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International Journal of Systematic and... Oct 2022One motile, Gram-negative, non-spore-forming and rod-shaped symbiotic bacterium, strain UCH-936, was isolated from nematodes. Results of biochemical, physiological,...
One motile, Gram-negative, non-spore-forming and rod-shaped symbiotic bacterium, strain UCH-936, was isolated from nematodes. Results of biochemical, physiological, molecular and genomic analyses suggest that it represents a new species, which we propose to name sp. nov. Digital DNA-DNA hybridization shows that strain UCH-936 is more closely related to DSM 23513, but shares solely 50.5 % similarity, which is below the 70% cut-off value that delimits species boundaries in bacteria. Phylogenetic reconstructions using whole-genome sequences show that strain UCH-936 forms a unique clade, suggesting its novel and distinct taxonomic status again. Similarly, comparative genomic analyses shows that the virulence factor flagella-related gene , the type IV pili-related gene and the vibriobactin-related gene are present in the genome of strain UCH-936 but absent in the genomes of its closest relatives. Biochemically and physiologically, UCH-936 differs also from all closely related species. Therefore, sp. nov. is proposed as a new species with the type strain UCH-936 (CCCT 21.06=CCM 9188=CCOS 1991).
Topics: Animals; Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Nematoda; Photorhabdus; Phylogeny; RNA, Ribosomal, 16S; Rhabditoidea; Sequence Analysis, DNA; Virulence Factors
PubMed: 36223180
DOI: 10.1099/ijsem.0.005525 -
Frontiers in Microbiology 2022The importance of and symbionts to their respective and nematode hosts is that they not only contribute to their entomopathogenicity but also to their fecundity... (Review)
Review
The importance of and symbionts to their respective and nematode hosts is that they not only contribute to their entomopathogenicity but also to their fecundity through the production of small molecules. Thus, this mini-review gives a brief introductory overview of these nematophilic bacteria. Specifically, their type species, nematode hosts, and geographic region of isolations are tabulated. The use of nucleotide sequence-based techniques for their species delineation and how pangenomes can improve this are highlighted. Using the - association as an example, the bacterium-nematode lifecycle is visualized with an emphasis on the role of bacterial biomolecules. Those currently in drug development are discussed, and two potential antimalarial lead compounds are highlighted. Thus, this mini-review tabulates forty-eight significant nematophilic bacteria and visualizes the ecological importance of their biomolecules. It further discusses three of these biomolecules that are currently in drug development. Through it, one is introduced to and bacteria, their natural production of biomolecules in the nematode-bacterium lifecycle, and how these molecules are useful in developing novel therapies.
PubMed: 36187939
DOI: 10.3389/fmicb.2022.993688 -
Experimental Parasitology Nov 2022The black cutworm (BCW), Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), is one of the destructive cutworm species. Black cutworm is a highly polyphagous pest that...
Biocontrol potential of cell suspensions and cell-free superntants of different Xenorhabdus and Photorhabdus bacteria against the different larval instars of Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae).
The black cutworm (BCW), Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), is one of the destructive cutworm species. Black cutworm is a highly polyphagous pest that feeds on more than 30 plants, many of which are of economic importance such as maize, sugar beet, and potato. The control of BCW larvae relies heavily on the application of synthetic insecticides which have a detrimental impact on human health and the natural environment. In addition, increasing insecticide resistance in many insect species requires a novel and sustainable approach to controlling insect pests. The endosymbionts of entomopathogenic nematodes (EPNs) (Xenorhabdus and Phorohabdus spp.) represent a newly emerging green approach to controlling a wide range of insect pests. In the current study, the oral and contact efficacy of cell suspension (4 × 10 cells ml) and cell-free supernatants of different symbiotic bacteria (X. nematophilai, X. bovienii, X. budapestensis, and P. luminescent subsp. kayaii) were evaluated against the mixed groups of 1st-2nd and 3rd-4th instars larvae of BCW under controlled conditions. The oral treatment of the cell suspension and cell-free supernatants resulted in higher mortality rates than contact treatments. In general, larval mortality was higher in the 1st-2nd instar larvae than in the 3rd-4th instar larvae. The highest (75%) mortality was obtained from the cell suspension of X. budapestensis. The results indicated that the oral formulations of the cell suspension and cell-free supernatants of bacterial strains may have a good control potential against the 1st-2nd larvae BCW. However, the efficacy of the cell suspension and cell-free supernatants of tested bacterial strains should be further evaluated under greenhouse and field conditions.
