Bacterial strains were isolated from different locations and screened for plant growth promoting (PGP) features. Comparative analyses of the 16S gene sequences of 5 of the strains indicated taxonomic relatedness within the genus Paenibacillus. A polyphasic taxonomic approach was employed to study the strains in detail to clarify their phylogenetic position. Genome-based analyses, including digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI), revealed values consistently below the accepted species delineation thresholds when compared with closest relatives of each strain. Additionally, the strains showed clear differences in their physiological and biochemical profiles to the type strains of the closest related species. A notably diverse set of genes potentially involved in plant growth promotion was detected in all strains. With respect to the analyses reported here, the following new names are proposed: Paenibacillus corni sp. nov., with AK-167T as the type strain (= LMG 34403T = DSM 121689T); Paenibacillus vaccinii sp. nov., with AK-264T as the type strain (= LMG 34404T = DSM 121714T); Paenibacillus ericacearum sp. nov., with AK-265T as the type strain (= CCM 6913T = LMG 34405T = DSM 121715T); Paenibacillus polytrichii sp. nov., with AK-286T as the type strain (= CCM 9614T = LMG 34406T = DSM 121716T); and Paenibacillus artemisiae sp. nov., with DT-106T as the type strain (= LMG 34409T = CCM 9610T = DSM 121690T).

Several bacterial strains affiliated with the genera Pseudomonas and Zestomonas were isolated from rhizosphere samples and screened for plant growth-promoting (PGP) traits. Nine strains were analyzed polyphasically, showing ANI and dDDH values below or near species thresholds, together with phenotypic and biochemical traits supporting their differentiation. Genomic analyses revealed a repertoire of PGP-associated functions, including vitamin and cofactor biosynthesis (riboflavin, cobalamin, and thiamin), and high-affinity nutrient acquisition systems as phosphate transporters and phosphonate utilization. All strains encoded traits relevant to the rhizosphere inferred from genome annotation, including genes involved in auxin and cytokinin biosynthesis, oxidative stress tolerance, dissimilatory nitrate reduction, sulfur assimilation, and siderophore production. Metagenomic screening showed that most species are globally distributed across plant-associated, soil, freshwater, and animal-associated habitats. Based on phylogenetic, genomic, and phenotypic evidence, the strains represent seven novel species: Pseudomonas rosaeacicularis sp. nov., with AK-381T as the type strain (= LMG 34445T = CCM 9596T); Pseudomonas corni sp. nov., with AK-10T as the type strain (= CCM 9599T = LMG 34325T); Pseudomonas oplopanacis sp. nov., with AK-188T as the type strain (= CCM 9593T = LMG 34326T); Pseudomonas salicis sp. nov., with AK-309T as the type strain (= CCM 9595T = LMG 34328T); Pseudomonas artemisiae sp. nov., with DT-100T as the type strain (= LMG 32880T = DSM 115114T = CCM 9281T); Pseudomonas imperatae sp. nov., with ST-212T as the type strain (CCM 9594T = LMG 34330T); and Zestomonas ipomoeae sp. nov., with ST-55T as the type strain (LMG 32881T = CCM 9283T = DSM 115239T).

Two novel bacterial strains, designated MP11MiT and MP213FoT, were isolated from the burying beetle Nicrophorus vespilloides and subjected to a polyphasic taxonomic study. Strain MP11MiT was placed in the genus Gordonia, showing the highest 16S rRNA gene sequence similarities to Gordonia liuliyuniae HY366T (98.8% sequence identity), followed by Gordonia zhenghanii HY002T (98.4%). The strain exhibited low genomic relatedness to its closest relatives [ANI (average nucleotide identity) <90% and dDDH (digital DNA-DNA hybridization) <37%], far below the accepted species thresholds (95-96% ANI and 70% dDDH), unambiguously supporting species-level distinctiveness. Strain MP213FoT was affiliated with the genus Pseudochrobactrum, with Pseudochrobactrum algeriense LMG 32378T as its closest relative. ANI (79.4-79.9%), dDDH (22.8-23.1%), average amino acid identity (82.5-83.8%) and percentage of conserved proteins (71.6-77.2%) values confirmed its clear separation from all hitherto recognized species of this genus. The genome of MP213FoT comprises a circular chromosome and a plasmid carrying conserved genus-specific features. Analysis of specialized biosynthetic potential identified 13 gene clusters in MP11MiT and 4 in MP213FoT, highlighting their capacity for specialized metabolite biosynthesis. Based on phylogenomic, chemotaxonomic and phenotypic characteristics, strains MP11MiT and MP213FoT are proposed as representatives of two novel species, for which the names Gordonia abscondita sp. nov. (type strain MP11MiT=DSM 113667=CCM 9219) and Pseudochrobactrum mosebachae sp. nov. (type strain MP213FoT=DSM 113682=CCM 9218) are introduced.

