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Pathogens (Basel, Switzerland) Mar 2023Symbiotic bacteria form a mutualistic relationship with nematodes and are pathogenic to many insect pests. They kill insects using various strategies to evade or...
Symbiotic bacteria form a mutualistic relationship with nematodes and are pathogenic to many insect pests. They kill insects using various strategies to evade or suppress their humoral and cellular immunity. Here we evaluate the toxic effects of these bacteria and their secondary metabolites on the survival and phenoloxidase (PO) activation of larvae using biochemical and molecular methods. The results show H06 and All treatments caused significant reductions in the number of larvae in a dose-dependent manner. Secondly, the immune system recognizes symbiotic bacteria at early and late stages of infection via the induction of C-type lectin. Live symbiotic bacteria significantly inhibit PO activity in whereas heat-treated bacteria strongly increase PO activity. Additionally, expression levels of four proPhenoloxidase genes following treatment with H06 and All were compared. We found that the expression levels of all proPhenoloxidase genes were significantly down-regulated at all-time points. Similarly, treatments of larvae with metabolites benzylideneacetone and oxindole significantly down-regulated the expression of the PPO gene and inhibited PO activity. However, the addition of arachidonic acid to metabolite-treated larvae restored the expression level of the PPO gene and increased PO activity. Our results provide new insight into the roles of symbiotic bacteria in countering the insect phenoloxidase activation system.
PubMed: 37111392
DOI: 10.3390/pathogens12040506 -
Microorganisms Mar 2023In bacteria, group-coordinated behavior such as biofilm formation or virulence are often mediated via cell-cell communication, a process referred to as quorum sensing...
In bacteria, group-coordinated behavior such as biofilm formation or virulence are often mediated via cell-cell communication, a process referred to as quorum sensing (QS). The canonical QS system of Gram-negative bacteria uses -acyl homoserine lactones (AHLs) as communication molecules, which are produced by LuxI-type synthases and sensed by cognate LuxR-type receptors. These receptors act as transcriptional regulators controlling the expression of specific genes. Some bacteria harbor LuxR-type receptors lacking a cognate LuxI-type synthases, designated as LuxR solos. Among many other LuxR solos, the entomopathogenic enteric bacterium harbors a SdiA-like LuxR solo containing an AHL signal-binding domain, for which a respective signal molecule and target genes have not been identified yet. Here we performed SPR analysis to demonstrate that SdiA acts as a bidirectional regulator of transcription, tightly controlling its own expression and the adjacent () gene in a gene supposed to be involved in the colonization of eukaryotes. Via qPCR we could further determine that in deletion mutant strains, is upregulated, indicating that SdiA negatively affects expression of . Furthermore, the Δ deletion mutant exhibited differences in biofilm formation and motility compared with the wild-type. Finally, using nanoDSF analysis we could identify putative binding ability of SdiA towards diverse AHLs, but also to plant-derived signals, modulating the DNA-binding capacity of SdiA, suggesting that this LuxR solo acts as an important player in interkingdom signaling between and plants.
PubMed: 37110313
DOI: 10.3390/microorganisms11040890 -
Journal of Invertebrate Pathology Jun 2023Locusts occasionally represent a danger in Africa despite intensive management measures, leading to severe yield loss and a commensurate loss of food and money. The...
Biochemical compositions and histopathology of the young and aged nymphs of the desert locust Schistocerca gregaria Forsskål (Orthoptera: Acrididae) affected by Photorhabdus luminescens (Enterobacterales: Morganellaceae).
Locusts occasionally represent a danger in Africa despite intensive management measures, leading to severe yield loss and a commensurate loss of food and money. The laboratory assessment of the toxicity of the Photorhabdus luminescens bacteria and its cell-free filtrate used Schistocerca gregaria nymphs in the second and fifth nymphs as test insects. Greater mortality was seen in locust nymphs of the second and fifth instars due to the high levels of toxicity produced by the bacterial suspension and its cell-free filtrate. The amounts of protein, fat, and carbohydrates in the treated locusts were drastically reduced. For the treated second and fifth instar nymphs of the desert locust, adverse effects on the muscular layers of the midgut and the muscles in the jumping legs were investigated.
