Bacillus cereus CGMCC 1.60196: a promising bacterial inoculant isolated from biological soil crusts for maize growth enhancement.

Soil microbial inoculants are widely recognized as an environmentally friendly strategy for promoting crop growth and increasing productivity. However, research on utilizing the microbial resources from desert biological soil crusts to enhance crop growth remains relatively unexplored. In the present work, a bacterial strain designated AC1-8 with high levels of amylase, protease, and cellulase activity was isolated from cyanobacterial crusts of the Tengger Desert and identified as Bacillus cereus (CGMCC 1.60196). The refinement of the fermentation parameters of B. cereus CGMCC 1.60196 determined that the most effective medium for biomass production was composed of 5 g/L glucose, 22 g/L yeast extract and 15 g/L MgSO4, and the optimal culture conditions were pH 6.0, temperature 37°C, inoculation quantity 3% and agitation speed 240 rpm. Furthermore, the utilization of B. cereus CGMCC 1.60196 has resulted in substantial improvements in various growth parameters of maize seedlings, including shoot length, shoot fresh and dry weights, root fresh and dry weights, and the contents of chlorophyll a, chlorophyll b, and total chlorophyll. The most pronounced growth promotion was observed at an application concentration of 1 × 109 CFU/m2. These results suggest that the novel B. cereus strain, isolated from cyanobacterial crusts, can be regarded as an exemplary biological agent for soil improvement, capable of enhancing soil conditions, promoting crop cultivation and supporting food production.

Cooling Uncovered Foods at a Depth of ∼5.1 cm (2 in.) or Less Poses Little Risk of Pathogen Growth.

The U.S. Food and Drug Administration has guidelines for cooling cooked foods in retail operations. Data on foodborne illness risk factors in restaurants indicate that cooling is often out of compliance with these guidelines. We sought to identify factors under the control of the operator that had a significant effect on the cooling rates of cooked foods. Minneapolis Minnesota Health Inspectors were trained in standardized operating procedures for cooling data collection. Data loggers set to appropriate time intervals and calibrated to ±0.5 °C (∼1°F) were used in data collection. Analysis was performed using the R statistical computing language version 4.2.2. Preexisting pathogen models were used to predict Log CFU increases of Clostridium perfringens or Bacillus cereus. Data from 224 recipes were recorded by inspectors between October 2018 and October 2019. Food depth had a highly significant effect (p = 8.90E-08) on cooling rate. The use of an ice bath or ice wand was also significant (p < 0.005). There was a significant correlation between container material (metal or plastic) and food depth because foods with a greater depth are often being cooled in plastic containers. Foods at a depth greater than 5.1 cm (2 in.) that cooled faster than 0.23 log(°C)/h were often wholly or partially cooled in blast chillers or freezers, cooled using an ice bath or ice wand (or both), or were composed of protein pieces (e.g., chicken wings) that facilitated more rapid cooling due to air gaps in the food. Foods in shallow containers at a food depth of less than or equal to ∼5.1 cm (2 in.) that cooled more slowly than 0.23 log(°C)/h were being cooled at temperatures greater than 5 °C (41°F) or were partially or wholly covered. These foods also showed little evidence of pathogen growth from predictive models. Our analysis shows that cooling foods in shallow containers at a food depth of less than or equal to ∼5.1 cm (2 in.) poses little risk of significant pathogen growth.

Bacillus cereus infection of the penis: an unusual infection.

Background: Bacillus cereus (B. cereus) is a Gram-positive, rod-shaped, motile organism, found in the environment and may exist in contaminated food sources such as reheated rice, vegetables and may lead to gastrointestinal symptoms after ingestion including diarrhea, nausea, and vomiting due to enterotoxigenic and emetic toxins. Non-gastrointestinal infections of Bacillus cereus have been reported in the literature, which include cutaneous and non-cutaneous infections in immunocompetent and immunocompromised individuals. Case presentation: A 38-year-old man presented with a one-week history of penile swelling and redness that started after an episode of severe diarrhea and vomiting, which soiled his genital region few hours after vigorous intercourse with his wife. This has led to infection of the penile skin by an unusual organism: Bacillus (B.) cereus. The patient was treated using fucidic acid ointment applications for 2 weeks achieving complete recovery. Clinical discussion: The recovery of B. cereus from the penile infection in our patient reveals the first case of such an unusual infection, though this pathogen has been reported to cause a wide range of non-GI tract infections, which include bacteremia, meningitis, endocarditis, endophthalmitis, pneumonia, and soft tissue diseases. Virulence factors allow this organism to induce diarrhea in addition to having dermonecrotic, cytotoxic, hemolytic properties resulting in a wide range of dermatologic presentations. Conclusion: The authors report a unique case of penile skin infection caused by B. cereus, an unusual culprit for an uncommon presentation successfully treated with fucidic acid ointment. This is the first case in literature describing such an entity.

