Antibiotics, nowadays, are not only used for the treatment of human diseases but also used in animal and poultry farming to increase production. Overuse of antibiotics leads to their circulation in the food chain due to unmanaged discharge. These circulating antibiotics and their residues are a major cause of antimicrobial resistance (AMR), so comprehensive and multifaceted measures aligning with the One Health approach are crucial to curb the emergence and dissemination of antibiotic resistance through the food chain. Different chromatographic techniques and capillary electrophoresis (CE) are being widely used for the separation and detection of antibiotics and their residues from food samples. However, the matrix present in food samples interferes with the proper detection of the antibiotics, which are present in trace concentrations. This review is focused on the scientific literature published in the last decade devoted to the detection of antibiotics in food products. Various extraction methods are employed for the enrichment of antibiotics from a wide variety of food samples; however, solid-phase extraction (SPE) techniques are often used for the extraction of antibiotics from food products and biological samples. In addition, this review has scrutinized how changing instrumental composition, organization, and working parameters in the chromatography and CE can greatly impact the identification and quantification of antibiotic residues. This review also summarized recent advancements in other detection methods such as immunological assays, surface-enhanced Raman spectroscopy (SERS)-based assays, and biosensors which have emerged as rapid, sensitive, and selective tools for accurate detection and quantification of traces of antibiotics.

Finding new antibacterial agents from natural products is urgently necessary to address the growing cases of antibiotic-resistant pathogens. Actinomycetes are regarded as an excellent source of therapeutically important secondary metabolites including antibiotics. However, they have not yet been characterized and explored in great detail for their utility in developing countries such as Nepal. In silico molecular docking in addition to antimicrobial assays have been used to examine the efficacy of chemical scaffolds biosynthesized by actinomycetes. This paper depicts the characterization of actinomycetes based on their morphology, biochemical tests, and partial molecular sequencing. Furthermore, antimicrobial assays and mass spectrometry-based metabolic profiling of isolates were studied. Seventeen actinomycete-like colonies were isolated from ten soil samples, of which three isolates showed significant antimicrobial activities. Those isolates were subsequently identified to be Streptomyces species by partial 16S rRNA gene sequencing. The most potent Streptomyces species_SB10 has exhibited an MIC and MBC of 1.22 μg/mL and 2.44 μg/mL, respectively, against each Staphylococcus aureus and Shigella sonnei. The extract of S. species_SB10 showed the presence of important metabolites such as albumycin. Ten annotated bioactive metabolites (essramycin, maculosin, brevianamide F, cyclo (L-Phe-L-Ala), cyclo (L-Val-L-Phe), cyclo (L-Leu-L-Pro), cyclo (D-Ala-L-Pro), N6, N6-dimethyladenosine, albumycin, and cyclo (L-Tyr-L-Leu)) were molecularly docked against seven antimicrobial target proteins. Studies on binding energy, docking viability, and protein-ligand molecular interactions showed that those metabolites are responsible for conferring antimicrobial properties. These findings indicate that continuous research on the isolation of the Streptomyces species from Nepal could lead to the discovery of novel and therapeutically relevant antimicrobial agents in the future.

Background/Objectives: Recently, antimicrobial-resistant pathogens and cancers have emerged as serious global health problems, highlighting the immediate need for novel therapeutics. Consequently, we aimed to isolate and characterize endophytic Streptomyces strains from the rhizospheres of the Himalayan region of Nepal and identify specialized metabolites with antibacterial, antifungal, and cytotoxic potential. Methods: To isolate Streptomyces sp., we collected two soil samples and cultured them on an ISP4 medium after pretreatment. We isolated and identified the strains PY108 and PY109 using a combination of morphological observations and 16S rRNA gene sequencing. Results: The BLAST results showed that PY108 and PY109 resembled Streptomyces hundungensis PSB170 and Streptomyces sp. Ed-065 with 99.28% and 99.36% nucleotide similarity, respectively. Antibacterial assays of ethyl acetate (EA) extracts from both isolates PY108 and PY109 in a tryptic soy broth (TSB) medium were conducted against four pathogenic bacteria. They showed significant antibacterial potential against Staphylococcus aureus and Klebsiella pneumoniae. Similarly, these extracts exhibited moderate antifungal activities against Saccharomyces cerevisiae and Aspergillus niger. Cytotoxicity assays on cervical cancer cells (HeLa) and breast cancer cells (MCF-7) revealed significant potential for both extracts. LC-MS/MS profiling of the EA extracts identified 27 specialized metabolites, including diketopiperazine derivatives, aureolic acid derivatives such as chromomycin A, and lipopeptide derivatives. In comparison, GC-MS analysis detected 34 metabolites, including actinomycin D and γ-sitosterol. Furthermore, a global natural product social molecular networking (GNPS)-based molecular networking analysis dereplicated 24 metabolites in both extracts. Conclusions: These findings underscore the potential of endophytic Streptomyces sp. PY108 and PY109 to develop new therapeutics in the future.

