Entomopathogenic nematodes (EPN) species differ in their capability to withstand rapid desiccation (RD). Infective juveniles of Steinernema carpocapsae are a better adaptable and tolerant than Steinernema feltiae or Heterorhabditis bacteriophora as, an optimal RH of > 90% is required by S. feltiae and H. bacteriophora while maintaining RH equivalent to 74% could sustain survival of S. carpocapsae under RD. Our findings from infectivity suggest that following application, shrunk IJs are acquired passively by the larvae, probably rehydrate and resume infection within the insect gut. Water loss rate is a key factor affecting survival of S. carpocapsae on exposed surfaces. The present study provides the foundation for characterizing mechanism of rapid rate of water loss in EPN. ATR-FTIR is a rapid and reliable method for analysis of water loss. Changes in peak intensity was observed at 3100–3600 cm−1 (OH bonds of water), 2854 cm−1 (CH stretching of symmetric CH2, acyl chains), 2924 cm−1 (CH stretching of anti-symmetric CH2, lipid packing heterogeneity), 1634 cm−1 (amide I bonds) indicate major regions for hydration dependent changes in all EPN species. FTIR data also indicates that, S. carpocapsae contains strong water interacting regions in their biochemical profile, which could be an influencing factor in their water holding capacity under RD. ATR-FTIR were correlated to water content determined gravimetrically by using Partial Least square –Regression and FTIR multivariate method, which could be used to screen a formulation’s potential to maintain or delay the rate of water loss in a rapid and efficient manner.
This study presents a new eco-friendly formulation of entomopathogenic nematodes (EPNs) based on individual coating of EPNs with titanium dioxide (TiO 2 ) nanoparticles (NPs) and mineral oil via oil-in-water Pickering emulsions. Mineral oilin-water emulsions stabilized by amine-functionalized titanium dioxide (TiO 2 -NH 2 ) particles were prepared. 40:60 and 50:50 oil− water volume ratios using 2 wt % TiO 2 -NH 2 particles were found to be the most stable emulsions with a droplet size suitable for the formulation and were further studied for their toxicity against the incorporated EPNs. Carboxyfluorescein was covalently bonded to TiO 2 -NH 2 NPs, and the resulting composite was observed via fluorescence confocal microscopy. The dry coating was evaluated using SEM and confocal microscopy, which showed significant nematode coverage by the particles and oil. The final formulation was biocompatible with the studied EPNs, where the viability of the EPNs in the formulation was equivalent to control aqueous suspension after 120 days. Finally, yields of nematodes from infected Galleria mellonella cadavers collected for 150 days showed no significant differences (P > 0.05) using the tested emulsions compared to the control containing nematodes in water.
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