Effects of Na+ adaptation on Bacillus cereus endospores inactivation and transcriptome changes.

As Bacillus cereus endospores exist in various vegetables grown in soil, the possibility of contamination in food products with high salt concentrations cannot be ignored. Recent studies revealed that harsh conditions affect the resistance of bacteria; thus, we investigated the developmental aspect of heat resistance of B. cereus after sporulation with high NaCl concentration. RNA sequencing was conducted for transcriptomic changes when B. cereus endospores formed at high salinity, and membrane fluidity and hydrophobicity were measured to verify the transcriptomic analysis. Our data showed that increasing NaCl concentration in sporulation media led to a decrease in heat resistance. Also, endospore hydrophobicity, membrane fluidity, and endospore density decreased with sporulation at higher NaCl concentrations. When the transcript changes of B. cereus sporulated at NaCl concentrations of 0.5 and 7% were analyzed by transcriptome analysis, it was confirmed that the NaCl 7% endospores had significantly lower expression levels (FDR<0.05) of genes related to sporulation stages 3 and 4, which led to a decrease in expression of spore-related genes such as coat proteins and small acid-soluble proteins. Our findings indicated that high NaCl concentrations inhibited sporulation stages 3 and 4, thereby preventing proper cell maturation in the forespores and adequate formation of the coat protein and cortex. This inhibition led to decreased endospore density and hydrophobicity, ultimately resulting in reduced heat resistance.resistanceWe expect that this study will be utilized as a baseline for further studies and enhance sterilization strategies.

The exploration of pasteurization processes and mechanisms of inactivation of Bacillus cereus ATCC 14579 using radio frequency energy.

Radio frequency (RF) heating has been utilized to investigate sterilization techniques, but the mechanism of sterilization via RF heating, particularly on Bacillus cereus (B. cereus), has not been thoroughly examined. In this paper, sterilization processes and potential bactericidal mechanisms of B. cereus using RF were investigated. The best heating and sterilization efficiency was achieved at (Electrode gap 130 mm, conductivity of bacterial suspension 0.1 S/m, volume of bacterial suspension 40 mL). Heating a suspension of B. cereus to 90 °C in 80 s using RF reduced the number of viable bacteria by 4.87 logarithms. At the cellular level, there was a significant leakage of nucleic acids and proteins from the bacterial cells. Additionally, the integrity of the cell membrane was severely damaged, with a decrease in ATP concentration of 2.08 mM, Na, K-ATPase activity to 10.7 (U/109 cells), and Ca, Mg-ATPase activity to 11.6 (U/109 cells). At the molecular level, transcriptomics analysis showed that RF heating of B. cereus to 65 °C produced 650 more differentially expressed genes (DEGs) compared with RF heating to 45 °C. The GO annotation analysis indicated that the majority of differentially expressed genes (DEGs) were predominantly associated with cellular components. KEGG metabolic analysis showed enrichment in microbial metabolism in diverse environments, etc. This study investigated the potential bactericidal mechanism of B. cereus using RF, and provided some theoretical basis for the research of the sterilization of B. cereus.

Microbiological quality of vegan alternatives to dairy and meat products in England during 2022-2023.