Topics: Animals; Grasshoppers; Photorhabdus; Insecta
PubMed: 37068730
DOI: 10.1016/j.jip.2023.107922 -
Nature Apr 2023Endosymbiotic bacteria have evolved intricate delivery systems that enable these organisms to interface with host biology. One example, the extracellular contractile...
Endosymbiotic bacteria have evolved intricate delivery systems that enable these organisms to interface with host biology. One example, the extracellular contractile injection systems (eCISs), are syringe-like macromolecular complexes that inject protein payloads into eukaryotic cells by driving a spike through the cellular membrane. Recently, eCISs have been found to target mouse cells, raising the possibility that these systems could be harnessed for therapeutic protein delivery. However, whether eCISs can function in human cells remains unknown, and the mechanism by which these systems recognize target cells is poorly understood. Here we show that target selection by the Photorhabdus virulence cassette (PVC)-an eCIS from the entomopathogenic bacterium Photorhabdus asymbiotica-is mediated by specific recognition of a target receptor by a distal binding element of the PVC tail fibre. Furthermore, using in silico structure-guided engineering of the tail fibre, we show that PVCs can be reprogrammed to target organisms not natively targeted by these systems-including human cells and mice-with efficiencies approaching 100%. Finally, we show that PVCs can load diverse protein payloads, including Cas9, base editors and toxins, and can functionally deliver them into human cells. Our results demonstrate that PVCs are programmable protein delivery devices with possible applications in gene therapy, cancer therapy and biocontrol.
Topics: Animals; Humans; Mice; Cell Membrane; Eukaryotic Cells; Photorhabdus; CRISPR-Associated Protein 9; Toxins, Biological; Proteins; Drug Delivery Systems; Protein Transport
PubMed: 36991127
DOI: 10.1038/s41586-023-05870-7 -
Pathogens (Basel, Switzerland) Mar 2023Entomopathogenic bacteria are obligate symbionts of entomopathogenic nematode (EPN) species. These bacteria biosynthesize and release non-ribosomal-templated hybrid...
XENOFOOD-An Autoclaved Feed Supplement Containing Autoclavable Antimicrobial Peptides-Exerts Anticoccidial GI Activity, and Causes Bursa Enlargement, but Has No Detectable Harmful Effects in Broiler Cockerels despite In Vitro Detectable Cytotoxicity on LHM Cells.
Entomopathogenic bacteria are obligate symbionts of entomopathogenic nematode (EPN) species. These bacteria biosynthesize and release non-ribosomal-templated hybrid peptides (NR-AMPs), with strong, and large-spectral antimicrobial potential, capable of inactivating pathogens belonging to different prokaryote, and eukaryote taxa. The cell-free conditioned culture media (CFCM) of and efficiently inactivate poultry pathogens like , , and . To learn whether a bio-preparation containing antimicrobial peptides of origin with accompanying (in vitro detectable) cytotoxic effects could be considered a safely applicable preventive feed supplement, we conducted a 42-day feeding experiment on freshly hatched broiler cockerels. XENOFOOD (containing autoclaved , and cultures developed on chicken food) were consumed by the birds. The XENOFOOD exerted detectable gastrointestinal (GI) activity (reducing the numbers of the colony-forming units in the lower jejunum. No animal was lost in the experiment. Neither the body weight, growth rate, feed-conversion ratio, nor organ-weight data differed between the control (C) and treated (T) groups, indicating that the XENOFOOD diet did not result in any detectable adverse effects. We suppose that the parameters indicating a moderate enlargement of bursas of Fabricius (average weight, size, and individual bursa/spleen weight-ratios) in the XENOFOOD-fed group must be an indirect indication that the bursa-controlled humoral immune system neutralized the cytotoxic ingredients of the XENOFOOD in the blood, not allowing to reach their critical cytotoxic concentration in the sensitive tissues.