Abstract This systematic study investigated the presence of total and third-generation cephalosporin-resistant (3GCR) Escherichia coli as well as total and benzyldimethyldodecylammonium chloride (BAC-C12) tolerant heterotrophic bacteria growing under nutrient-rich conditions at 37 °C, among them potential pathogens, in irrigation water (river and well water) potentially contaminated with wastewater, irrigated soils, and pepper (Capsicum annuum) fruits. Samples were collected from five vegetable-cultivating fields in Kosovo. The presence of E. coli was confirmed in both investigated rivers, all five examined wells, and in a single soil sample. Furthermore, 3GCR E. coli were detected in the two rivers and one well. Genomic fingerprinting revealed a high genetic variability among the cultivated E. coli strains. The 3GCR E. coli strains contained either bla CTX−M and bla TEM extended spectrum beta lactamase (ESBL) genes and/or exhibited an aminopenicillin-inactivating cephalosporinase (AmpC) phenotype. In addition, certain strains displayed resistance to ciprofloxacin and colistin, although none demonstrated resistance to carbapenems. Beyond (3GCR) E. coli, BAC-C12-tolerant bacteria were detected in river and well water, agricultural soil, and pepper fruit samples across all five investigated sites; among them bacteria of the genera Providencia, Morganella, and Pseudomonas. River water as a source of irrigation showed higher levels of microbial contamination compared to well water; however, well water also demonstrated contamination with fecal indicator bacteria. Despite this, no contamination of pepper fruits with fecal indicator bacteria was observed. Nonetheless, the presence of Pseudomonas with a high BAC-C12 tolerance at or in C. annuum fruits requires a more detailed examination.

Nature has been considered an interesting source of secondary bioactive compounds. Plants and their associated endophytes are common sources for these active constituents. Our study demonstrates the metabolomics profiling of the ethyl acetate extracts of three endophytic fungi associated with rosemary roots (Cladosporium spp., Alternaria spp. and Talaromyces spp.) in addition to the in vitro evaluation of the antitrypanosomal potential. The results revealed the presence of 47 metabolites from different chemical classes such as terpenes, phenolics, alkaloids, polyketides, macrolides, and others. Furthermore, the extracts of Cladosporium, Alternaria and Talaromyces exhibited potential inhibitory effects against T. brucei with IC50 values of 1.3, 3.2 and 3.5 µg/mL, respectively. Supporting the study, the identified compounds were docked against two proteins: Rhodesain in complex with a macrolactam inhibitor and ornithine decarboxylase in complex with a c-terminal fragment of antizyme. The docking simulations showed that most of the identified compounds have moderate to comparable docking score (S = - 3.82 to - 6.10 kcal/mol) within rhodesain active site. In addition, they showed weak to moderate docking scores (- 2.33 to - 5.9 kcal/mol) with a differential docking profile within ornithine decarboxylase active site. According to these findings, fungal endophytes associated with rosemary roots can be considered as a promising source of antitrypanosomal bioactive metabolites.

Two motile bacterial strains, designated as cfHf56-1 T and SW 252, were isolated from the coelomic fluid of Holothuria forskali and from the surrounding seawater at the animal sampling site, respectively. The sea cucumber was collected in the Glénan archipelago (Brittany, France). Strains cfHf56-1 T and SW 252 were Gram-stain-negative, non-spore-forming and rod-shaped bacteria. Colonies made on marine agar plates were brown in color. The pH and temperature ranges for growth were 7–8 and 18–30 °C, respectively, in marine broth. The major fatty acids were 16 : 1 cis9 and 16 : 0. Phylogenetic analyses evidenced that both strains belong to a novel species in the genus Pseudoalteromonas . The strains were closely related to the type strains of Pseudoalteromonas caenipelagi , Pseudoalteromonas byunsanensis and Pseudoalteromonas amylolytica, with 75–78 % ANI and 19–21 % dDDH values. In this context, cfHf56-1 T (= CIP 111854 T = CECT 30642 T ) is considered as the type strain of the novel species for which the name Pseudoalteromonas holothuriae sp. nov. is proposed. The genome of the type strain is characterized by a size of 5.1 Mbp and a G+C content of 40.5%.