AIMS: Plant-based alternatives to meat and dairy products have become increasingly popular in the UK. Despite a public perception that they have a relatively low microbiological risk, outbreaks of illness have been linked with these foods. This study aimed to assess the microbiological safety and quality of vegan alternatives to dairy and meat products available in England. METHODS AND RESULTS: Samples were collected between September 2022 and March 2023 from retail, production and catering premises, and tested for a range of bacterial pathogens and hygiene indicators using standard procedures. A total of 937 samples were tested, of which 92% were of a satisfactory microbiological quality, 3% were borderline and 5% were unsatisfactory. Those interpreted as unsatisfactory were due to elevated counts of Enterobacteriaceae and E. coli (indicators of poor hygiene) rather than pathogenic micro-organisms. Listeria monocytogenes was present in five samples of tofu, all from the same producer (all at counts of < 100 cfu g-1), while other Listeria species were detected at counts of < 20 cfu g-1 in two burgers and two 'vegan chicken' products. The majority of samples did not have pH and water activity values that would significantly contribute to preventing microbial growth: 62.4% had pH > 5.0 and 82.4% had Aw > 0.94. CONCLUSIONS: The majority of vegan products examined were of a satisfactory quality, but results demonstrate that microbiological control must be maintained using appropriate processing and storage temperatures and application of a safe length of shelf-life.

Organization, conservation, and diversity of biosynthetic gene clusters in Bacillus sp. BH32 and its closest relatives in the Bacillus cereus group.

This study explores the organization, conservation, and diversity of biosynthetic gene clusters (BGCs) among Bacillus sp. strain BH32, a plant-beneficial bacterial endophyte, and its closest nontype Bacillus cereus group strains. BGC profiles were predicted for each of the 17 selected strains using antiSMASH, resulting in the detection of a total of 198 BGCs. We quantitatively compared the BGCs and analysed their conservation, distribution, and evolutionary relationships. The study identified both conserved and singleton BGCs across the studied Bacillus strains, with minimal variation, and discovered two major BGC synteny blocks composed of homologous BGCs conserved within the B. cereus group. The identified BGC synteny blocks provide insight into the evolutionary relationships and diversity of BGCs within this complex group.

EsxA, a type VII secretion system-dependent effector, reveals a novel function in the sporulation of Bacillus cereus ATCC14579.

BACKGROUND: Bacillus cereus is a Gram-positive, spore-forming bacterium that produces a spectrum of effectors integral to bacterial niche adaptation and the development of various infections. Among those is EsxA, whose secretion depends on the EssC component of the type VII secretion system (T7SS). EsxA's roles within the bacterial cell are poorly understood, although postulations indicate that it may be involved in sporulation. However, the T7SS repertoire in B. cereus has not been reported, and its functions are unestablished. METHODS: We used the type strain, B. cereus ATCC14579, to generate ΔessC mutant through homologous recombination using the homing endonuclease I-SceI mediated markerless gene replacement. Comparatively, we analyzed the culture supernatant of type strain and the ΔessC mutant through Liquid chromatography-tandem mass spectrometry (LC-MS/MS). We further generated T7SSb-specific gene mutations to explore the housekeeping roles of the T7SSb-dependent effectors. The sporulation process of B. cereus ATCC14579 and its mutants was observed microscopically through the classic Schaeffer-Fulton staining method. The spore viability of each strain in this study was established by enumerating the colony-forming units on LB agar. RESULTS: Through LC-MS/MS, we identified a pair of nearly identical (94%) effector proteins named EsxA belonging to the sagEsxA-like subfamily of the WXG100 protein superfamily in the culture supernatant of the wild type and none in the ΔessC mutant. Homology analysis of the T7SSb gene cluster among B. cereus strains revealed diversity from the 3' end of essC, encoding additional substrates. Deletions in esxA1 and esxA2 neither altered cellular morphology nor growth rate, but the ΔesxA1ΔesxA2 deletion resulted in significantly fewer viable spores and an overall slower sporulation process. Within 24 h culture, more than 80% of wild-type cells formed endospores compared to less than 5% in the ΔesxA1ΔesxA2 mutant. The maximum spore ratios for the wild type and ΔesxA1ΔesxA2 were 0.96 and 0.72, respectively. Altogether, these results indicated that EsxA1 and EsxA2 work cooperatively and are required for sporulation in B. cereus ATCC14567. CONCLUSION: B. cereus ATCC14579 possesses two nearly identical T7SSb-dependent effectors belonging to the sagEsxA-like proteins. Simultaneous deletion of genes encoding these effectors significantly delayed and reduced sporulation, a novel finding for EsxA.

Phyto-safe in vitro regeneration and harnessing antimicrobial-resistant endophytes as bioinoculants for enhanced growth and secondary metabolites yield in Nilgirianthus ciliatus.