Topics: Animals; Humans; Larva; Xenorhabdus; Photorhabdus; Insecticides; Moths; Sugars; Pest Control, Biological
PubMed: 36179855
DOI: 10.1016/j.exppara.2022.108394 -
Nature Oct 2022Entomopathogenic nematodes are widely used as biopesticides. Their insecticidal activity depends on symbiotic bacteria such as Photorhabdus luminescens, which produces...
Entomopathogenic nematodes are widely used as biopesticides. Their insecticidal activity depends on symbiotic bacteria such as Photorhabdus luminescens, which produces toxin complex (Tc) toxins as major virulence factors. No protein receptors are known for any Tc toxins, which limits our understanding of their specificity and pathogenesis. Here we use genome-wide CRISPR-Cas9-mediated knockout screening in Drosophila melanogaster S2R+ cells and identify Visgun (Vsg) as a receptor for an archetypal P. luminescens Tc toxin (pTc). The toxin recognizes the extracellular O-glycosylated mucin-like domain of Vsg that contains high-density repeats of proline, threonine and serine (HD-PTS). Vsg orthologues in mosquitoes and beetles contain HD-PTS and can function as pTc receptors, whereas orthologues without HD-PTS, such as moth and human versions, are not pTc receptors. Vsg is expressed in immune cells, including haemocytes and fat body cells. Haemocytes from Vsg knockout Drosophila are resistant to pTc and maintain phagocytosis in the presence of pTc, and their sensitivity to pTc is restored through the transgenic expression of mosquito Vsg. Last, Vsg knockout Drosophila show reduced bacterial loads and lethality from P. luminescens infection. Our findings identify a proteinaceous Tc toxin receptor, reveal how Tc toxins contribute to P. luminescens pathogenesis, and establish a genome-wide CRISPR screening approach for investigating insecticidal toxins and pathogens.
Topics: Animals; Bacterial Toxins; Biological Control Agents; CRISPR-Cas Systems; Culicidae; Drosophila Proteins; Drosophila melanogaster; Fat Body; Gene Editing; Gene Knockdown Techniques; Hemocytes; Humans; Moths; Mucins; Pest Control, Biological; Phagocytosis; Photorhabdus; Repetitive Sequences, Amino Acid; Transgenes; Virulence Factors
PubMed: 36171290
DOI: 10.1038/s41586-022-05250-7 -
Journal of Invertebrate Pathology Oct 2022Photorhabdus insect related proteins A & B (PirA, PirB) from Photorhabdus and Xenorhabdus bacteria exhibit both oral and injectable toxicity against lepidopteran and...
Photorhabdus insect related proteins A & B (PirA, PirB) from Photorhabdus and Xenorhabdus bacteria exhibit both oral and injectable toxicity against lepidopteran and dipteran insect pest. The pirA, pirAt (encoding 6 N-terminal truncated PirA), pirB genes, pirA-pirB (with ERIC sequences), pirA-pirB-mERIC (modified pirA-pirB with mutated ERIC sequences) and polycistronic-pirAB were cloned and expressed in Escherichia coli. However, PirA protein was expressed in insoluble form and therefore the pirA gene was modified to produce PirAt. Moreover, pirA-pirB-mERIC, polycistronic-pirAB and co-transformed pirA/pirB genes were not expressed in the studied prokaryotic expression systems. None of the single purified proteins or mixtures of the individually expressed and purified proteins were toxic to mosquito larvae of Aedes aegypti and Culex quinquefasciatus. However, PirA-PirB protein mixtures purified from pirA-pirB operon plasmid were toxic to A. aegypti and C. quinquefasciatus larvae with LC values of 991 and 614 ng/ml, respectively. The presence of ERIC sequences between the two orfs of the pirA-pirB operon could help to obtain the proteins in biologically active form. Further, results confirm that PirA-PirB proteins of P. akhurstii subsp. akhurstii K-1 are binary insecticidal toxins and ERIC sequences could play an important role in expression of Pir proteins. Reports of biophysical characterization of individually purified PirAt, PirB and expressed PirA-PirB toxin mixture could provide the structural insight into these proteins.
Topics: Animals; Bacterial Proteins; Bacterial Toxins; Escherichia coli; Insect Proteins; Larva; Photorhabdus; Xenorhabdus
PubMed: 36167186
DOI: 10.1016/j.jip.2022.107829 -
Nature Microbiology Oct 2022Discovery of antibiotics acting against Gram-negative species is uniquely challenging due to their restrictive penetration barrier. BamA, which inserts proteins into the...