Actinomycetes are a unique class of Gram-positive bacteria with a fungus-like filamentous mycelium. They are often called ‘antibiotics factories’ because they can produce many potent secondary metabolites. At present, multidrug-resistant (MDR) pathogens are the main problem for humans and animals and to overcome this problem, more potent antibiotics are urgently required. Fortunately, Nepal has great geographical and climatic diversity serving as a home for various actinomycetes. This study identified several actinomycetes using morphological and biochemical assays from seven soil samples collected from different elevations in Nepal. The primary objective of this study was to screen the potent metabolites-producing actinomycetes. For this purpose, antimicrobial assays of seven isolates were carried out against Escherichia coli ATCC 25922, Salmonella typhi ATCC 14028, Shigella sonnei ATCC 25931, and Staphylococcus aureus ATCC 43300. The sample BT36 demonstrated the highest zone of inhibition against the tested microorganisms among others. Similarly, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of all isolates were carried out. Among them, the MIC of samples BT36, PT7, and AB1 were found to be 1.952 mg/mL, 0.012 mg/mL, and 0.029 mg/mL, respectively. Ethyl acetate extracts of resulting potent isolates PT7 and AB1 were subjected to mass spectrometric analysis. A total of 22 bioactive compounds were annotated from liquid chromatography-high resolution mass spectroscopy (LC-HRMS), including epopromycin A, myxopyronin B, gilvocarcin HE, and okilactomycin A. It is concluded that this study will be useful in setting up a strategy to isolate useful secondary metabolites from soil microbes from the hilly region of Nepal.

Metabolic profiling of actinomycetes isolated from Nepalese soil has uncovered promising candidates for the development of new antimicrobial and anticancer agents. This research examines the potential of actinobacteria isolated from soil, namely strains PY40, PY118, and PY122, as promising sources for biological applications. These strains were identified using 16S rRNA gene sequencing as Streptomyces sp. PY40, Streptomyces sp. PY118, and Amycolatopsis sp. PY122. Comprehensive bioassays were carried out on ethyl acetate (EA) extract originating from the fermentation broths of these species to assess their antimicrobial and anticancer activities. The results showed noteworthy antifungal and anticancer properties of Streptomyces sp. PY40 extract, thereby indicating its possible use in the synthesis of pharmaceuticals. Furthermore, potent antifungal activity was observed with an extract from Amycolatopsis sp. PY122. These isolates showed strong inhibition against several multi-drug-resistant strains of bacteria, including Shigella sonnei ATCC 25931, Klebsiella pneumoniae ATCC 700603, Escherichia coli ATCC 25922, and, Staphylococcus aureus ATCC 43300, suggesting significant antibacterial activity of the extracts. Gas Choursomatography-Mass Spectrometry (GC-MS) analysis revealed particularly volatile chemical constituents in the extracts. Antibiotic agents such as diketopiperazines and hexahydropyrrolo[1,2-a]pyrazine-1,4-dione were prevalent in the extracts of these isolates. This study enhances our comprehension of the bioactive potential of these microorganisms and underscores their value as a source for creating novel therapeutics to address multi-resistant infections and cancer demands. These findings show that actinomycetes from Nepalese soil have high prospects for biological applications.