Bacteria belonging to the genera Chryseobacterium, Sphingomonas, Variovorax, Neorhizobium, Parapedobacter and Erwinia have been isolated from different soils and have also been shown to exhibit plant growth promoting (PGP) features. A polyphasic approach was employed to ascertain the taxonomic status of eight selected strains all isolated from dry soils in association with plants showing high desiccation tolerances. Based on the result of the polyphasic characterization, the following names are proposed: Chryseobacterium hilariae sp. nov., with DT-3T as the type strain (= CIP 112171T = LMG 32722T = CCM 9258T); Sphingomonas astragali sp. nov., with DT-204T as the type strain (= CCM 9255T = LMG 33194T = CIP 112175T); Sphingomonas radicis sp. nov., with DT-207T as the type strain (= CCM 9257T = LMG 32727T = DSM 114514T); Sphingomonas larreae sp. nov., with DT-51T as the type strain (= CCM 9259T = CIP 112177T = DSM 114511T = LMG 32723T); Variovorax stachyos sp. nov., with DT-64T as the type strain (= LMG 32724T = DSM 114531T = CIP 112170T); Erwinia artemisiae sp. nov., with DT-104T as the type strain (CCM 9256T = LMG 32725T = DSM 114512T); Parapedobacter brassicae sp. nov., with DT-150T as the type strain (LMG 32759T = DSM 115120T); and Neorhizobium descurainiae sp. nov., with DT-125T as the type strain (LMG 32760T = CIP 112184T).

Members of the genera Flavobacterium, Chryseobacterium and Sphingomonas constitute a group of microorganisms in the rhizosphere associated with plant growth promoting (PGP) features. A polyphasic approach was employed to ascertain the taxonomic status of five selected strains. Overall genome relatedness indices of digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) between the strains and the other members of the genera Flavobacterium, Chryseobacterium and Sphingomonas were found to be below the established thresholds, respectively. Morphological, physiological, and biochemical characteristics of the strains confirmed their status as five novel species. A large variety of genes involved in plant growth promotion and carbohydrate utilization were found in all strains suggesting a contribution of all strains to PGP. Based on the result of the polyphasic characterization, the following names are proposed: Chryseobacterium terrae sp. nov., with the strain ST-37T as the type strain (= CCM 9260T = LMG 32728T); Flavobacterium plantiphilum sp. nov., with the strain ST-87T as the type strain CIP 112180T = DSM 114790T = LMG 32757T); Flavobacterium rhizophilum sp. nov., with the strain ST-75T as the type strain (= CIP 112185T = DSM 114831T = LMG 32758T); Flavobacterium rhizosphaerae sp. nov., with the strain ST-119T as the type strain (CIP 112181T = DSM 114832T = LMG 32756T); and Sphingomonas plantiphila sp. nov. with the strain ST-64 T as the type strain (= CCM 9261T = CIP 112178T = DSM 114515T = LMG 32729T).

Methicillin-resistant Staphylococcus aureus (MRSA) is a significant pathogen associated with healthcare-related infections that are often challenging to treat. Conditions such as, skin and soft tissue infections, bloodstream infections, and pneumonia highlight the critical need for effective therapeutic strategies. Careful use of antibiotics under medical supervision is essential to prevent the further emergence of MRSA. Recent studies have documented the antibacterial efficacy of certain endophytic fungi extracts against MRSA, suggesting their potential as a source of novel treatments. This study investigates the metabolomic profiling of the endophytic fungus Aspergillus sp. SH1 using liquid chromatography-high-resolution electrospray ionization mass spectrometry (LC-HR-ESI-MS) and evaluates the anti-MRSA potential of the fungal extract. The metabolomic analysis identified 27 compounds (1–27) with diverse chemical natures, including polyketides, alkaloids, cyclic tripeptides, polypropionate derivatives, and sesquiterpenes. The fungal extract exhibited potent anti-MRSA activity, with an IC50 value of 9.8 µg/mL, compared to ciprofloxacin (IC50 = 25.7 µg/mL). To support these findings, in silico studies were performed to model the binding interactions of the identified compounds with key MRSA-related targets, including Toll-like receptor 2 (TLR2), von Willebrand factor (VWF), tumor necrosis factor (TNF), and penicillin-binding protein 2a (PBP2a). Compounds 2, 9, 15, 16, 20, 22, and 25 demonstrated enhanced binding affinities, suggesting their potential as lead molecules for developing new antibacterial agents targeting MRSA. In conclusion, this study highlights the promising anti-MRSA potential of Aspergillus sp. SH1 extract, providing a foundation for further exploration of its bioactive compounds in combating resistant bacterial infections.