PubMed: 36986380
DOI: 10.3390/pathogens12030458 -
Biology Mar 2023Acute hepatopancreas necrosis disease (AHPND) has caused massive deaths of shrimp and has led to huge economic losses in aquaculture. (VP) carrying a plasmid encoding...
Acute hepatopancreas necrosis disease (AHPND) has caused massive deaths of shrimp and has led to huge economic losses in aquaculture. (VP) carrying a plasmid encoding binary toxins homologous to the insect-related (Pir) toxins is one of the main pathogens causing this disease. Previous studies have reported many immune-related genes of shrimp in response to this pathogenic bacteria. However, few studies have so far focused on the metabolic changes in upon VP infection. In the present study, comparative transcriptomic analysis was performed on the hepatopancreas of shrimp at different times during VP infection. Functional analyses on the differentially expressed genes (DEGs) during infection showed that pathways related to glucose, energy and amino acid metabolism, as well as nucleic acid synthesis, were obviously changed in the hepatopancreas after VP infection. Additionally, three signaling pathways, which could regulate metabolic processes, including HIF-1 signaling pathway, PI3K-Akt signaling pathway and NF-KappaB signaling pathway, also changed significantly. Collectively, these data reveal a close relationship between host metabolism processes and infection. The information will enrich our understanding of the interaction mechanism between the shrimp and .
PubMed: 36979109
DOI: 10.3390/biology12030417 -
Structure (London, England : 1993) May 2023Modification of the polyketide anthraquinone AQ-256 in the entomopathogenic Photorhabdus luminescens involves several O-methylations, but the biosynthetic gene cluster...
Modification of the polyketide anthraquinone AQ-256 in the entomopathogenic Photorhabdus luminescens involves several O-methylations, but the biosynthetic gene cluster antA-I lacks corresponding tailoring enzymes. We here describe the identification of five putative, highly homologous O-methyltransferases encoded in the genome of P. luminescens. Activity assays in vitro and deletion experiments in vivo revealed that three of them account for anthraquinone tailoring by producing three monomethylated and two dimethylated species of AQ-256. X-ray structures of all five enzymes indicate high structural and mechanistic similarity. As confirmed by structure-based mutagenesis, a conserved histidine at the active site likely functions as a general base for substrate deprotonation and subsequent methyl transfer in all enzymes. Eight complex structures with AQ-256 as well as mono- and dimethylated derivatives confirm the substrate specificity patterns found in vitro and visualize how single amino acid differences in the active-site pockets impact substrate orientation and govern site-specific methylation.
Topics: Methyltransferases; Methylation; Photorhabdus; Catalytic Domain; Anthraquinones
PubMed: 36963398
DOI: 10.1016/j.str.2023.03.001 -
Heliyon Mar 2023For decades, transcription of -operon was considered being constitutive. Therefore, this -operon has been used for measurements in non-specific bacterial luminescent...
For decades, transcription of -operon was considered being constitutive. Therefore, this -operon has been used for measurements in non-specific bacterial luminescent biosensors. Here, the expression of -operon under high temperature was studied. The expression was researched in the natural strain and in the heterologous system of . FV2201 bacterium was isolated from soil in the Moscow region (growth optimum 28 °C). We showed that its luminescence significantly increases when the temperature rises to 34 °C. The increase in luminescence is associated with an increase in the transcription of genes, which was confirmed by RT-PCR. The promoter of the -operon of the related bacterium ZM1 from the forests of Moldova, being cloned in the heterologous system of , is activated when the temperature rises from room temperature to 42 °C. When heat shock is caused by ethanol addition, transcription of -operon increases only in the natural strain of , but not in the heterologous system of cells. In addition, the activation of the -operon of persists in strains deficient in both the and genes. These results indicate the presence of sigma 32 and sigma 24 independent heat-shock-like mechanism of regulation of the -operon of in the heterologous system.
