Assessment of antimicrobial activity and GC-MS using culture filtrate of local marine Bacillus strains.

Secondary metabolites produced by Bacillus species from marine sources encompass a variety of compounds such as lipopeptides, isocoumarins, polyketides, macrolactones, polypeptides and fatty acids. These bioactive substances exhibit various biological activities, including antibiotic, antifungal, antiviral, and antitumor properties. This study aimed to isolate and identify a particular species of Bacillus from marine water and organisms that can produce bioactive secondary metabolites. Among the 73 Bacillus isolates collected, only 5 exhibited antagonistic activity against various viral and bacterial pathogens. The active isolates were subjected to 16S rRNA sequencing to determine their taxonomical affiliation. Among them, Bacillus tequilensis CCASU-2024-66 strain no. 42, with the accession number ON 054302 in GenBank, exhibited the highest inhibitory potential. It displayed an inhibition zone of 21 mm against Bacillus cereus while showing a minimum zone of inhibition of 9 mm against Escherichia coli and gave different inhibition against pathogenic fungi, the highest inhibition zone 15 mm against Candida albicans but the lowest inhibition zone 10 mm was against Botrytis cinerea, Fusarium oxysporum. Furthermore, it demonstrated the highest percentage of virucidal effect against the Newcastle virus and influenza virus, with rates of 98.6% and 98.1%, respectively. Furthermore, GC-MS analysis was employed to examine the bioactive substance components, specifically focusing on volatile and polysaccharide compounds. Based on these results, Bacillus tequilensis strain 42 may have the potential to be employed as an antiviral agent in poultry cultures to combat Newcastle and influenza, two extremely destructive viruses, thus reducing economic losses in the poultry production sector. Bacteria can be harnessed for the purpose of preserving food and controlling pathogenic fungi in both human and plant environments. Molecular docking for the three highly active derivatives 2,3-Butanediol, 2TMS, D-Xylopyranose, 4TMS, and Glucofuranoside, methyl 2,3,5,6-tetrakis-O-(trimethylsilyl) was carried out against the active sites of Bacillus cereus, Listeria monocytogenes, Candida albicans, Newcastle virus and influenza virus. The data obtained from molecular docking is highly correlated with that obtained from biology. Moreover, these highly active compounds exhibited excellent proposed ADMET profile.

An Anti-MRSA Phage From Raw Fish Rinse: Stability Evaluation and Production Optimization.

Accumulating evidence has denoted the danger of resistance in tenacious organisms like methicillin-resistant Staphylococcus aureus (MRSA). MRSA, a supple bacterium that adopts a variety of antibiotic resistance mechanisms, is the cause of multiple life-threatening conditions. Approaching a post-antibiotic era, bacteria-specific natural predators, bacteriophages, are now given the chance to prove eligible for joining the antibacterial weaponry. Considering the foregoing, this study aimed at isolating bacteriophages with promising anti-MRSA lytic activity, followed by characterization and optimization of the production of the bacteriophage with the broadest host range. Five phages were isolated from different environmental sources including the rinse of raw chicken egg, raw milk, and, remarkably, the raw meat rinses of chicken and fish. Examined for lytic activity against a set of 23 MRSA isolates collected from various clinical specimens, all five phages showed relatively broad host ranges with the bacteriophage originally isolated from raw fish rinse showing lytic activity against all the isolates tested. This phage is suggested to be a member of Siphoviridae family, order Caudovirales, as revealed by electron microscopy. It also exhibited good thermal stability and viability at different pH grades. Moreover, it showed reasonable stability against UV light and all viricidal organic solvents tested. Optimization using D-optimal design by response surface methodology was carried out to enhance the phage yield. The optimum conditions suggested by the generated model were a pH value of 7, a carbon source of 0.5% w/v sucrose, and a nitrogen source of 0.1% w/v peptone, at a temperature of 28°C and a bacterial inoculum size of 107 CFU/ml, resulting in a 2 log-fold increase in the produced bacteriophage titer. Overall, the above findings indicate the lytic ability inflicted by this virus on MRSA. Apparently, its stability under some of the extreme conditions tested implies its potential to be a candidate for pharmaceutical formulation as an anti-MRSA therapeutic tool. We hope that bacteriophages could tip the balance in favor of the human front in their battle against multidrug-resistant pathogens.

Morphogenesis and Genetic Diversity of Some Virulent Phages Specific for Bacillus velezensis.

