × 12.731 Å, 12.725 Å × 11.018 Å and 13.114 Å × 13.165 Å along the three directions, occupied by lattice water and disordered 2,6-H2NBC molecules. The obtained 1 can be used as a turn-on fluorescence sensor for the detection of DPA with high selectivity, excellent sensitivity and recyclability. The luminescence of 1 demonstrates an obvious color change from blue to purplish blue as the DPA concentration rises. Furthermore, a linear correlation is presented between the fluorescence and a low DPA concentration of 0–0.3 mM, and the detection limit can reach as low as 128 nM, much lower than that of an infectious dose to a human of anthrax spores (60 μM). A fluorescence test paper is fabricated to detect DPA rapidly through color change. DFT calculations indicate the intermolecular photoinduced electron-transfer transition and hydrogen-bonding interaction between DPA molecules and the skeleton of 1 induces the "turn-on" fluorescence sensing of DPA behavior. [ABSTRACT FROM AUTHOR]

Anthrax is a deadly disease for animals and humans, which occurs mainly acutely - in a septicemic form, with inflammatory-necrotic processes in different organs. It is caused by Bacillus anthracis. The disease is widespread in Asia, Africa, Central and South America and southern Europe. In our country, until the end of the 20th century, anthrax in animals was a serious problem, but with the implementation of systematic, annual vaccinations of susceptible animals in stationary outbreaks and strict diagnostic control in the National Reference Laboratory (NRL), the incidence dropped sharply and now only single cases. The article presents the most important characteristics of the causative agent, epizootological features of the infection, pathogenesis and clinical manifestations of the disease. The diagnostic methods used and the possibilities for immunoprophylaxis and control of anthrax in animals were reviewed, in accordance with the requirements of Ordinance No. 6/October 5, 2020. Modern molecular methods for detection and identification of B. anthracis are indicated. The application of B. anthracis produced lethal factor (LF) and protective antigen (PA), as an alternative in cancer therapy is described. Anthrax is discussed in the context of the One Health concept. [ABSTRACT FROM AUTHOR]

Specific knowledge on the distribution of anthrax, a zoonosis caused by Bacillus anthracis, in Southeast Asia, including Vietnam, remains limited. In this study, we describe disease incidence and spatial distribution of human and livestock anthrax using spatially smoothed cumulative incidence from 2004 to 2020 in Cao Bang province, Vietnam. We employed the zonal statistics routine a geographic information system (GIS) using QGIS, and spatial rate smoothing using spatial Bayes smoothing in GeoDa. Results showed higher incidence of livestock anthrax compared with human anthrax. We also identified co-occurrence of anthrax in humans and livestock in northwestern districts and the province center. Livestock anthrax vaccine coverage was <6% and not equally distributed among the districts of Cao Bang province. We provide implications for future studies and recommend improving disease surveillance and response through data sharing between human and animal health sectors. [ABSTRACT FROM AUTHOR]

Anthrax is an acute infectious zoonotic disease caused by Bacillus anthracis that mostly affects grazing livestock and wildlife. Furthermore, B. anthracis is considered one of the most important biological agents of bioterrorism that could also be potentially misused in biological weapons. The distribution of anthrax in domestic animals and wildlife in Europe with a particular focus on Ukraine as a country of war was analyzed. Between 2005 and 2022, 267 anthrax cases were registered at the World Organization of Animal Health (WOAH) in animals in Europe, including 251 cases in domestic animals and 16 in wildlife. The highest numbers of cases were recorded in 2005 and 2016 followed by 2008, and the highest numbers of registered cases were reported from Albania, Russia, and Italy. In Ukraine, anthrax is currently a sporadic infection. Since 2007, 28 notifications were registered, with isolates mainly from soil samples. The highest number of confirmed anthrax cases was registered in 2018, and Odesa, which is close to Moldova, had the highest number of cases, followed by the Cherkasy region. The presence of thousands of biothermal pits and burial grounds of fallen cattle nationwide favors the re-emergence of new foci. Most confirmed cases were in cattle; however, single cases were confirmed in dogs, horses, and pigs. Further investigation of the disease in wildlife and in environmental samples is needed. The genetic analysis of isolates, investigation of susceptibility to antimicrobial compounds, and determination of virulence and pathogenicity factors are required in this volatile region of the world for awareness raising and preparedness. [ABSTRACT FROM AUTHOR]

