Adriana Marcos Vivoni

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.

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.

The aerobic endospore-forming bacteria (AEFB) comprise species of Bacillus and related genera, allocated in the phylum Firmicutes. Although Bacillus spp. are among the first bacteria to be characterised, the wide diversity renders appropriate categorisation and generalisations challenging tasks. To determine genetic diversity, analyses at the molecular level are the most accurate. However, gene expression, morphological, biochemical, and physiological aspects must also be considered. The metabolism of bacteria is adapted to their natural environment or host. Thus, metabolic outlines can be used for identifying AEFB, form the basis of the formal description of bacterial taxa, and are strongly recommended for taxonomic purposes. This work addressed the biochemical and physiological profiles of 312 environmental AEFB—designated as SDF (Solo do Distrito Federal)—by performing 30 tests. Out of it, 246 were classified by 16S rRNA gene sequences. We summarised the phenotypic test relationships among selected SDF strains using a Pearson correlation-based clustering represented in heatmaps. In practice, biochemical and physiological profiles are often less discriminatory than molecular data and may be unstable because of the loss of traits. Though these test reactions are not universally positive or negative within species, they may define biotypes and be efficient strain markers, enhancing the accuracy of unknown sample identification. It can be also helpful in selecting the best represent the phenotypes of samples. Along with the other phenotypic and genotypic data, the present results will be of great importance for the robust classification of the SDF strains within the scope of the polyphasic approach.

O Programa Antártico Brasileiro se desenvolve com o apoio da Marinha, através dos seus navios e da Estação Antártica Comandante Ferraz (EACF). Entretanto, ...

B. amyloliquefaciens is a Gram-positive, aerobic, motile rod, often found in soil, which has been described as a plant growth promoter and is used in several industrial processes. This study reports an episode involving the gassy spoilage of salad dressing caused by B. amyloliquefaciens in a production facility located in Rio de Janeiro, Brazil. Nine B. amyloliquefaciens strains were isolated from spoiled salad dressings, the sugar used as a raw material in the manufacture and from the production plant. A genotypic analysis of the isolates by Rep-PCR generated eight band profiles grouped in five Rep-PCR clusters. When re-inoculated into fresh salad dressing three B. amyloliquefaciens isolates belonging to the Rep-PCR clusters A, D and E were able to reproduce the gassy spoilage process, whereas the isolates belonging to the Rep-PCR clusters B and C did not produce any visible spoilage, suggesting that these isolates were not directly involved in the spoilage process. The predominant Rep-PCR cluster, cluster A, included strains isolated from barbecue and passion fruit seed salad dressings and from sugar (raw material), suggesting it is a common source of contamination for such salad dressings.

The presence of Bacillus cereus in milk is a major concern in the dairy industry. In this study 27 Bacillus cereus sensu lato isolates from pasteurized and ultrahigh-temperature (UHT) milk (24 whole UHT and 4 pasteurized samples) collected at supermarket chains in Rio de Janeiro, Brazil, were evaluated to assess the potential risk for food poisoning. Toxigenic and virulence profiles were defined by gene-specific PCR. Affiliation to phylogenetic groups was assigned by panC sequencing. Microbiological analysis revealed the presence of B. cereus s.l. in eight (33.3%) brands (six brands of UHT and two brands of pasteurized milk). Twenty-seven isolates were recovered (13 B. cereus and 14 Bacillus thuringiensis). Predominant toxigenic patterns were type I (contains all toxin genes except ces) and type II (does not contain cytK and ces), with seven (25.9%) isolates each. Predominant virulence patterns were type 2 (does not contain hlyII or shp) and type 3 (contains all virulence genes), with five (18.5%...

Bacillus thuringiensis is used to produce biopesticides against target-insects of importance in agroindustry and human health. However, B. thuringiensis can carry the same enterotoxin-encoding genes as Bacillus cereus, suggesting a potential risk of food poisoning. The objective of this work was to study the application of gamma radiation to eliminate spores in biopesticides based on B. thuringiensis serovar israelensis in order to avoid the liberation of these spores in the environment. The application of 20 kGy of radiation ensures total inactivation of the spores in the insecticide without changing the larvicidal activity. Results showed that it is possible to develop formulations containing B. thuringiensis serovar israelensis (IPS-82) eliminating viable spores with gamma radiation without affecting the larvicidal activity. Irradiated formulations would be safer for the environment and humans since the elimination of viable spores abolishes the pathogenic potential of B. thuringiensis strains. Keywords: biopesticides, gamma radiation, larvicidal insecticides.

Thirty samples of roasted ground coffee beans from 10 different commercial brands were analyzed to investigate the occurrence and levels of Bacillus cereus and Bacillus thuringiensis strains. Strains were evaluated for their genetic diversity by repetitive element sequence polymorphism PCR (Rep-PCR) and for their toxigenic profiles, i.e., the presence of hblA, hblC, hblD, nheA, nheB, nheC, cytK, ces, and entFM. Survival and multiplication of B. cereus sensu lato in the ready-to-drink coffee was determined to evaluate this beverage as a possible vehicle for B. cereus infection. B. cereus was detected in 17 (56.7%) of the 30 samples, and B. thuringiensis was detected in 8 (26.7%) of the 30 samples. Five samples did not produce any characteristic growth. The most common gene, entFM, was detected in 23 strains (92%). The NHE complex (nheA, nheB, and nheC genes) was found in 19 strains (76%). The HBL complex (hblA, hblC, and hblD) was found in 16 strains (64%). All strains were negative for ces. The cytK gene was found in 16 strains (64%). The computer-assisted cluster analysis of Rep-PCR profiles using a clustering criterion of 80% similarity revealed four main clusters. Cluster 1 was the predominant and comprised three B. thuringiensis strains with 100% similarity, cluster 2 comprised two B. cereus strains (100% similarity), cluster 3 comprised two B. thuringiensis strains (90% similarity), and cluster 4 comprised one B. thuringiensis strain and one B. cereus strain (85% similarity). The cluster analysis of fingerprints generated by Rep-PCR revealed a high genetic diversity among the B. cereus strains, suggesting that the contamination could have originated from different sources. In our experiments, when sugar was added and the beverage was kept in thermic bottles there was a significant increase in B. cereus sensu lato levels, which may increase the risk of food poisoning. These results highlight the need for additional studies on this subject to better evaluate coffee as a food poisoning vehicle.