<b>Background and Objective:</b> Phosphorus (P) is one of the most limiting nutrients for plant growth. Phosphorus deficiency is limiting crop production in many agricultural soils worldwide. The application of phosphorus solubilizing bacteria (PSB) to soils can replace or partially reduce using of inorganic P fertilizers. A bacteriophage, or phage, is a virus that infects a bacterial cell, taking over the host cell's genetic material. The four phages were propagated, purified, studied for the morphological properties, finally studying the genetic diversity. <b>Materials and Methods:</b> Obtained, examined the efficiency and identification of bacteria for solubilizing phosphorus. Isolation, studying the properties and studying genetic diversity. <b>Results:</b> Four virulent phages (Bv₁, Bv₂, Bv₃ and Bv₄) specific for <i>Bacillus velezensis</i> were isolated from the Egyptian soil. The <i>Bacillus</i> phages were purified by alternative low and high-speed centrifugation methods. Electron micrographs showed that phages appeared to be a member of the <i>Siphoviridae </i>family based on their structure and particle morphology (the particles have a head and long non-contractile tail). Sodium Dodecyl Sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) technique was performed to determine the properties of viral proteins. It was found that the Bv₁ virus had five structural proteins, while Bv₂ and Bv₃ virus had eight structural proteins and finally, the Bv₄ virus had ten structural proteins. The purity and quantity of isolated DNAs were determined spectrophotometrically. Data showed that the concentration of Bv₁ DNA was 0.75 µg, Bv₂ DNA and Bv₃ DNA was 0.60 µg and finally Bv₄ DNA 0.55 µg µL¹. The analysis of genetic material of <i>B. velezensis</i> phages was determined based on both the ISSR-PCR technique and the effect of restriction enzymes. Data showed different amplification patterns with all phages. <b>Conclusion:</b> The bacteriophages of <i>B. velezensis</i> were isolated from soil, propagated, purified, study some of its properties.

Isolation and Characterization of Antagonistic Activity of Some Streptomycetes and their Specific Actinophages from Soil.

<b>Background and Objective:</b> Phages specific to actinomycetes are common, active in the soil and gladly detected. Soil streptomycetes are having antibiosis activities against numerous bacteria, fungi and plant viruses. Thus, this study was designed to isolate, purify and characterize some streptomycetes active against some microorganisms from soil followed by isolation of their specific phages. <b>Materials and Methods:</b> Antagonistic activities of these streptomycetes isolates were tested against <i>Bacillus subtilis</i>,<i> Pseudomonas</i> sp., <i>Serratia</i> sp. and <i>Aspergillus niger</i>. To confirm their biological characterization of the streptomycetes isolates under investigation, the 16SrRNA gene was also used. The presence of specific lysate actinophages in the soil samples were tested by spot test technique and then propagated and purified for further characterization. The morphology of the purified actinophages was determined by electron microscopy. <b>Results:</b> The five selected <i>Streptomyces</i> isolates having effective antagonistic activity were biologically and molecularly identified as <i>Streptomyces sclerogranulatus </i>(QQ06), <i>Streptomyces mutabilis </i>(QQ07), <i>Streptomyces heilongjiangensis </i>(QQ08), <i>Streptomyces sparsus </i>(QQ09) and <i>Streptomyces purpurascens </i>(QQ10) strains. Electron micrographs showed the presence of filamentous virus-like particles with lengths of 21.4×928.57, 25×750, 21.4×857.14, 21.4×885.7 and 21.4×857.14 nm specific to <i>Streptomyces</i> strains QQ06, QQ07, QQ08, QQ09 and QQ10, respectively and belong to the family Inoviridae. <b>Conclusion:</b> Phage of Inoviridae was considered as the first time against streptomycetes isolates, therefore, additional and advanced studies should be carried out at the level of molecular characterization.

Probiotic Lactobacillus and Bifidobacterium strains possess safety characteristics, antiviral activities and host adherence factors revealed by genome mining.

BACKGROUND: Probiotics belonging to Lactobacillus and Bifidobacterium spp. have been exploited for their health benefits in treatment and prevention of many pathological conditions and promoting human health. Recent advances in understanding probiotics-human interaction through microbiome research in the context of various medical conditions suggest their provisional role in preventive, personalized, and predictive medicine. To streamline their application in disease prevention, development of personalized-based treatments, or their use as biomarkers for predictive diagnosis, in vitro screening for strains with potential probiotic properties should be performed. In this work, we aimed to emphasize the probiotic features of four Lactobacillus and two Bifidobacterium probiotic strains which showed antagonistic properties against microbial pathogens. METHODS: Firstly, cytotoxicity assessment of cell-free preparations from these strains was performed using a baby hamster kidney (BHK) cells and cell viability was measured by means of sulfo-rhodamine B stain. Secondly, Newcastle disease (ND) and infectious bursal disease (IBD) viruses which pose a great threat in infected poultry were used for assessing antiviral activity of probiotics. Thirdly, the genomes of six probiotic strains were used to identify genes encoding host adherence factors that mediate interaction with human tissues. RESULTS: Probiotic preparations exhibited insignificant toxicity as indicated by the high survival rate of BHK cells (surviving fraction varied from 0.82 to 0.99) as compared to the untreated control. Cell-free preparations of probiotics mixed with equal volume of ND and IBD viruses (106 and 104 Tissue Culture Infectious Dose 50, respectively) reduced the titer of ND and IBD viruses on chicken embryo fibroblast cells. Genome mining analysis revealed that the draft genomes of these strains were predicted to encode LPXTG-containing proteins, surface layer proteins, tight adherence pili, sortase-dependent pili, fibronectin, or collagen binding proteins and other factors that adhere to human tissues such as mucus. Such adherence factors enable probiotic bacteria to interact and colonize the host. CONCLUSION: Taken together, safety privileges, antiviral activities, and genomically encoded host interaction factors confirmed probiotic features of the six probiotic strains and their potential in promoting human health.