Nilgirianthus ciliatus, extensively exploited for its pharmacological properties, is now classified as vulnerable. In vitro micropropagation offers a sustainable approach for ecological conservation and rational utilization of this biodiversity resource. This study aimed to reduce endophytes during in vitro propagation and isolating antimicrobial-resistant endophytes from N. ciliatus by employing various concentrations and exposure times of Plant Preservative Mixture (PPM). Optimal results were observed when nodal explants treated with 0.3% PPM for 8 h, followed by inoculation in Murashige and Skoog (MS) medium supplemented with 3 mg/L 6-benzylaminopurine (BAP) and 0.3% PPM. This protocol achieved 82% shoot regeneration with minimal endophytic contamination, suggesting that the duration of explant exposure to PPM significantly influences endophyte reduction. Two antimicrobial-resistant endophytes were isolated and identified as Bacillus cereus and Acinetobacter pittii through 16S rDNA sequencing. These endophytes exhibited plant growth-promoting characteristics, including amylolytic, proteolytic, lipolytic activities, indole-3-acetic acid production, phosphate solubilization, and stress tolerance. In vivo application of these endophytes as bioinoculants to N. ciliatus not only improved growth parameters but also significantly increased the levels of pharmacologically important compounds, squalene, and stigmasterol, as confirmed by High-performance thin-layer chromatography (HPTLC). This study demonstrates that PPM is a promising alternative for sustainable micropropagation of N. ciliatus. Furthermore, it highlights the potential of antimicrobial-resistant endophytes as bioinoculants to improve growth and medicinal value, offering a sustainable solution for conservation and large-scale cultivation of this species.

An Outbreak of Bacillus cereus Emetic Toxin Mediated Food Poisoning After Consumption of Fried Rice in Assam.

Bacillus cereus is an emerging food-borne pathogen responsible for two types of food poisoning: emetic and diarrhoeal type. Here we report an emetic type of food-borne illness attributable to Bacillus cereus. On 2nd February, 2021, 202 people suffered from pain in abdomen and vomiting after consuming the rice provided during a public gathering in Diphu, Assam. Culture of leftover fried rice showed growth of Bacillus cereus group of organisms. Molecular detection of enterotoxin and emetic toxin genes was done in the isolated strains by polymerase chain reaction. Multi locus sequence typing (MLST) and phylogenetic analysis was done to characterise the isolated strains. A total of five strains of Bacillus cereus were isolated. Ces gene was found in isolates GMC22 & GMC24 and other enterotoxins producing genes were found in isolates GMC23 and GMC24. MLST identified four sequence types (STs) (ST1051, ST1616, ST998 and ST1000). Phylogenetic analysis clustered ST-1051 assigned to the GMC22 strain into the previously defined clade I and was in close relation with ST-144, representing a new cereulide-producing emetic cluster. As Bacillus cereus is a common contaminant of foods, it is essential to evaluate the pathogenic potential of the bacteria for a definite link between causative agents and the illness. MLST can be used to characterize the Bacillus cereus strains isolated from outbreak samples in order to link the probable pathogens with the illness. In this outbreak, we suggest that ST-1051 is the strain responsible for the food-borne illness, which was predominantly of emetic type.

Draft genome sequence of Bacillus cereus sequence type 2255 isolated from powdered infant formula.

The draft genome sequence of a Bacillus cereus strain, DFPST-SP1, isolated from powdered infant formula in the United States is reported. The 5,216,828-bp draft genome comprises 46 contigs with 66.4× coverage and 36% GC content and was typed as sequence type 2255.

Inhibitory effect and mechanism of violacein on planktonic growth, spore germination, biofilm formation and toxin production of Bacillus cereus and its application in grass carp preservation.

Bacillus cereus is a ubiquitous foodborne pathogen commonly found in various foods. Its ability to form spores, biofilms and diarrhoeal and/or emetic toxins further exacerbates the risk of food poisoning. Violacein is a tryptophan derivative with excellent antibacterial activity. However, the knowledge on the antibacterial action of violacein against B. cereus was lacking, and thus this study aimed to investigate the antibacterial activity and mechanism. The antibacterial results demonstrated that minimum inhibitory concentration and minimum bactericidal concentration of violacein were 3.125 mg/L and 12.50 mg/L, respectively. Violacein could effectively inhibit planktonic growth, spore germination and biofilm formation of B. cereus (P < 0.001). Meanwhile, violacein significantly downregulated the expression of toxin genes, including nheA (P < 0.05), nheB (P < 0.001), bceT (P < 0.01), cytK (P < 0.001), hblC (P < 0.001) and hblD (P < 0.001). Results of extracellular alkaline phosphatase, nucleotide and protein leakage assays and scanning and transmission electron microscopy observation tests showed violacein destroyed cell walls and membranes of B. cereus. In addition, 6.25 mg/kg of violacein could significantly inhibit B. cereus in grass carp fillets (P < 0.05). These results demonstrate that violacein has great potential as an effective natural antimicrobial preservative to control food contamination and poisoning events caused by B. cereus.

