Top‐down population regulation can influence the success of biological control agents when they are released into the field. Entomopathogenic nematodes (EPNs) are used commonly in biological control programmes, but their efficacy suffers from poor persistence. Although abiotic soil conditions have been shown to reduce EPN persistence, consumption of infected insects by scavengers and of infective juvenile (IJ) nematodes by predators may also regulate these populations. In the present study, the effects of different soil arthropods on EPNs in laboratory conditions were measured. It was hypothesised that arthropods commonly found in soil communities where EPNs are applied would consume cadavers of insects parasitised by the nematodes and the IJs themselves. Some species of scavengers consume EPN‐infected insects. Crickets (Gryllus bimaculatus De Geer), American cockroaches [Periplaneta Americana (Linnaeus)], ants [Tetramorium chefketi Forel and Pheidole pallidula (Nylander)], earwigs (Labidura riparia Pallas), mites (Sancassania polyphyllae Zachvatkin), and springtails (Sinella curviseta Brook and Folsomia candida Willem) have different responses to nematode‐killed insects. Results suggested that ants (T. chefketi), cockroaches, mites, and earwigs fed on Steinernema‐killed insects whereas neither crickets nor springtails consumed them. In the second part of the study, experiments were conducted to determine whether mites and springtails consumed IJ EPNs. Results showed that S. polyphyllae mites do not consume infective juveniles in soil, whereas both springtail species consumed significant numbers of the IJs. Top‐down regulatory processes can be a limiting factor for EPN populations under laboratory conditions. Both host cadavers and IJs are consumed (albeit by different arthropods), so these results may help explain the difficulties associated with the persistence of EPN application to soil.

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Natural products have been proven to be important starting points for the development of new drugs. Bacteria in the genera Photorhabdus and Xenorhabdus produce antimicrobial compounds as secondary metabolites to compete with other organisms. Our study is the first comprehensive study screening the anti-protozoal activity of supernatants containing secondary metabolites produced by 5 Photorhabdus and 22 Xenorhabdus species against human parasitic protozoa, Acanthamoeba castellanii, Entamoeba histolytica, Trichomonas vaginalis, Leishmania tropica and Trypanosoma cruzi, and the identification of novel bioactive antiprotozoal compounds using the easyPACId approach (easy Promoter Activated Compound Identification) method. Though not in all species, both bacterial genera produce antiprotozoal compounds effective on human pathogenic protozoa. The promoter exchange mutants revealed that antiprotozoal bioactive compounds produced by Xenorhabdus bacteria were fabclavines, xenocoumacins, xenorhabdins and PAX peptides. Among the bacteria assessed, only P. namnaoensis appears to have acquired amoebicidal property which is effective on E. histolytica trophozoites. These discovered antiprotozoal compounds might serve as starting points for the development of alternative and novel pharmaceutical agents against human parasitic protozoa in the future.

Xenorhabdus and/or Photorhabdus bacteria produce antibacterial metabolites to protect insect cadavers against food competitors allowing them to survive in nature with their nematode host. The effects of culture supernatant produced by Xenorhabdus and Photorhabdus spp. were investigated against the multidrug-resistant dental root canal pathogen Enterococcus faecalis. The efficacy of seven different cell-free supernatants of Xenorhabdus and Photorhabdus species against E. faecalis was assessed with overlay bioassay and serial dilution techniques. Additionally, time-dependent inactivation of supernatant was evaluated. Among the seven different bacterial species, X. cabanillasii produced the strongest antibacterial effects. Loss of bioactivity in a phosphopantetheinyl transferase-deficient mutant of X. cabanillasii indicated that this activity is likely based on non-ribosomal peptide synthetases (NRPSs) or polyketide synthases (PKSs). Subsequent in silico analysis revealed multiple possible biosynthetic gene clusters (BGCs) in the genome of X. cabanillasii including a BGC homologous to that of zeamine/fabclavine biosynthesis. Fabclavines are NRPS-derived hexapeptides, which are connected by PKS-derived malonate units to an unusual polyamine, also PKS-derived. Due to the known broad-spectrum bioactivity of the fabclavines, we generated a promoter exchange mutant in front of the fabclavine-like BGC. This leads to over-expression by induction or a knock-out by non-induction which resulted in a bioactive and non-bioactive mutant. Furthermore, MS and MS2 experiments confirmed that X. cabanillasii produces the same derivatives as X. budapestensis. The medicament potential of 10-fold concentrated supernatant of induced fcl promoter exchanged X. cabanillasii was also assessed in dental root canals. Calcium hydroxide paste, or chlorhexidine gel, or fabclavine-rich supernatant was applied to root canals. Fabclavine-rich supernatant exhibited the highest inactivation efficacy of ≥3 log10 steps CFU reduction, followed by calcium hydroxide paste (≤2 log10 step). The mean percentage of E. faecalis-free dental root canals after treatment was 63.6, 45.5, and 18.2% for fabclavine, calcium hydroxide, and chlorhexidine, respectively. Fabclavine in liquid form or preferably as a paste or gel formulation is a promising alternative intracanal medicament.