The effect of stimulating Lucilia sericata larvae with Staphylococcus aureus on the production of bioactive components in larval secretions - dataset.

The dataset comprises: i) Chromatograms of induced and uninduced <3 kDa L. sericata larval secretions (Figures 1-4). Chromatographic separation of induced and uninduced Ls larval S/E fractions (<3 kDa) was performed on a Superdex 30 Increase 10/300 GL column. Samples were eluted isocratically with deionised water (0.5 mL/min), and elution was monitored at 215 nm ii) Evaluation of larval secretion effects on HDMEC and HUVEC cell proliferation using the MTS assay Cells (Human Dermal Microvascular Endothelial Cells, HDMEC oraz Human Umbilical Vein Endothelial Cells, HUVEC) were seeded at 4×10³ cells per well in 150 µL complete medium. After 24 h, medium was replaced with serum- and supplement-free medium containing larval secretion fractions. Following 48 h incubation, cell proliferation was assessed using the MTS assay. Briefly, 20 µL MTS reagent and 100 µL fresh medium were added per well and incubated for 2 h at 37°C. Absorbance was measured at 490 nm using a microplate reader, and viability was expressed relative to untreated controls (100%). The results of the absorbance measurements in the MTS assay, as presented in the files (MTS_absorbance_DERMAL_raw data + descriptions, MTS_absorbance_HUVEC _raw data + descriptions) Figure 5 Proliferation of HDMECs and HUVECs following 48 h treatment with induced LMW secretion fractions The Y-axis shows absorbance (a.u.) at 490 nm, reflecting mitochondrial metabolic activity and indicating cell proliferation. The darker bars represent cells within the treatment condition (receiving volumetric doses of WF fractions), while the lighter bars represent the untreated control baseline. Asterisks (*) indicate P<0.05 based on Tukey's HSD with Bonferroni correction. iii) Effect of induced fractions on bacterial growth (Radial diffusion assay) - photo An overnight culture of the reference bacteria was diluted 1:10 and adjusted to a turbidity of 0.5 McFarland in MH broth. This bacterial suspension was then incorporated into molten MH II agarose (0.75% w/v) before being poured onto sterile Petri dishes. Once solidified, 20 µL of each sample was applied to the surface of the agarose. The plates were then incubated overnight at 32 °C and the inhibition zones evaluated the following day. The results were presented at: Figure 6. Assessment fractions activity against Pseudomonas aeruginosa via radial diffusion assay Selected individual induced fractions. The distinct zones of depigmentation (loss of pyocyanin) against P. aeruginosa are clearly visible for fractions WF1 and WF4 (indicated by a red arrow), distinguishing them from the surrounding bacterial lawn. iv) Effect of induced fractions on bacterial growth (Microtiter cell viability assay) The antimicrobial activity of fractions was assessed against the target bacterial strain using a PrestoBlue-based microtiter viability assay. Briefly, 20 µL of sample, 80 µL of MH broth, and 5 µL of bacterial inoculum were combined in 96-well plates. Untreated cultures and broth-only wells served as controls. Plates were incubated at 32 °C with shaking (280–300 rpm) for 16–20 h. Growth inhibition was evaluated visually, followed by the addition of PrestoBlue reagent and a further 3 h incubation. Fluorescence was measured at 560/590 nm (excitation/emission). The results of the fluorescence measurements in the PrestoBlue assay, as presented in the file: PrestoBlue_fluorescence_raw data + descriptions. Figure 7. Metabolic activity of P. aeruginosa and S. aureus following exposure to induced fractions (mean ± confidence interval) The Y-axis represents the Relative Fluorescence Units (RFU) measured at 560/590 nm (excitation/emission). Asterisk (*) indicates a significant difference in comparison to control (Tukey pairwise comparison and with Bonferroni correction, P<0.05). The collected material was used to prepare the publication: DOI: 10.32383/appdr/219760 The work is supported by the National Science Centre, Poland: UMO-2021/43/D/NZ7/03119.