Characterization of toxigenic genes of Bacillus cereus strains isolated from different spices sold in Algeria.

Many cooked foods are prepared with spices and dried herbs; these can be contaminated by several types of microorganisms, including aerobic spore-forming bacteria. The Bacillus cereus group is very widespread in nature and is known among the common food contaminants. They are involved in food poisoning, causing two types of syndromes, diarrheal and emetic. The aims of the present work were to determine the prevalence of toxigenic Bacillus cereus spores in spices and herbs marketed in the Laghouat area and to identify their toxigenic genes via PCR. Among the 191 samples, 14.13% were determined to be B. cereus, with concentrations ranging from 2.52 to 5.82 log cfu/g, where the highest level of contamination was observed in allspice and ginger. Moreover, entFM (100%), nhe (88.23%) and cytK (70.58%) were the most frequently identified toxin genes, whereas hbl (23.52%) was less common, and no emetic toxin-encoding gene (cesB) was found in any of the samples. Considering the results of the present study, the B. cereus microbial load and toxin gene profiles of spices show that spices have potential for public health in Algeria. In this context, it is crucial to guarantee the microbiological safety of spices by respecting good hygiene practices, eliminating bacterial spores and toxin production via sterilization and using appropriate packaging for these products.

Structural insight into sugar-binding modes of microbial ß-amylase.

ß-Amylase, which catalyses the release of ß-anomeric maltose from the non-reducing end of starch, is widely used in the food industry. Increasing its enzyme activity through protein engineering might improve the efficiency of food processing. To obtain detailed structural information to assist rationale design, here the crystal structure of Bacillus cereus ß-amylase (BCB) complexed with maltose was determined by molecular replacement and refined using anisotropic temperature factors to 1.26 Å resolution with Rwork/Rfree factors of 12.4/15.7 %. The structure contains six maltose and one glucose molecules, of which two maltose and one glucose are bound at sites not previously observed in BCB structures. These three new sugar-binding sites are located on the surface and likely to be important in enhancing the degradation of raw-starch granules. In the active site of BCB, two maltose molecules are bound in tandem at subsites -2 âˆ¼ -1 and +1 âˆ¼ +2. Notably, the conformation of the glucose moiety bound at subsite -1 is a mixture of α-anomeric distorted 1,4B boat and 4C1 chair forms, while those at subsites -2, +1 âˆ¼ +2 are all in the 4C1 chair forms. The O1 of the distorted α-glucose residue at subsite -1 occupies the position of the putative catalytic water, forming a hydrogen bond with OE1 of Glu367 (base catalyst), suggesting that this distorted sugar is not involved in catalysis. Together, these findings pave the way for further improving the functionality of microbial ß-amylase enzymes.

Antimicrobial and Phylogenomic Characterization of Bacillus cereus Group Strains Isolated from Different Food Sources in Italy.

Background:Bacillus cereus is a widespread environmental Gram-positive bacterium which is especially common in soil and dust. It produces two types of toxins that cause vomiting and diarrhea. At present, foodborne outbreaks due to Bacillus cereus group bacteria (especially Bacillus cereus sensu stricto) are rising, representing a serious problem in the agri-food supply chain. Methods: In this work, we analyzed 118 strains belonging to the Bacillus cereus group, isolated from several food sources, for which in vitro and in silico antibiotic resistance assessments were performed. Results: Many strains showed intermediate susceptibility to clindamycin, erythromycin, and tetracycline, suggesting an evolving acquisition of resistance against these antibiotics. Moreover, one strain showed intermediate resistance to meropenem, an antibiotic currently used to treat infections caused by Bacillus cereus. In addition to the phenotypic antimicrobial resistance profile, all strains were screened for the presence/absence of antimicrobial genes via whole-genome sequencing. There was inconsistency between the in vitro and in silico analyses, such as in the case of vancomycin, for which different isolates harbored resistance genes but, phenotypically, the same strains were sensitive. Conclusions: This would suggest that antibiotic resistance is a complex phenomenon due to a variety of genetic, epigenetic, and biochemical mechanisms.