Under certain conditions, molecular methods of diagnosis have a significant advantage over long-term classical microbiological methods. Because of the highly pathogenic properties of Bacillus anthracis, it is necessary to put into practice express methods of the pathogen identification in сase of a biological threat of the infection expand among vulnerable animals, people, and contamination of the territory. The identification of B. anthracis is difficult because of the spore and vegetative forms and the similarity to closely related species. Ukraine is a disadvantaged country for anthrax. The introduction of reliable, sensitive and specific molecular diagnostic methods is a priority in the issue of biosafety. [ABSTRACT FROM AUTHOR]

The causative microorganism in anthrax is Bacillus anthracis and this disease is more common in some regions of Türkiye. Changes in trace elements other than iron in anthrax infection have not been studied. In this study, iron, copper, lead, zi nc, manganese, magnesium, cadmium and cobalt levels were investigated in cases with cutaneous anthrax. Fifteen patients with cutaneous anthrax and 15 healthy individuals wereincluded in the study. The groups were similar to each other in terms of age and gender. Anthrax was diagnosed according to contact status with animals, symptoms, examination, and microbiological results. We performed our study with Atomic Absorption Spectrophotometer (UNICAM-929 spectrophotometer). Serum iron, lead, and cadmium levels were significantly higher in the patients than in the control subjects (p<0.05). Serum magnesium, manganese, zinc, copper, and cobalt levels were significantly lower in the patients than in the control subjects (p<0.05). The copper/zinc molar ratio was not significantly increased in the patients with cutaneous anthrax than in the control subjects. It has been reported that iron, cadmium, and lead levels are low and copper level is high in infectious diseases. But we determined the opposite situation in the patients with cutaneous anthrax. As a result, it can be said that the detection of high lead and cadmium levels in the cell in anthrax disease suppresses the immune system. Also, zinc can be used as a marker for this disease. [ABSTRACT FROM AUTHOR]

Anthrax is an important disease caused by Bacillus anthracis that threatens human and animal health worldwide. B. anthracis shows a genetically homogeneous structure because it can remain in the form of spores for a long time in nature. However, epidemiologically, it is possible to distinguish between strains with molecular methods such as multiple-locus variable-number tandem repeat analysis, single nucleotide repeat, and genome-wide single nucleotide polymorphism. In this study, it was aimed to analyze the B. anthracis MLVA-8 genotypes uploaded to the Bank for Genotyping Microbes database by multiple-locus variable-number tandem repeat analysis method. In this study, 2313 genotype profiles of 49 countries in the database were evaluated. In total, 402 genotypes were distributed within 24 clonal groups. Genotypes belonging to Turkey, on the other hand, show distribution in 8 groups. The highest number of genotypes (31%) among Turkey genotypes were localized in the XXIV clonal group. A total of 63.2% of the genotypes detected as a result of the analysis were included in the clonal groups IX, XXIII, and XXIV. As a result, among the genotypes in the MLVA Bank for Genotyping Microbes database, it was determined that the genotypes of Turkey were close to the genotypes reported from many countries. [ABSTRACT FROM AUTHOR]

【Objective】The present paper aimed to screen antagonistic bacterial strains of mango anthracnose, so as to lay a theoretical basis for biological control of mango anthracnose and provide strain resources.【Method】Colletotrichum asianum,the dominant species of mango anthrax, was used as the target, and 37 strains of Bacillus spp. obtained in previous studies were used for confrontation culture and in vitro inoculation test. The antagonistic strains with obvious control effect were selected and their antagonistic mechanism was analyzed.【Result】The results of confrontation test showed that the inhibitory rates of 10 antagonists against C. asianum were more than 78.00%,among which the inhibitory rates of M12-5,L18-7,L19-3,L17-3 and M11-6 reached 96.24%,94.94%,94.10%,91.25% and 90.00%,respectively. Eight strains of Bacillus spp. with obvious control effect were obtained by in vitro inoculation, among which strains L18-7 and L16-4 had the best control effect, with the average control efficiency of 83.00% and 79.16%,respectively. There was no significant difference between them(P＞0.05). Combined with morphological characteristics, 16S rDNA sequence and phylogenetic tree analysis based on rpoB gene sequence, strain L18-7 could be identified as B. velezensis. Strain L18-7 could produce extracellular protease and cellulase, and could produce active antagonists in five media including Luria-bertani(LB),beef peptone(NA) and potato glucose agar(PDA) etc. Strain L18-7 carried genes related to antibiotic production such as mycB,ituA,yndJ,fenB and bacA.【Conclusion】The antagonistic bacterium strain L18-7 of mango anthracnose was identified as B. velezensis. The strain grew well on LB medium at 28 ℃.It could produce antagonistic substances, extracellular protease and cellulase and carried related genes of antibiotics such as mycB,ituA,yndJ,fenB and bacA. It can be used as germplasm resources for biocontrol bacteria. [ABSTRACT FROM AUTHOR]

