Diversity, molecular phylogenetics, and antibiotic biosynthetic potential of endophytic Actinobacteria isolated from medicinal plants in Nigeria.

Actinobacteria that are found in nature have enormous promise for the growth of the pharmaceutical sector. There is a scarce report on the antimicrobial activities of endophytic Actinobacteria from Nigeria. As a result, this study evaluated the Actinobacteria isolated from Nigerian medicinal plants in terms of their biodiversity, phylogenetics, and ability to produce antimicrobial compounds. Following accepted practices, Actinobacteria were isolated from the surface-sterilized plant parts. They were identified using 16S rRNA sequencing, microscopic, and morphological methods. The cell-free broth of Actinobacteria isolates was subjected to antimicrobial assay by agar well diffusion. Molecular evolutionary and genetic analysis (MEGA) version X was used for phylogenetic analysis, and the interactive tree of life (iTOL) version 6.0 was used to view the neighbour-joining method-drawn tree. A total of 13 Actinobacteria were recovered, belonging to three genera including 10 strains of Streptomyces, 2 strains of Saccharomonospora, and only 1 strain of Saccharopolyspora. They showed inhibitory activity against several bacterial pathogens. The phylogenetic tree generated from the sequences showed that our isolates are divergent and distinct from other closely related strains on the database. Further, optimization of the antibiotic production by selected Saccharomonospora sp. PNSac2 was conducted. It showed that the optimal conditions were the ISP2 medium (1-2% w/v salt) adjusted to pH of 8 at 30-32℃ for 12-14 days. In conclusion, endophytic Actinobacteria dwelling in Nigerian soils could be a promising source of new antibiotics. Future research is warranted because more genomic analysis and characterization of their metabolites could lead to the development of new antibacterial medicines.

Phylogenetic insights and antimicrobial biosynthetic potential of Serratia sp. XAFb12 and Pseudomonas sp. XAFb13 from Xylopia aethiopica.

The extensive distribution of Xylopia aethiopica across the continent of Africa has firmly established its medicinal value in diverse disease management. While its phytochemistry is well established, the diversity, molecular, biochemical, and antimicrobial-biosynthetic characterizations of culturable bacterial endophytes residing in fruits of X. aethiopica have not been studied previously. Additionally, danger continues to loom the global health care and management due to antibiotic resistance; hence, the discovery of microbial natural products especially from endophytes could offer a lasting solution to the quest for novel antimicrobial compounds. In this study, we isolated two bacterial endophytes Serratia sp. XAFb12 and Pseudomonas sp. XAFb13 from fresh X. aethiopica fruit. The 16S rRNA gene sequencing, Vitex biochemical test, Gram staining, and 16S rRNA gene analysis were used to confirm their phenotypic and genotypic profiles. Phylogenetic tree analysis reveals their divergence in a separate branch, indicating their uniqueness. The crude extract of both strains showed inhibition against all tested bacterial and fungal pathogens. The minimum inhibition concentration (MIC) ranged from 2.5 to 10%. Chemical analysis of the crude extracts using gas chromatography-mass spectroscopy (GC-MS) revealed the most abundant compounds to be hydrocinnamic acid, 2-piperidinone, 5-isopropylidene-3,3-dimethyl-dihydrofuran-2-one, and diethyl trisulfide. The bacterial endophytes linked to X. aethiopica were described in this study for the first time in relation to clinically significant pathogens. Our findings imply that crude extracts of the endophytic bacteria from X. aethiopica could be potentially employed as antibiotics. Hence, it is crucial to characterize the active ingredient in further detail for future pharmaceutical applications.

Mining metagenomes reveals diverse antibiotic biosynthetic genes in uncultured microbial communities.