High-rate algae ponds (HRAPs) efficiently remove nitrogen and phosphorus from wastewater and the produced microalgal biomass can be used as NPK-rich fertilizer in a sanitation‒agricultural circular economy. The effects of microalgal based organic fertilizer on plant growth are well documented, but the knowledge of effects on the plant microbiome is scarce. We analysed the rhizosphere and root microbiome of wheat seven weeks after fertilization with microalgal biomass harvested from a HRAP used for municipal wastewater treatment. Plants fertilized with a NPK mineral fertilizer, unfertilized control plants, and the microalgal biomass were studied in parallel. Three fertilizer levels, 60, 120, and 180 kg N ha-1, were compared. The bacterial community associated with microalgal biomass was dominated by few taxa including potential plant growth-promoting bacteria (PGPBs) and potentially pathogenic bacteria. Root-associated bacterial communities were more sensitive to fertilization than those of the rhizosphere. Fertilization with microalgal biomass increased the bacterial abundance in roots, but the diversity and composition of the root microbiome hardly changed. Only few taxa increased significantly in relative abundance after fertilization with microalgal biomass, while the mineral fertilizer reduced the bacterial abundance and diversity in roots. Escherichia coli, 3rd generation cephalosporine resistant (3GCR) E. coli, and enterococci were cultivated only from microalgal biomass and plants fertilized with it, while potentially pathogenic bacteria tolerant to biocidal compounds and antibiotic resistance genes (ARGs) were detected in all samples. Although the microalgae-based organic fertilizer derived from a wastewater treating HRAP provides a sustainable source of nutrients, its contribution to the spread of antibiotics, biocidal compounds, and antimicrobial resistance in agricultural systems should be further evaluated.

Cancer incidence continues to increase every year. Scientists strive to search for new anticancer compounds to combat this disease. Appealingly, marine environmental niches are still an untapped scaffold for natural products with chemical and biomedical diversity. Hence, fungi isolated from the Red Sea sponge Spheciospongia vagabunda were explored. Two strains were purified from the sponge and identified, depending on 18 S rRNA gene sequence, as Aspergillus sp. (UR1) and Penicillium sp. (UR2). The obtained fungal extracts were submitted to LC-HR-ESI-MS metabolomics evaluation, which showed notable variation in the chemical profiles of both extracts. The cytotoxic activity was assessed against three cancer cell lines: HepG2 (hepatocellular carcinoma human), MCF7 (breast cancer) and CaCo-2 (human colon carcinoma), via MTT assay. UR1 extract displayed higher antiproliferative activity with IC 50 values 2.61 ± 0.12, 3.23 ± 0.21 and 3.41 ± 0.18 µg/ml against HepG2, CaCo-2 and MCF7, respectively. Whereas UR2 extract exhibited a less potent effect with IC 50 values of 17.65 ± 0.28, 18.38 ± 0.19, and 22.45 ± 0.27 µg/ml. Additionally, molecular docking was conducted. Most identified compounds established strong binding affinity with the PPARG gene. Compounds 6 and 16 showed binding energy with S values − 9.13 and − 8.38 kcal/mol, respectively. The findings suggested the importance of Spheciospongia vagabunda -derived fungi in the production of cytotoxic natural compounds that could be used for cancer management.