Anthrax is a disease that affects livestock, wildlife, and humans worldwide; however, its relative impacts on these populations remain underappreciated. Feral swine (Sus scrofa) are relatively resistant to developing anthrax, and past serosurveys have alluded to their utility as sentinels, yet empirical data to support this are lacking. Moreover, whether feral swine may assist in the dissemination of infectious spores is unknown. To address these knowledge gaps, we intranasally inoculated 15 feral swine with varying quantities of Bacillus anthracis Sterne 34F2 spores and measured the seroconversion and bacterial shedding over time. The animals also were inoculated either one or three times. The sera were evaluated by enzyme-linked immunosorbent assay (ELISA) for antibodies against B. anthracis, and nasal swabs were cultured to detect bacterial shedding from the nasal passages. We report that the feral swine developed antibody responses to B. anthracis and that the strength of the response correlated with the inoculum dose and the number of exposure events experienced. Isolation of viable bacteria from the nasal passages of the animals throughout the study period suggests that feral swine may assist in the spread of infectious spores on the landscape and have implications for the identification of environments contaminated with B. anthracis as well as the exposure risk to more susceptible hosts. [ABSTRACT FROM AUTHOR]

Aerosolized anthrax (Bacillus anthracis) spores are of extreme health concern and can remain airborne for hours and contaminate all kinds of surfaces, constituting reservoirs from which resuspension is easily produced. The assessment of decontamination techniques must therefore consider both air and surfaces. In the present study, several kinds of disinfecting fogs were experimentally tested against Bacillus thuringiensis spores, which served as a surrogate for Bacillus anthracis, both as aerosols released into the air and spread on porous and non-porous surfaces with different positions and orientations. This technology removed Bacillus thuringiensis spores from the air in 20 min with just a 1 min application of fog. The dynamics and characteristics of the fog, related to aerosol and surface interactions, proved to be critical for optimal performance and decontamination. An optimal configuration could provide effective disinfection even on indirectly reached surfaces. In all cases, 8% hydrogen peroxide (H2O2) provided a higher disinfection rate than 2% glutaraldehyde. [ABSTRACT FROM AUTHOR]

The geographical origin of a major present-day phylogenetic group (A branch WNA; A.Br.WNA) of American Bacillus anthracis is controversial. One hypothesis postulated that the anthrax pathogen reached North America via a then-existing land bridge from northeastern Asia thousands of years ago. A competing hypothesis suggested that B. anthracis was introduced to America a couple of hundred years ago, related to European colonization. The latter view is strongly supported by genomic analysis of a group of French B. anthracis isolates that are phylogenetically closely related to the North American strains of the A branch A.Br.WNA clade. In addition, three West African strains also belong to this relationship group. Recently, we have added a Spanish strain to these close relatives of the WNA lineage of American B. anthracis. Nevertheless, the diversity of Spanish B. anthracis remains largely unexplored, and phylogenetic links to European or American relatives are not well resolved. Here, we genome sequenced and characterized 29 new B. anthracis isolates (yielding 18 unique genotypes) from outbreaks in west central and central Spain in 2021. Applying comparative chromosomal analysis, we placed the chromosomes of these isolates within the established phylogeny of the A.Br.008/009 (A.Br.TEA) canonical SNP group. From this analysis, a new sub-clade, named A.Br.11/ESPc, emerged that constitutes a sister group of American A.Br.WNA. [ABSTRACT FROM AUTHOR]

Background: Anthrax is a zoonotic disease caused by the Bacillus anthracis bacteria, which is one of the top five important livestock diseases and the second top priority zoonotic disease, next to rabies, in Ethiopia, which remains a major problem for animals and public health in Ethiopia. This study was conducted to verify the existence of the outbreak, determine risk factors, and implement measures to control the anthrax outbreak in Farta woreda, South Gondar zone, Northwest Ethiopia in 2019. Methods: A community-based case-control study was conducted from March 25 to April 1, 2019. A structured questionnaire was used to collect data and for review of documents and discussion with livestock and health office staff. The collected data were analyzed by SPSS and presented in tables and graphs. Results: A total of 20 human anthrax cases with an attack rate of 2.5 per 1000 population were reported from the affected kebele. The age of the cases ranged from 1 month to 65 years (median age = 37.5 years). Of the total cases, 66.7% were male and 77.8% were 15 and older. The probability of developing anthrax among people who had unvaccinated animals was higher than in those who didn't have unvaccinated animals with an AOR = 8.113 (95% CI 1.685–39.056) and the probability of getting anthrax in relation to people's awareness of anthrax was AOR = 0.114 (95% CI 0.025–0.524). Conclusion: An anthrax outbreak occurred in Wawa Mengera Kebele of Farta woreda. The presence of unvaccinated animals in a household was found to be a risk factor for anthrax cases. Timely animal vaccination and strengthening health education on the vaccination of animals, mode of transmission, and disposal of dead animals are essential for preventing anthrax cases. [ABSTRACT FROM AUTHOR]