Discovery of antibiotics acting against Gram-negative species is uniquely challenging due to their restrictive penetration barrier. BamA, which inserts proteins into the outer membrane, is an attractive target due to its surface location. Darobactins produced by Photorhabdus, a nematode gut microbiome symbiont, target BamA. We reasoned that a computational search for genes only distantly related to the darobactin operon may lead to novel compounds. Following this clue, we identified dynobactin A, a novel peptide antibiotic from Photorhabdus australis containing two unlinked rings. Dynobactin is structurally unrelated to darobactins, but also targets BamA. Based on a BamA-dynobactin co-crystal structure and a BAM-complex-dynobactin cryo-EM structure, we show that dynobactin binds to the BamA lateral gate, uniquely protruding into its β-barrel lumen. Dynobactin showed efficacy in a mouse systemic Escherichia coli infection. This study demonstrates the utility of computational approaches to antibiotic discovery and suggests that dynobactin is a promising lead for drug development.
Topics: Animals; Anti-Bacterial Agents; Bacterial Outer Membrane Proteins; Escherichia coli; Escherichia coli Proteins; Gram-Negative Bacteria; Mice; Peptides; Phenylpropionates
PubMed: 36163500
DOI: 10.1038/s41564-022-01227-4 -
PloS One 2022Xenorhabdus and Photorhabdus can produce a variety of secondary metabolites with broad spectrum bioactivity against microorganisms. We investigated the antibacterial...
Genome analysis of secondary metabolite‑biosynthetic gene clusters of Photorhabdus akhurstii subsp. akhurstii and its antibacterial activity against antibiotic-resistant bacteria.
Xenorhabdus and Photorhabdus can produce a variety of secondary metabolites with broad spectrum bioactivity against microorganisms. We investigated the antibacterial activity of Xenorhabdus and Photorhabdus against 15 antibiotic-resistant bacteria strains. Photorhabdus extracts had strong inhibitory the growth of Methicillin-resistant Staphylococcus aureus (MRSA) by disk diffusion. The P. akhurstii s subsp. akhurstii (bNN168.5_TH) extract showed lower minimum inhibitory concentrations (MIC) and minimal bactericidal concentrations (MBC). The interaction between either P. akhurstii subsp. akhurstii (bNN141.3_TH) or P. akhurstii subsp. akhurstii (bNN168.5_TH) or P. hainanensis (bNN163.3_TH) extract in combination with oxacillin determined by checkerboard assay exhibited partially synergistic interaction with fractional inhibitory concentration index (FICI) of 0.53. Time-killing assay for P. akhurstii subsp. akhurstii (bNN168.5_TH) extract against S. aureus strain PB36 significantly decreased cell viability from 105 CFU/ml to 103 CFU/ml within 30 min (P < 0.001, t-test). Transmission electron microscopic investigation elucidated that the bNN168.5_TH extract caused treated S. aureus strain PB36 (MRSA) cell membrane damage. The biosynthetic gene clusters of the bNN168.5_TH contained non-ribosomal peptide synthetase cluster (NRPS), hybrid NRPS-type l polyketide synthase (PKS) and siderophore, which identified potentially interesting bioactive products: xenematide, luminmide, xenortide A-D, luminmycin A, putrebactin/avaroferrin and rhizomide A-C. This study demonstrates that bNN168.5_TH showed antibacterial activity by disrupting bacterial cytoplasmic membrane and the draft genome provided insights into the classes of bioactive products. This also provides a potential approach in developing a novel antibacterial agent.
Topics: Anti-Bacterial Agents; Methicillin-Resistant Staphylococcus aureus; Microbial Sensitivity Tests; Multigene Family; Oxacillin; Photorhabdus; Plant Extracts; Polyketide Synthases; Siderophores; Staphylococcus aureus; Xenorhabdus
PubMed: 36129957
DOI: 10.1371/journal.pone.0274956 -
Experimental Parasitology Nov 2022The fall webworm (FWW), Hyphantria cunea Drury (Lepidoptera: Erebidae), is an invasive and polyphagous insect pest of many economically important crops such as...
Insecticidal activities of the local entomopathogenic nematodes and cell-free supernatants from their symbiotic bacteria against the larvae of fall webworm, Hyphantriacunea.
