Serratia marcescens are considered to be abundant and optimal resources for obtaining prodigiosin, which can be isolated from soil, water, plants and air but rarely from insects. In the present study, a strain of Serratia marcescens named WA12-1-18 was isolated from the gut of Periplaneta americana, which was capable of producing high levels of pigment reaching 2.77 g/l via solid fermentation and was identified as prodigiosin by ultraviolet, high performance liquid chromatography (LC), Fourier-transform infrared spectroscopy, LC-mass spectroscopy and nuclear magnetic resonance. The apoptotic tumor cells treated with prodigiosin were examined by 4',6-diamidino-2-phenylindole (DAPI) staining assays and transmission electron microscopy. Flow cytometry (FCM) was utilized to measure the apoptotic rate with Annexin V staining and the expression levels of proteins involved in apoptosis, including B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X (Bax) and caspase-3 were determined by western blot analysis and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The experimental results revealed that prodigiosin could inhibit the proliferation of HeLa cells and the half-maximal inhibitory concentration values of prodigiosin in HeLa were 2.1, 1.2 and 0.5 µg/ml over 24, 48 and 72 h, respectively. Furthermore, DAPI staining assays and transmission electron microscopy clearly demonstrated that prodigiosin could induce HeLa cell apoptosis. FCM results revealed that the cell apoptotic rates were 19.7±1.4, 23.7±2.4 and 26.2±2.3% following the treatment with 0.5, 1.0 and 2.0 µg/ml prodigiosin for 48 h, respectively. Western blot analysis and RT-qPCR revealed that prodigiosin could activate apoptosis-associated molecules including Bcl-2, Bax and caspase-3. Therefore, the results of the present study demonstrated that the prodigiosin could induce apoptosis in HeLa cells, which may be associated with the upregulation of Bax and caspase-3, the concomitant downregulation of Bcl-2 levels and also triggering the extrinsic apoptotic signaling pathway.

Methicillin-resistant Staphylococcus aureus (MRSA) is recognized as one of the serious pathogen that causes acquired infections worldwide. Its emerging need to discover novel, safe and potent anti-MRSA drugs. In this study, primary screening by anti-MRSA activity assay found one strain WA5-2-37 isolated from the intestinal tract of Periplaneta americana, exhibited great activity against MRSA ATCC 43300. The strain WA5-2-37 produced actinomycin X 2 and collismycin A which showed strong inhibition of MRSA with minimum inhibitory concentration (MIC) values of 0.25 and 8 µg/mL. The structures of the pure compounds were elucidated by analysis of mass spectrometry (MS), 1 H and 13 C nuclear magnetic resonance (NMR). The strain WA5-2-37 was considered as Streptomyces globisporus on the basis of morphological characteristics, genotypic data, and phylogenetic analysis. This is the first reported naturally occurring strain of S. globisporus isolated from the intestinal tract of P. americana, whereas it has almost been found from plants, marine, and soil previously. Moreover, S. globisporus has not been reported to produce any anti-MRSA substances previously, such as actinomycin X 2 and collismycin A. In conclusion, the insect-derived strain of S. globisporus WA5-2-37 was considered of great potential as a new strain of producing actinomycin X 2 , collismycin A or other anti-MRSA compounds.

IntroductionVibrio parahaemolyticus is a foodborne pathogen commonly found in seafood, and drug resistance poses significant challenges to its control. This study aimed to identify novel drug targets for antibacterial drug discovery.MethodsTo identify drug targets, we performed a pan-genome analysis on 58 strains of V. parahaemolyticus genomes to obtain core genes. Subsequently, subtractive proteomics and physiochemical checks were conducted on the core proteins to identify potential therapeutic targets. Molecular docking was then employed to screen for anti-V. parahaemolyticus compounds using a in-house compound library of Streptomyces parvus, chosen based on binding energy. The anti-V. parahaemolyticus efficacy of the identified compounds was further validated through a series of experimental tests.Results and DiscussionPangenome analysis of 58 V. parahaemolyticus genomes revealed that there were 1,392 core genes. After Subtractive proteomics and physiochemical checks, Flagellar motor switch protein FliN was selected as a therapeutic target against V. parahaemolyticus. FliN was modeled and docked with Streptomyces parvus source compounds, and Actinomycin D was identified as a potential anti-V. parahaemolyticus agent with a strong binding energy. Experimental verification confirmed its effectiveness in killing V. parahaemolyticus and significantly inhibiting biofilm formation and motility. This study is the first to use pan-genome and subtractive proteomics to identify new antimicrobial targets for V. parahaemolyticus and to identify the anti-V. parahaemolyticus effect of Actinomycin D. These findings suggest potential avenues for the development of new antibacterial drugs to control V. parahaemolyticus infections.

Gut microbes play critical roles in host nutrition, physiology, and behavior. Periplaneta americana is a famous urban pest which is widely distributed in the tropics and subtropics, but very few information is available on the gut microbiome of Periplaneta americana, particularly in its different life stages. Here, we characterized the diversity and structure of gut microbiome in eggs, nymph and adult life stages of Periplaneta americana using high-throughput 16S rRNA genes sequencing. Both the results of Alpha- and Beta-diversity analysis showed the diversity and structure of gut microbiome were significant different among the eggs, nymph and adult stages. The result of species distribution showed the predominant phyla in three life stages were Bacteroidetes , Firmicutes and Proteobacteria , but the relative abundances of these bacteria were significant different among each life stage. 1,169 operational taxonomic units were shared by three stages, which indicating the gut microbiome may be inherited to offspring from parents of Periplaneta americana. According to the prediction of functional genes in metabolic pathways, most of them were distributed in the metabolic pathways of basic physiology such as nutrition, growth, development and immunity, etc. The relative abundances of functional genes in metabolic pathways were significant different among life stages of Periplaneta americana, indicating the gut microbiome might play an important role in the physiology across its different life stages. This study revealed the diversity and structure of gut microbiome in different life stages of Periplaneta americana, which may contribute to us to understand it’s physiology and behaviors.