To meet the requirements of the Animal Rule, the efficacy of monotherapy with ANTHRASIL® (Anthrax Immune Globulin Intravenous (Human)) for inhalational anthrax was evaluated in blinded studies using rabbit and nonhuman primate models. Animals in both studies were randomized to treatment groups exposed to ~ 200 LD50Bacillus anthracis (Ames strain) spores by the aerosol route to induce inhalational anthrax. Rabbits (N = 50/group) were treated with either 15 U/kg ANTHRASIL or a volume-matching dose of IGIV after disease onset as determined by the detection of bacterial toxin in the blood. At the end of the study, survival rates were 2% (1 of 48) in the IGIV control group, and 26% (13 of 50) in the ANTHRASIL-treated group (p = 0.0009). Similarly, ANTHRASIL was effective in cynomolgus monkeys (N = 16/group) when administered therapeutically after the onset of toxemia, with 6% survival in the IGIV control and a dose-related increase in survival of 36%, 43%, and 70% with 7.5, 15 or 30 U/kg doses of ANTHRASIL, respectively. These studies formed the basis for approval of ANTHRASIL by FDA under the Animal Rule. [ABSTRACT FROM AUTHOR]

Anthrax is one of the most important zoonotic diseases which primarily infects herbivores and occasionally humans. The etiological agent is Bacillus anthracis which is a Gram-positive, aerobic, spore-forming, nonmotile, rod-shaped bacillus. The spores are resistant to environmental conditions and remain viable for a long time in contaminated soil, which is the main reservoir for wild and domestic mammals. Infections still occur in low-income countries where they cause suffering and economic hardship. Humans are infected by contact with ill or dead animals, contaminated animal products, directly exposed to the spores in the environment or spores released as a consequence of a bioterrorist event. Three classical clinical forms of the disease, cutaneous, gastrointestinal and inhalation, are seen, all of which can potentially lead to sepsis or meningitis. A new clinical form in drug users has been described recently and named "injectional anthrax" with high mortality (>33%). The symptoms of anthrax in the early stage mimics many diseases and as a consequence it is important to confirm the diagnosis using a bacterial culture or a molecular test. With regards to treatment, human isolates are generally susceptible to most antibiotics with penicillin G and amoxicillin as the first choice, and ciprofloxacin and doxycycline serving as alternatives. A combination of one or more antibiotics is suggested in systemic anthrax. Controlling anthrax in humans depends primarily on effective control of the disease in animals. Spore vaccines are used in veterinary service, and an acellular vaccine is available for humans but its use is limited. [ABSTRACT FROM AUTHOR]

Aims To assess low concentration hydrogen peroxide (LCHP) (H2O2) vapor dispersed with a home humidifier for its ability to decontaminate vehicle interiors contaminated with Bacillus anthracis surrogate Bacillus atrophaeus spores. Methods and results Efficacy of a vaporized 3% H2O2 solution was evaluated for liquid volumes, on/off vehicle heating, ventilation, and air conditioning (HVAC) system operations, and temperatures that ranged from 5 to 27°C. Survival of the spores was assessed by quantification of remaining viable spores with efficacy quantified in terms of mean log10 reduction. Decontamination efficacy after the 6-day dwell time increased when the 3% H2O2 liquid volume was doubled, increasing from 4-of-10 to 10-of-10 nondetects (zero colonies counted using standard dilution and filter plating) inside the vehicle cabin. Recirculating cabin air through the HVAC system during decontamination decreased efficacy to 6-of-10 non-detects. While no 6-log10 reduction in viable spores was observed on the cabin filter with the cabin filter kept in place, a 6-log10 reduction was achieved after its removal and placement in the cabin during treatment. Conclusions Results from this study allow for informed decisions on the use of LCHP vapor as an effective decontamination approach for vehicle interiors. [ABSTRACT FROM AUTHOR]