PubMed: 36950606
DOI: 10.1016/j.heliyon.2023.e14527 -
Journal of Invertebrate Pathology Jun 2023The grapevine moth, Lobesia botrana (Lepidoptera: Tortricidae), is a critical pest for vineyards and causes significant economic losses in wine-growing areas worldwide....
The grapevine moth, Lobesia botrana (Lepidoptera: Tortricidae), is a critical pest for vineyards and causes significant economic losses in wine-growing areas worldwide. Identifying and developing novel semiochemical cues (e.g. volatile bacterial compounds) which modify the ovipositional and trophic behaviour of L. botrana in vineyard fields could be a novel control alternative in viticulture. Xenorhabdus spp. and Photorhabdus spp. are becoming one of the best-studied bacterial species due to their potential interest in producing toxins and deterrent factors. In this study, we investigated the effect of the deterrent compounds produced by Xenorhabdus nematophila and Photorhabdus laumondii on the ovipositional moth behaviour and the larval feeding preference of L. botrana. Along with the in-vitro bioassays performed, we screened the potential use of 3 d cell-free bacterial supernatants and 3 and 5 d unfiltered bacterial ferments. In addition, we tested two application systems: (i) contact application of the bacterial compounds and (ii) volatile bacterial compounds application. Our findings indicate that the deterrent effectiveness varied with bacterial species, the use of bacterial cell-free supernatants or unfiltered fermentation product, and the culture times. Grapes soaked in the 3 d X. nematophila and P. laumondii ferments had ∼ 55% and ∼ 95% fewer eggs laid than the control, respectively. Likewise, the volatile compounds emitted by the 5 d P. laumondii fermentations resulted in ∼ 100% avoidance of L. botrana ovipositional activity for three days. Furthermore, both bacterial fermentation products have larval feeding deterrent effects (∼65% of the larva chose the control grapes), and they significantly reduced the severity of damage caused by third instar larva in treated grapes. This study provides insightful information about a novel bacteria-based tool which can be used as an eco-friendly and economical alternative in both organic and integrated control of L. botrana in vineyard.
Topics: Animals; Xenorhabdus; Photorhabdus; Moths; Larva; Vitis
PubMed: 36921888
DOI: 10.1016/j.jip.2023.107911 -
The Science of the Total Environment Jun 2023Larvae of the greater wax moth Galleria mellonella are common pests of beehives and commercial apiaries, and in more applied settings, these insects act as alternative...
Larvae of the greater wax moth Galleria mellonella are common pests of beehives and commercial apiaries, and in more applied settings, these insects act as alternative in vivo bioassays to rodents for studying microbial virulence, antibiotic development, and toxicology. In the current study, our aim was to assess the putative adverse effects of background gamma radiation levels on G. mellonella. To achieve this, we exposed larvae to low (0.014 mGy/h), medium (0.056 mGy/h), and high (1.33 mGy/h) doses of caesium-137 and measured larval pupation events, weight, faecal discharge, susceptibility to bacterial and fungal challenges, immune cell counts, activity, and viability (i.e., haemocyte encapsulation) and melanisation levels. The effects of low and medium levels of radiation were distinguishable from the highest dose rates used - the latter insects weighed the least and pupated earlier. In general, radiation exposure modulated cellular and humoral immunity over time, with larvae showing heightened encapsulation/melanisation levels at the higher dose rates but were more susceptible to bacterial (Photorhabdus luminescens) infection. There were few signs of radiation impacts after 7 days exposure, whereas marked changes were recorded between 14 and 28 days. Our data suggest that G. mellonella demonstrates plasticity at the whole organism and cellular levels when irradiated and offers insight into how such animals may cope in radiologically contaminated environments (e.g. Chornobyl Exclusion Zone).
Topics: Animals; Moths; Larva; Gamma Rays; Anti-Bacterial Agents; Virulence
PubMed: 36906041
DOI: 10.1016/j.scitotenv.2023.162742