Abstract Background Sponge-associated microorganisms are promising resources for the production of bioactive compounds with cytotoxic potential. The main goal of our study is to isolate the fungal endophytes from the Red Sea sponge Hyrtios sp. followed by investigating their cytotoxicity against number of cell lines. Results The fungal strain UR3 was isolated from the Red Sea sponge using Sabouraud dextrose agar media. It was identified based on partial 18 S rRNA gene and ITS sequence analyses as Cladosporium sp. UR3. The in vitro cytotoxic potential of the ethyl acetate extract of the fungal isolate was evaluated using MTT assay against three cancer cell lines: CACO2, MCF7, and HEPG2. Metabolomics profiling of the obtained ethyl acetate extract using LC-HR-ESI-MS, along with molecular docking and pharmacological network studies for the dereplicated compounds were performed to explore its chemical profile and the possible cytotoxic mechanism of the sponge-associated fungi. Conclusion These results highlighted the role of sponge-associated fungi as a fruitful resource for the discovery of cytotoxic metabolites.

A Gram-staining-positive, aerobic bacterium, designated strain JJ-181 T, was isolated from the root surface of soybean. Based on the 16S rRNA gene sequence similarities, strain JJ-181 T was grouped into the genus Cohnella, most closely related to Cohnella hashimotonis F6_2S_P_1T (98.85

Endophytic fungi are known to be a rich source for anti-infective drugs. In this study, Aptenia cordifolia associated endophytic fungi were explored for the first time. Seven isolates were identified morphologically followed by screening of these fungi by plug diffusion assay which revealed their potential activity against Staphylococcus aureus (ATCC 9144), Bacillus cereus (ATCC 14579), Serratia marcescens (ATCC 14756), Fusarium oxysporum (ATCC 48112), and Aspergillus flavus (ATCC 22546). Additionally, the crude ethyl acetate extract of the most potent three isolates in plug diffusion assay showed that Aspergillus sp. ACEFR2 was the most potent as anti-infective in disc diffusion assay; Accordingly, Aspergillus sp. ACEFR2 was investigated using phylogenetic analysis and LC-HR-ESI-MS. The phylogenetic analysis placed the strain into the Aspergillus section Niger close related to few species including A. niger. Whereas the metabolomic profiling revealed the presence of diverse pool of metabolites. Furthermore, in silico molecular docking study was carried out to predict which compounds most likely responsible for the anti-infective activity.

Twenty fungal endophytes were isolated from Nabq, and plug diffusion screening revealed four of those isolates to be promising. These isolates were identified molecularly as Aspergillus sp. of Sect. Flavi (ZSEFL2) and Sec. Niger (ZSEFL14), and Penicillium spp. closest related to Penicillum crustosum (ZSEFL15, 19). Investigations were conducted to determine the effects of culture parameters, such as the type of culture medium, incubation temperature, initial pH and agitation, on the production of antimicrobial metabolites in a culture filtrate extract. Using the disk diffusion technique, antimicrobial metabolite synthesis was evaluated. In comparison to the other three distinct culture media (PDB-y, rice, and Cz), PDB demonstrated to be the best culture medium for the four tested fungal endophytes, exhibiting high levels of the Antimicrobial Index (AI). Also, a restricted range of incubation temperatures resulted in the formation of bioactive metabolites, but raising the incubation temperatures from 25 to 30 degrees C increased the amount of bioactive metabolites produced. Moreover, the best initial hydrogen concentration typically ranged from pH 5 to 6. Finally, when the conditions changed from stationary to shaking, the bioactive secondary metabolite's antimicrobial efficacy reduced. The behavior of the four examined fungal isolates was consistent, and they all required the same conditions to produce the most bioactive metabolites. Moreover, ethyl acetate extract's MIC values against the investigated bacterial and fungal human pathogens ranged from 0.19 to 6.25 mg/mL. (c) 2023 SAAB. Published by Elsevier B.V. All rights reserved.