Objectives To evaluate the in vitro activity and in vivo efficacy of delafloxacin against Bacillus anthracis , the causative agent of anthrax. Methods MICs were obtained according to CLSI guidelines for 30 virulent isolates and 14 attenuated antibiotic-resistant strains. For the in vivo efficacy study, mice were administered delafloxacin (30–62.5 mg/kg) subcutaneously, or ciprofloxacin (30 mg/kg) intraperitoneally beginning at either 24 or 48 ± 1 h post-challenge (post-exposure prophylaxis) and continued every 12 h for 14 days with study termination on day 30. The mean inhaled dose in the study was approximately 103 × LD50 equivalents, and the range was 87–120 × LD50. Results Delafloxacin (MIC90 = 0.004 mg/L) was 16-fold more potent than ciprofloxacin (MIC90 = 0.06 mg/L) against a 30-strain set of virulent B. anthracis. Against a panel of attenuated antibiotic-resistant strains, delafloxacin demonstrated potency ≥128-fold over that observed with ciprofloxacin. When evaluated in vivo , mice treated with all delafloxacin doses tested at 24 h post-challenge demonstrated equivalent survival compared with mice treated with the positive control ciprofloxacin. Because of the high challenge dose of spores, mice treated at 48 h showed rapid and high mortality in all groups including the positive control. Surviving animals in all delafloxacin- and ciprofloxacin-treated groups (24 and 48 h) showed complete splenic clearance of infection and <2.2 × 103 cfu/g lung tissue. Conclusions Given the high bar set by the 100 × LD50 challenge dose in this study, the results from delafloxacin treatment are promising for the treatment of inhaled anthrax. [ABSTRACT FROM AUTHOR]

Bacillus anthracis, the causative agent of anthrax in humans, livestock, and wildlife, exists in a community with hundreds of other species of bacteria in the environment. Work on the genetics of these communities has shown that B. anthracis shares a high percentage of chromosomal genes with both B. thuringiensis and B. cereus, and that phenotypic differences among these bacteria can result from extra-chromosomal DNA in the form of plasmids. We developed a simple hypothetical individual-based model to simulate the likelihood of detecting plasmids with genes encoding anthrax toxins within bacterial communities composed of B. anthracis, B. thuringiensis, and B. cereus, and the surrounding matrix of extra-cellular polymeric substances. Simulation results suggest the horizontal transfer of plasmids with genes encoding anthrax toxins among Bacillus species persisting outside the host could function as a proximate factor triggering anthrax outbreaks. [ABSTRACT FROM AUTHOR]

BACKGROUND: In recent years, inductively coupled plasma mass spectrometry (ICP-MS) has been widely used in the field of molecular biology because of its unique advantages. Anthrax is a widespread and long-standing infectious disease, which affects and restricts people's work and life seriously. OBJECTIVE: The study goal is to develop a new method for the detection of anthrax. METHODS: A rapid, sensitive and accurate method for the detection of anthrax characteristic DNA was proposed by combing gold nanoparticles (AuNPs) and inductively coupled plasma mass spectrometry. RESULTS: The linear range of this method is 100–2500 pmol/L and the limit of detection of 16.61 pmol/L. CONCLUSION: The proposed method has numerous advantages, including simplicity of operation, high sensitivity, and specificity, which provides a new idea for the detection of anthrax. Importantly, this methodology has good potential for the detection of other biological substances such as bacteria and viruses by changing the modification sequence on the nanoparticle probe. [ABSTRACT FROM AUTHOR]

Bacillus cereus G9241 was isolated from a Louisiana welder suffering from an anthrax-like infection. The organism carries two transcriptional regulators that have previously been proposed to be incompatible with each other in Bacillus anthracis: the pleiotropic transcriptional regulator PlcR found in most members of the Bacillus cereus group but truncated in all B. anthracis isolates, and the anthrax toxin regulator AtxA found in all B. anthracis strains and a few B. cereus sensu stricto strains. Here we report cytotoxic and hemolytic activity of cell free B. cereus G9241 culture supernatants cultured at 25°C to various eukaryotic cells. However, this is not observed at the mammalian infection relevant temperature 37°C, behaving much like the supernatants generated by B. anthracis. Using a combination of genetic and proteomic approaches to understand this unique phenotype, we identified several PlcR-regulated toxins to be secreted highly at 25°C compared to 37°C. Furthermore, results suggest that differential expression of the protease involved in processing the PlcR quorum sensing activator molecule PapR appears to be the limiting step for the production of PlcR-regulated toxins at 37°C, giving rise to the temperature-dependent hemolytic and cytotoxic activity of the culture supernatants. This study provides an insight on how B. cereus G9241 is able to "switch" between B. cereus and B. anthracis-like phenotypes in a temperature-dependent manner, potentially accommodating the activities of both PlcR and AtxA. [ABSTRACT FROM AUTHOR]