Pathogens resistant to antimicrobials form a significant threat to public health worldwide. Tackling multidrug-resistant pathogens via screening metagenomic libraries has become a common approach for the discovery of new antibiotics from uncultured microorganisms. This study focuses on capturing nonribosomal peptide synthase (NRPS) gene clusters implicated in the synthesis of many natural compounds of industrial relevance. A NRPS PCR assay was used to screen 2976 Escherichia coli clones in a soil metagenomic library to target NRPS genes. DNA extracts from 4 clones were sequenced and subjected to bioinformatic analysis to identify NRPS domains, their phylogeny, and substrate specificity.Successfully, 17 NRPS-positive hits with a biosynthetic potential were identified. DNA sequencing and BLAST analysis confirmed that NRPS protein sequences shared similarities with members of the genus Delftia in the Proteobacteria taxonomic position. Multiple alignment and phylogenetic analysis demonstrated that clones no. 15cd35 and 15cd37 shared low bootstrap values (54%) and were distantly far from close phylogenetic neighbors. Additionally, NRPS domain substrate specificity has no hits with the known ones; hence, they are more likely to use different substrates to produce new diverse antimicrobials. Further analysis confirmed that the NRPS hits resemble several transposon elements from other bacterial taxa, confirming its diversity. We confirmed that the analyses of the soil metagenomic library revealed a diverse set of NRPS related to the genus Delftia. An in-depth understanding of those positive NRPS hits is a crucial step for genetic manipulation of NRPS, shedding light on alternative novel antimicrobial compounds that can be used in drug discovery and hence supports the pharmaceutical sector.

Synergistic inhibition of Pseudomonas fluorescens growth and proteases activities via sodium chlorite-based oxyhalogen.

Pseudomonas fluorescens is considered among the main spoilage microorganisms due to its ability to produce proteases. Food deterioration caused by spoilage microorganisms has a major impact on food quality and the environment. The inactivation of Pseudomonas fluorescens growth and protease production was intensively investigated with the use of Salmide®, A Sodium Chlorite-Based Oxy-halogen Disinfectant. A unique M9 media was also developed to assure sufficient protease productions with different mutants of Pseudomonas fluorescens as a microbioreactor. Mutations were induced by classical whole-cell mutagenesis using N-methyl-N'- nitro-N-nitrosoguanidine (NTG). A dramatic decrease occurred in protease activity when different Salmide concentrations (5, 10, and 15 ppm) were added to the growth culture followed by a complete inhibition concentration (20, 25, 50, and 100 ppm) of Salmide. However, no significant inhibition occurred once it is secreted out of cells. Some mutants were resistant and remains highly stable with high protease production under stressful conditions of Sodium Chlorite-Based Oxy-halogen. The production of the protease showed a linear correlation with the increase in incubation time using a continuous culture bioreactor system and recorded maximum protease activity after 40 h. Our findings would offer alternative antimicrobial procedures for food and industrial sectors.

Consolidating food safety measures against COVID-19.

BACKGROUND: The world is facing an extraordinarily unprecedented threat from the COVID-19 pandemic triggered by the SARS-CoV-2 virus. Global life has turned upside down, and that several countries closed their borders, simultaneously with the blockage of life cycle as a result of the shutdown of the majority of workplaces except the food stores and some few industries. MAIN BODY: In this review, we are casting light on the nature of COVID-19 infection and spread, the persistence of SARS-CoV-2 virus in food products, and revealing the threats arising from the transmission of COVID-19 in food environment between stakeholders and even customers. Furthermore, we are exploring and identifying some practical aspects that must be followed to minimize infection and maintain a safe food environment. We also present and discuss some World Health Organization (WHO) guidelines-based regulations in food safety codes, destined to sustain the health safety of all professionals working in the food industry under this current pandemic. CONCLUSION: The information compiled in this manuscript is supporting and consolidating the safety attributes in food environment, for a prospective positive impact on consumer confidence in food safety and the citizens' public health in society. Some research is suggested on evaluating the use and potentiality of native and chemical modified basic proteins as possible practices aiming at protecting food from bacterial and viral contamination including COVID-19.

Antimicrobial Profile of Actinomycin D Analogs Secreted by Egyptian Desert Streptomyces sp. DH7.