The genus Vagococcus belongs to the family Enterococcaceae (order Lactobacillales) and is closely related to the genus Enterococcus. Currently, 19 species of the genus have been validly named. In this study, we isolated strain G314FT from the common green bottle fly Lucilia sericata collected in Germany. Sequencing of its almost- complete 16S rRNA gene revealed that the isolate belongs to the genus Vagococcus, being closely related to Vagococcus bubulae SS1994T with high sequence identity (99.50 %), followed by Vagococcus martis D7T301T (98.86 %), Vagococcus vulneris SS1995T (98.71 %), Vagococcus teuberi DSM 21459T (98.64 %), Vagococcus silagei 2B-2T (98.64 %) and Vagococcus penaei CD276T (98.64 %). Genome sequencing of strain G314FT was performed by a combination of Illumina and Oxford Nanopore technology, yielding a circular genome with a size of 2 139 468 bp and an 11 kb plasmid. Average nucleotide identity and digital DNA-DNA hybridization values were calculated between G314FT and its closest- related taxa, and found to be <91 % and <40 %, respectively, thus strongly supporting that strain G314FT represents a novel species of the genus Vagococcus. Phylogenetic and core protein- based phylogenomic trees revealed that G314FT was closely related to a group of three species, V. bubulae SS1994T, V. martis D7T301T and V. teuberi DSM 21459T. Comparatively, the genome of G314FT is the smallest in the group of the four related species, and the biochemical pathway comparison using BlastKOALA revealed that G314FT has lost some amino acid biosynthetic proteins; however, it has gained enzymes for carbohydrate metabolism. Phenotypically, G314FT was consistent with other species of the genus Vagococcus including a negative catalase reaction and non- motility. Using the polyphasic approach, our data supports that the isolate represents a new species, for which we propose the name Vagococcus luciliae G314FT (=DSM 112651T= CCM 9164T).

A Gram-strain positive, aerobic, endospore-forming bacterial strain (JJ-246T) was isolated from the rhizosphere of Zea mays. The 16S rRNA gene sequence similarity comparisons showed a most closely relationship to Paenibacillus oenotherae DT7-4T (98.4%) and Paenibacillus xanthinolyticus 11N27T (98.0%). The pairwise average nucleotide identity and digital DNA-DNA hybridisation values of the JJ-246T genome assembly against publicly available Paenibacillus type strain genomes were below 82% and 33%, respectively. The draft genome of JJ-246T shared many putative plant-beneficial functions contributing (PBFC) genes, related to plant root colonisation, oxidative stress protection, degradation of aromatic compounds, plant growth-promoting traits, disease resistance, drug and heavy metal resistance, and nutrient acquisition. The quinone system of strain JJ-246T, the polar lipid profile and the major fatty acids were congruent with those reported for members of the genus Paenibacillus. JJ-246T was shown to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus plantiphilus sp. nov. is proposed, with JJ-246T (= LMG 32093T = CCM 9089T = CIP 111893T) as the type strain.

Endophytic fungi are known to be a rich source of anti-infective drugs. In our study, Allium cepa was investigated for fungal diversity using different media to give 11 isolates which were identified morphologically. Out of the isolated fungal strains, Penicillium sp. (LCEF10) revealed potential anti-infective activity against the tested microbes (Fusarium solani ATTC 25922, Pseudomonas aeruginosa (ATTC 29231), Staphylococcus aureus ATTC 27853, Candida albicans ATTC 10231), besides, their MICs were measured by well diffusion method, therefore, it was subjected to molecular identification in addition to phylogenetic analysis. Moreover, the ITS sequence of strain LCEF10 showed a consistent assignment with the highest sequence similarity (99.81%) to Penicillium oxalicum NRRL 787. The crude ethyl acetate extract of Penicillium sp. LCEF10 was investigated for metabolomic analysis using LC-HR-ESI-MS. The metabolic profiling revealed the presence of polyketides, macrolides, phenolics and terpenoids. Furthermore, in silico molecular docking study was carried out to predict which compounds most likely responsible for the anti-infective activity.

Endophytic fungi, particularly from higher plants have proven to be a rich source of antimicrobial secondary metabolites. The purpose of this study is to examine the antimicrobial potential of three endophytic fungi Aspergillus sp. SA1, Aspergillus sp. SA2, and Aspergillus sp. SA3, cultivated from Nigella sativa seeds against Staphylococcus aureus (ATCC 9144), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), Klebsiella pneumoniae (ATCC 13883), MRSA (ATCC 33591), and human pathogen Candida albicans (ATCC 10231). Furthermore, the most active cultivated endophytic fungi were molecularly identified via internal transcribed spacer (ITS) sequencing. HR-ESIMS guided approach has been used successfully in chemical profiling of 26 known bioactive secondary metabolites (1-26), which belongs to different classes of natural compounds such as polyketides, benzenoids, quinones, alcohols, phenols or alkaloids. Finally, in-silico interactions within active site of fungal Cyp51 and bacterial DNA gyrase revealed possibility of being a hit-target for such metabolites as antimicrobials.