The disease anthrax occurs generally in herbivores and the causative organism (Bacillus anthracis) infects humans who come in contact with infected animals or their products. The persistence of anthrax spores for decades and its lethality contribute to its biowarfare potential. We conducted this systematic review along with risk mapping to investigate the spatio-temporal distribution, clinico-epidemiological, socio-behavioural and programmatic issues pertaining to anthrax in India over the last two decades.Peer reviewed quantitative and qualitative studies and grey literature comprising weekly reports of the ‘Integrated Disease Surveillance Program’ (IDSP), were accessed for extracting data. IDSP data were used for geo-referencing of the villages of anthrax cases; Pseudo-absence was generated to fit a Bayesian Additive Regression Trees (BART) model to develop anthrax risk map.The case fatality rate of cutaneous anthrax ranged from 2% to 38%, while the gastrointestinal and inhalational types were 100% fatal. Our synthesis revealed that human anthrax outbreaks in India were clustered around the eastern coastal regions. The states of Odisha, West Bengal, Andhra Pradesh and Jharkhand reported maximum number of outbreaks. Odisha reported a maximum number of 439 human anthrax cases since 2009, of which Koraput district contributed to 200 cases (46%). While handling or consumption of infected animal product were proximal drivers of these events, poverty, lack of awareness, traditional beliefs and local practices served as facilitatory factors. Other structural determinants were wild life-livestock interface, historical forest loss, soil pH, soil-water balance, organic carbon content, temperature, rainfall and humidity.The programmatic issues identified through this review were lack of active surveillance, non-availability of diagnostic facility at the periphery, delayed reporting, absence of routine livestock vaccination and lack of adequate veterinary services. Interventions based on One-health approach in the country merit immediate policy and program attention; high risk zones for anthrax identified during present investigation, should be prioritized.

Vaccination against Bacillus anthracis is the best preventive measure against the development of deadly anthrax disease in the event of exposure to anthrax either as a bioweapon or in its naturally occurring form. Anthrax vaccines, however, have historically been plagued with controversy, particularly related to their safety. Fortunately, recent improvements in anthrax vaccines have been shown to confer protection with reduced short-term safety concerns, although questions about long-term safety remain. Here, we (a) review recent and ongoing advances in anthrax vaccine development, (b) emphasize the need for thorough characterization of current (and future) vaccines, (c) bring to focus the importance of host immunogenetics as the ultimate determinant of successful antibody production and protection, and (d) discuss the need for the systematic, active, and targeted monitoring of vaccine recipients for possible Chronic Multisymptom Illness (CMI).

Institut Pasteur and Bacillus anthracis have enjoyed a relationship lasting almost 120 years, starting from its foundation and the pioneering work of Louis Pasteur in the nascent fields of microbiology and vaccination, and blooming after 1986 following the molecular biology/genetic revolution. This contribution will give a historical overview of these two research eras, taking advantage of the archives conserved at Institut Pasteur. The first era mainly focused on the production, characterisation, surveillance and improvement of veterinary anthrax vaccines; the concepts and technologies with which to reach a deep understanding of this research field were not yet available. The second period saw a new era of B. anthracis research at Institut Pasteur, with the anthrax laboratory developing a multi-disciplinary approach, ranging from structural analysis, biochemistry, genetic expression, and regulation to bacterial-host cell interactions, in vivo pathogenicity, and therapy development; this led to the comprehensive unravelling of many facets of this toxi-infection. B. anthracis may exemplify some general points on how science is performed in a given society at a given time and how a scientific research domain evolves. A striking illustration can be seen in the additive layers of regulations that were implemented from the beginning of the 21st century and their impact on B. anthracis research. B. anthracis and anthrax are complex systems that raise many valuable questions regarding basic research. One may hope that B. anthracis research will be re-initiated under favourable circumstances later at Institut Pasteur.