Egyptian deserts are an underexplored ecological niche, especially the Sinai Peninsula. In our recent study, we explored this extreme environment and shed light on the bioactive capabilities of desert Actinobacteria isolated from Sinai. Fifty desert Actinobacteria were isolated from the Sinai desert using mineral salt media, basal media, and starch casein media. The filtrate of Streptomyces sp. DH 7 displayed a high inhibitory effect against multidrug-resistant Staphylococcus aureus (MRSA) strains. The 16S rDNA sequencing confirmed that isolate DH7 belongs to the genus Streptomyces. The NJ phylogenetic tree showed relatedness to the Streptomyces flavofuscus strain NRRL B-2594 and Streptomyces pratensis strain ch24. The minimum inhibitory concentrations against MRSA were 16 and 32 µg/µL. Chemical investigation of the ethyl acetate extract of Streptomyces sp. DH7 led to the isolation and purification of natural products 1-4. Structure elucidation of the purified compounds was performed using detailed spectroscopic analysis including 1 and 2D NMR, and ESI-MS spectrometry. To the best of our knowledge, this is the first report for the isolation of compounds 1-4 from a natural source, while synthetic analogs were previously reported in the literature. Compounds 3-4 were identified as actinomycin D analogues and this is the first report for the production of actinomycin D analogs from the Sinai desert with an inhibitory effect against MRSA. We indorse further study for this analog that can develop enhanced antimicrobial activities. We confirm that the desert ecosystems in Egypt are rich sources of antibiotic-producing Actinobacteria.

Endophytic microbes from Nigerian ethnomedicinal plants: a potential source for bioactive secondary metabolites-a review.

BACKGROUND: Endophytes are highly beneficial species of microbes that live in symbiosis with plant tissues in the setting. Endophytes are difficult to isolate in their natural environment, and they are understudied despite being a rich source of bioactive molecules. There are varieties of new infectious diseases emerging across the world, necessitating a constant and expanded search for newer and more efficient bioactive molecules. Nigeria is known for its biodiversity in ethnomedicinal plants, yet these plants are understudied for endophytic microbes harbouring novel bioactive molecules. MAIN BODY: Endophytes are a source of novel organic natural molecules and are thought to be drug discovery frontiers. Endophyte research has contributed to the discovery of possible anticancer agents following the discovery of taxol. Endophyte research has contributed to the discovery of possible drug compounds with antimicrobial, antioxidant, antiviral, antidiabetic, anti-Alzheimers disease and immunosuppressive properties among others. These breakthroughs provide hope for combating incurable diseases, drug resistance, the emergence of new infectious diseases, and other human health issues. Finding new medicines that may be effective candidates for treating newly emerging diseases in humans has a lot of promise. Most studies have been on fungi endophytes, with just a few reports on bacterial endophytes. The biology of endophytic bacteria and fungi, as well as endophytic microbes isolated from Nigerian medicinal plants, their isolation methods, identification by morphological and molecular methods, fermentation, purification, identification of bioactive compounds and biosynthetic gene clusters are all covered in this study. CONCLUSION: In Nigeria, the sourcing and isolation of endophytes harboring biosynthetic gene clusters are still understudied, necessitating a rigorous quest for bioactive molecules in endophytes inhabiting various ethnomedicinal plants.

Novel Molecular Markers Linked to Pseudomonas aeruginosa Epidemic High-Risk Clones.

The population structure of Pseudomonas aeruginosa is panmictic-epidemic in nature, with the prevalence of some high-risk clones. These clones are often linked to virulence, antibiotic resistance, and more morbidity. The clonal success of these lineages has been linked to acquisition and spread of mobile genetic elements. The main aim of the study was to explore other molecular markers that explain their global success. A comprehensive set of 528 completely sequenced P. aeruginosa genomes was analyzed. The population structure was examined using Multilocus Sequence Typing (MLST). Strain relationships analysis and diversity analysis were performed using the geoBURST Full Minimum Spanning Tree (MST) algorithm and hierarchical clustering. A phylogenetic tree was constructed using the Unweighted Pair Group Method with Arithmetic mean (UPGMA) algorithm. A panel of previously investigated resistance markers were examined for their link to high-risk clones. A novel panel of molecular markers has been identified in relation to risky clones including armR, ampR, nalC, nalD, mexZ, mexS, gyrAT83I, gyrAD87N, nalCE153Q, nalCS46A, parCS87W, parCS87L, ampRG283E, ampRM288R, pmrALeu71Arg, pmrBGly423Cys, nuoGA890T, pstBE89Q, phoQY85F, arnAA170T, arnDG206C, and gidBE186A. In addition to mobile genetic elements, chromosomal variants in membrane proteins and efflux pump regulators can play an important role in the success of high-risk clones. Finding risk-associated markers during molecular surveillance necessitates applying more infection-control precautions.