Genomic Insights into the Pathogenicity and Drug-Resistance of a Bacillus cereus Isolated from Human Teeth.

Background: Bacillus cereus is a common bacterium found in the environment. Some strains can cause food poisoning, and very few can cause clinically severe infections, leading to death. Here, we characterized the genome sequence of B. cereus LIN78 isolated from teeth with deep caries and compared it with those of 25 other related species. Methods: Third-generation sequencing technology, bacteriological analyses, biochemistry, and mass spectrometry were applied to characterize the drug-resistance genes and virulence factors of B. cereus LIN78. Results: The complete genome sequence of B. cereus Lin78 consists of 5647 genes distributed on a circular chromosome, a 393 kbp plasmid, and 928 pseudogenes (37.4% of whole-genome DNA). The LIN78 genome contains 14 sets of 16s, 23s, and 5s ribosomal RNA operons; 106 tRNA genes, one tmRNA, 12 genomic islands, six prophases, 64 repeats; 37 antibiotic-resistant genes; and 1119 putative virulence genes, including enterotoxins and cytolysins. The B. cereus LIN78 genome carries multiple copies of non-ribosomal polypeptide synthetase (NRPS) and post-translationally modified peptides (RiPPs). Phylogenetic analysis of the 26 B. cereus strains showed that B. cereus LIN78 is evolutionarily closely related to B. thuringiensis ATCC 10792 and B. cereus ATCC 14579. Conclusion: The newly isolated B. cereus carries many virulence genes, including enterotoxins and hemolysins, similar to B. anthracis, and multiple antibiotic resistance genes. These findings suggest that the strain has a potential risk of causing disease. Our studies are vital for further exploration of the evolution of B. cereus, its pathogenic mechanisms, and the control and treatment of bacterial infections.

The microbial contaminants of plant-based meat analogues from the retail market.

The aim of the study was to analyse the key microbial contaminants of plant-based meat analogues (PBMA) from retail. A total of 43 samples of PBMAs (12 frozen/31 chilled) in the "ready-to-cook" category, such as hamburgers, meatballs or breaded imitation steaks were purchased in retail stores in the Czech Republic in summer (n = 21) and autumn 2022 (n = 22). The detected indicator bacteria (total viable count, lactic acid bacteria, Enterobacteriaceae, yeasts, moulds) had relatively low values in the analysed PBMA samples and only rarely reached levels of 7 log CFU/g. E. coli, STEC and coagulase-positive staphylococci were not detected by isolation from plates in any of analysed samples. Mannitol positive Bacillus spp. were isolated from almost half of the analysed samples of the PBMA. B. cereus sensu lato was isolated from 3 samples by isolation from plates, and after enrichment in 35 samples (81 %). Clostridium perfringens could not be detected by isolation from plates, nevertheless after multiplication, it was detected in 21 % of samples. Analyses of PBMA samples revealed considerable variability in microbial quality. The presence of spore-forming bacteria with the potential to cause foodborne diseases is alarming. However, to evaluate the risks, further research focused on the possibilities of growth under different conditions of culinary treatment and preservation is needed.

Phytochemical Characteristics and Antimicrobial Effects of Nonea rossica (Boraginaceae) Extracts.

Phytochemical characteristics and antimicrobial properties of extracts were studied in Nonea rossica Steven (Boraginaceae), which is widespread in Russia. The aerial part (herb) of N. rossica was harvested from a steppe meadow in the Novosibirsk region during flowering. The qualitative composition of biologically active compounds (BACs) was determined by thin-layer chromatography. Quantitative assays were carried out by spectrophotometry; flavonoids, hydroxycinnamic acids, and coumarin-like compounds were measured with reference to rutin, caffeic acid, and coumarin, respectively. Antimicrobial activity was determined by the serial dilution method. Gram-positive bacterial (Staphylococcus aureus ATCC 6538 FDA 209P and Bacillus cereus ATCC 10702) and fungal (Candida albicans NCTC 885-653) strains were used as test cultures. Phenolic BACs (hydroxycinnamic acids, flavonoids, and coumarins) were detected, and their quantitative contents determined. The highest yield of phenolic BACs was achieved using 40-70% ethanol as an extractant. Antimicrobial activity against S. aureus and B. cereus and antifungal activity against C. albicans were detected in N. rossica herb extracts prepared using 40-70% ethanol. The extracts were tested for the contents of caffeic acid and coumarin. Synergistic interactions of these compounds determined the bactericidal and fungistatic properties of the extracts.