Due to several factors, such as environmental and climate changes, the risk of health threats originating at the human–animal–environment interface, including vector-borne diseases (VBDs) and zoonoses, is increasing. Low-resource settings struggle to counter these multidimensional risks due to their already-strained health systems and are therefore disproportionally affected by the impact caused by these changes. Systemic approaches like One Health (OH) are sought to strengthen prevention and preparedness strategies by addressing the drivers of potential threats with a multidisciplinary and multisectoral approach, considering the whole system at the human–animal–environment interface. The integration of OH in national plans can be challenging due to the lack of effective coordination and collaboration among different sectors. To support the process of knowledge coproduction about the level of OH integration in prevention and preparedness strategies against health threats in Armenia, a situation analysis was performed on Crimean–Congo hemorrhagic fever/virus and anthrax (identified by local stakeholders as priorities to be addressed with the OH approach), and actions to strengthen the national OH system were identified with the support of a OH conceptual framework. The study highlighted that multidisciplinary and multisectoral efforts towards prevention and preparedness against VBDs and zoonoses threats need to be strengthened in Armenia, and priority actions to integrate the OH approach were identified.

Bacillus anthracis is a spore-forming, Gram-positive bacterium responsible for anthrax, an acute infection that most significantly affects grazing livestock and wild ungulates, but also poses a threat to human health. The geographic extent of B. anthracis is poorly understood, despite multi-decade research on anthrax epizootic and epidemic dynamics; many countries have limited or inadequate surveillance systems, even within known endemic regions. Here, we compile a global occurrence dataset of human, livestock and wildlife anthrax outbreaks. With these records, we use boosted regression trees to produce a map of the global distribution of B. anthracis as a proxy for anthrax risk. We estimate that 1.83 billion people (95% credible interval (CI): 0.59-4.16 billion) live within regions of anthrax risk, but most of that population faces little occupational exposure. More informatively, a global total of 63.8 million poor livestock keepers (95% CI: 17.5-168.6 million) and 1.1 billion livestock (95% CI: 0.4-2.3 billion) live within vulnerable regions. Human and livestock vulnerability are both concentrated in rural rainfed systems throughout arid and temperate land across Eurasia, Africa and North America. We conclude by mapping where anthrax risk could disrupt sensitive conservation efforts for wild ungulates that coincide with anthrax-prone landscapes.

Bacterial endospores produced by Bacillus and Clostridium species can remain dormant and highly resistant to environmental insults for long periods, but they can also rapidly germinate in response to a nutrient-rich environment. Multiple proteins involved in sensing and responding to nutrient germinants, initiating solute and water transport, and accomplishing spore wall degradation are associated with the membrane surrounding the spore core. In order to more fully catalog proteins that may be involved in spore germination, as well as to identify protein changes taking place during germination, unbiased proteomic analyses of membrane preparations isolated from dormant and germinated spores of Bacillus anthracis and Bacillus subtilis were undertaken. Membrane-associated proteins were fractionated by SDS-PAGE, gel slices were trypsin digested, and extracted peptides were fractionated by liquid chromatography and analyzed by matrix-assisted laser desorption ionization-tandem time of flight mass spectrometry. More than 500 proteins were identified from each preparation. Bioinformatic methods were used to characterize proteins with regard to membrane association, cellular function, and conservation across species. Numerous proteins not previously known to be spore associated, 6 in B. subtilis and 68 in B. anthracis, were identified. Relative quantitation based on spectral counting indicated that the majority of spore membrane proteins decrease in abundance during the first 20 min of germination. The spore membranes contained several proteins thought to be involved in the transport of metal ions, a process that plays a major role in spore formation and germination. Analyses of mutant strains lacking these transport proteins implicated YloB in the accumulation of calcium within the developing forespore.IMPORTANCE Bacterial endospores can remain dormant and highly resistant to environmental insults for long periods but can also rapidly germinate in response to a nutrient-rich environment. The persistence and subsequent germination of spores contribute to their colonization of new environments and to the spread of certain diseases. Proteins of Bacillus subtilis and Bacillus anthracis were identified that are associated with the spore membrane, a position that can allow them to contribute to germination. A set of identified proteins that are predicted to carry out ion transport were examined for their contributions to spore formation, stability, and germination. Greater knowledge of spore formation and germination can contribute to the development of better decontamination strategies.

Abstract Background Anthrax is a zoonotic infection caused by the bacteria Bacillus anthracis. Humans acquire cutaneous infection through contact with infected animals or animal products. On May 6, 2018, three cows suddenly died on a farm in Kiruhura District. Shortly afterwards, a sub-county chief in Kiruhura District received reports of humans with suspected cutaneous anthrax in the same district. The patients had reportedly participated in the butchery and consumption of meat from the dead cows. We investigated to determine the magnitude of the outbreak, identify exposures associated with illness, and suggest evidence-based control measures. Methods We conducted a retrospective cohort study among persons whose households received any of the cow meat. We defined a suspected human cutaneous anthrax case as new skin lesions (e.g., papule, vesicle, or eschar) in a resident of Kiruhura District from 1 to 26 May 2018. A confirmed case was a suspected case with a lesion testing positive for B. anthracis by polymerase chain reaction (PCR). We identified cases through medical record review at Engari Health Centre and active case finding in the community. Results Of the 95 persons in the cohort, 22 were case-patients (2 confirmed and 20 suspected, 0 fatal cases) and 73 were non-case household members. The epidemic curve indicated multiple point-source exposures starting on May 6, when the dead cows were butchered. Among households receiving cow meat, participating in slaughtering (RR = 5.3, 95% CI 3.2–8.3), skinning (RR = 4.7, 95% CI = 3.1–7.0), cleaning waste (RR = 4.5, 95% CI = 3.1–6.6), and carrying meat (RR = 3.9, 95% CI = 2.2–7.1) increased the risk of infection. Conclusions This cutaneous anthrax outbreak was caused by handling infected animal carcasses. We suggested to the Ministry of Agriculture, Animal Industry and Fisheries to strengthen surveillance for possible veterinary anthrax and ensure that communities do not consume carcasses of livestock that died suddenly. We also suggested that the Ministry of Health equip health facilities with first-line antibiotics for community members during outbreaks.

Abstract Background Anthrax is a zoonotic disease that causes frequent outbreaks in livestock and fatal human cases in Burkina Faso. Effective surveillance of this disease calls for the establishment of an integrated surveillance system, in line with the One Health concept. However, despite a strong technical and financial support from international partners, surveillance is still poorly conducted within an integrated approach. Based on stakeholder perspectives, the study has for objective to deepen our understanding of the anthrax surveillance system and to identify the obstacles and levers towards a more integrated approach to anthrax surveillance in Burkina Faso. Methods The data was collected from a literature review and interviews with surveillance stakeholders. We first conducted a qualitative descriptive analysis of the data to characterize the surveillance system (programmes, actors, collaboration). In a second step, we conducted a thematic analysis of the informants' discourse in order to identify what represents an obstacle or, conversely, a lever for a more integrated approach to anthrax surveillance. Results The surveillance system of anthrax in Burkina Faso includes three programmes (in the livestock, wildlife and human sectors), which involves 30 actors. These sectoral programmes operate almost independently from one another, although some collaborations are existing for the governance and implementation of surveillance activities. Analysis of the discourse of key stakeholders led to the identification of four categories of factors that may influence the implementation of an integrated surveillance system in the country: knowledge; technical, organizational and social capacities; motivation; intersectoral governance. Conclusions This study highlights the difficulty of translating One Health governance to the national level and the need to better articulate the visions of all categories of stakeholders. This study also reveals the need to develop specific evaluation systems for integrated policies in order to provide credible evidence of their added value for a better management of zoonotic diseases. Finally, our study underlines the need to act upstream the emergence of zoonoses and allocate more resources to the prevention of zoonoses than to their control.

Anthrax poses a pressing issue for veterinary medicine and public health in many countries, including the Russian Federation, which necessitates the improvement and development of new, sensitive and specific diagnostic tools.The aim of the work was to create an experimental peroxidase conjugate for the detection of specific antibodies to the anthrax pathogen and to optimize the conditions for performing enzyme immunoassay (ELISA).Materials and methods. The peroxidase conjugate was constructed using horseradish peroxidase and Staphylococcus aureus protein A (Sigma-Aldrich, USA). Bacterial antigens isolated from strains of Bacillus anthracis 55ΔTPA-1Spo, B. anthracis Sterne 34 F2 were used as sensitizing agents. The developed experimental batches of the conjugate were tested in ELISA for the ability to bind antibodies in the blood sera of anthrax patients and vaccinated individuals. The sensitivity, specificity, and accuracy of the method were calculated using the built-in functions of the ROCR software package.Results and discussion. The peroxidase conjugate to detect specific antibodies to the anthrax pathogen in the study of clinical material has been developed; conditions for the ELISA performance have been optimized. To interpret the results of the study, a threshold value of the positivity coefficient was used, below which the result was considered negative, and at an equal or higher value, positive. The test demonstrated significant differences in the “positivity coefficient” indicator for the “Healthy”/“Sick” and “Healthy”/“Vaccinated” groups, while the differences between the “Sick”/“Vaccinated” groups were statistically insignificant. The maximum accuracy of the method was observed at blood serum dilutions of 1:250 and 1:500. 100 % intra-run, run-to-run and series-to-series reproducibility has been established for all positive samples. The sensitivity and specificity of the experimental peroxidase conjugates were 100 and 95.8 %, respectively, and the accuracy was 97.6 %.