Bioemulsifier from sponge-associated bacteria reduces staphylococcal biofilm.

Biofilm formation is a major health concern and studies have been pursued to find compounds able to prevent biofilm establishment and remove pre-existing biofilms. While biosurfactants (BS) have been well-known for possessing antibiofilm activities, bioemulsifiers (BE) are still scarcely explored for this purpose. The present study aimed to evaluate the bioemulsifying properties of cell-free supernatants produced by Bacillaceae and Vibrio strains isolated from marine sponges and investigate their antiadhesive and antibiofilm activities against different pathogenic Gram-positive and Gram-negative bacteria. The BE production by the marine strains was confirmed by the emulsion test, drop-collapsing, oil-displacement, cell hydrophobicity and hemolysis assays. Notably, Bacillus cereus 64BHI1101 displayed remarkable emulsifying activity and the ultrastructure analysis of its BE extract (BE64-1) revealed the presence of structures typically observed in macromolecules composed of polysaccharides and proteins. BE64-1 showed notable antiadhesive and antibiofilm activities against Staphylococcus aureus, with a reduction of adherence of up to 100 % and a dispersion of biofilm of 80 %, without affecting its growth. BE64-1 also showed inhibition of Staphylococcus epidermidis and Escherichia coli biofilm formation and adhesion. Thus, this study provides a starting point for exploring the antiadhesive and antibiofilm activities of BE from sponge-associated bacteria, which could serve as a valuable tool for future research to combat S. aureus biofilms.

The mitigating effects and mechanisms of Bacillus cereus on chronic cadmium poisoning in Litopenaeus vannamei based on histopathological, transcriptomic, and metabolomic analyses.

Shrimp are non-negligible victims of cadmium (Cd) contamination, and there is still a lack of strategies for mitigating Cd toxicity in shrimp. Bacillus cereus, with its significant heavy metal (HM) tolerance and chelating effects, is a representative beneficial bacterium to be investigated for mitigating the toxicity of Cd exposure. This study revealed the effects and potential mechanisms of B. cereus in mitigating chronic Cd toxicity in shrimp by analyzing growth performance, hepatopancreatic Cd accumulation, pathology, as well as comprehensive hepatopancreatic transcriptomics and metabolomics in Litopenaeus vannamei. The results showed that shrimp's growth inhibition, hepatopancreatic Cd accumulation and physiological structure damage in B. cereus+chronic Cd group were effectively alleviated compared with the chronic Cd treatment group. The pathways related to amino acid metabolism, glycolipid metabolism, immune response, and antioxidant stress were significantly activated in the B. cereus+chronic Cd group, including glycolysis, pentose phosphate pathway, oxidative phosphorylation, biosynthesis of amino acids, and biosynthesis of unsaturated fatty acids pathways. The key differentially expressed genes (e.g., macrophage migration inhibitory factor, glycine cleavage system H protein, glycine dehydrogenase, phosphoglucomutase-2, asparaginase, ATP synthase subunit, cytochrome c, and 4-hydroxyphenylpyruvate dioxygenase) and metabolites (e.g., L-leucine, D-ribose, gluconic acid, 6-Phosphogluconic acid, sedoheptulose 7-phosphate, 1-Kestose, glyceric acid, arachidic acid, prostaglandins, 12-Keto-tetrahydro-leukotriene B4, and gamma-glutamylcysteine) associated with the above pathways were significantly altered. This study demonstrated that B. cereus is an effective mitigator for the treatment of chronic Cd poisoning in shrimp. B. cereus may play a role in alleviating the toxicity of Cd by enhancing the antioxidant performance, immune defense ability, metabolic stability, and energy demand regulation of shrimp. The study provides reference materials for the study of B. cereus in alleviating Cd toxicity of shrimp and broadens the application of probiotics in treating HM toxicity.