The fall webworm (FWW), Hyphantria cunea Drury (Lepidoptera: Erebidae), is an invasive and polyphagous insect pest of many economically important crops such as hazelnuts, apple, and mulberry. Recently, there have been an increasing number of reports about the damaging activities of FWW from hazelnut growing areas of Turkey indicating that currently existing control methods fail to satisfy the expectations of growers. Entomopathogenic nematodes (EPNs) in the Steinernematidae and Heterorhabditidae (Nematoda: Rhabditida) families and the symbiotic bacteria they carry in their intestine have a great potential for the management of many agriculturally important pests. In this study, the symbiotic bacteria of local EPN species (Heterorhabditis bacteriophora AVB-15, Steinernema feltiae KCS-4S, and Steinernema bicornotum MGZ-4S) recovered from the central Anatolia region was characterized using recA gene region as Photorhabdus luminescens, Xenorhabdus bovienii and Xenorhabdus budapestensis. The contact (25, 50, 100, 200 IJs/Petri) and oral efficacies of the infective juveniles (IJs) (25, 50, 100, 200 IJs/leaf) of these EPN isolates determined on 3rd/4th instar larvae, and cell-free supernatants from the identified symbiotic bacteria were evaluated separately on the 3rd and 4th larval instars of FWW in Petri dish environment under laboratory conditions (25 ± 1 °C, 60% of RH). In the Petri dish bioassays of EPN species, the most pathogenic isolate at the 1st DAT and 4th DAT was S. feltiae which caused 50% mortality at the highest concentration (200 IJs/Petri) and the highest mortality rate (97.5%) were achieved at 4th DAT by H. bacteriophora AVB-15 isolate. Surprisingly, the mortality rates were generally higher at the lowest concentrations and 82.5% mortality were reached 4th DAT by S. bicornotum at the lowest concentration (25 IJs/leaf) in the leaf bioassays. Mortality rates were higher in both Petri dish and filter paper efficacies of cell-free supernatants at the 2nd DAT and the highest mortality (87.5%) was reached in the contact efficacy studies when applied X. bovienii KCS-4S strain. The results suggest that the tested EPN species and CFSs have good potential for biological control of the larvae of FWW and can contribute to the IPM programs of FWW. However, the efficacy of both IJs of EPNs and CFSs of their symbiotic bacteria on larvae of FWW requires further studies to verify their efficiency in the field.
Topics: Humans; Animals; Larva; Insecticides; Rhabditida; Photorhabdus; Moths; Pest Control, Biological
PubMed: 36116520
DOI: 10.1016/j.exppara.2022.108380 -
Pakistan Journal of Biological Sciences... Jun 2022<b>Background and Objective:</b> The housefly poses a threat to the public health of humans and domestic animals since it can carry and transmit pathogens....
<b>Background and Objective:</b> The housefly poses a threat to the public health of humans and domestic animals since it can carry and transmit pathogens. Despite there are many attempts to control this insect, most of them depend on conventional pesticides. Thus, the current study aimed to evaluate the efficacy of whole-cell suspension, cell-free supernatant and crude cells of the symbiotic bacteria <i>Photorhabdus</i> sp. and <i>Xenorhabdus</i> sp., as bio-control agents for housefly stages. <b>Materials and Methods:</b> The <i>Photorhabdus</i> sp. and <i>Xenorhabdus</i> sp., were isolated from the entomopathogenic nematodes, <i>Heterorhabditis indica</i> and <i>Steinernema feltiae</i>, respectively. The phenotypic, as well as the enzymatic characterizations of both bacteria, were determined. In addition, histopathological changes of the alimentary canal of <i>M. domestica</i> adults treated with whole-cell suspensions (at 3×10<sup>8 </sup>cells mL<sup></sup><sup>1</sup>) of both bacteria were carefully examined using transmission electron microscopy. <b>Results:</b> The results showed that both symbiotic bacteria significantly suppressed larvae, pupae and adults of <i>M. domestica</i>, particularly when they were applied as whole-cell suspensions. For example, the highest concentration of whole-cell suspension, cell-free supernatant and crude cells of <i>Photorhabdus</i> sp., induced larval mortalities by 94.7, 64.0 and 45.3%, while those of <i>Xenorhabdus</i> sp., induced larval mortalities by 58.7, 46.7 and 30.7% at 96 hrs, respectively. The results also showed that whole-cell suspensions of both symbiotic bacteria caused severe histopathological changes in the ultrastructure of the treated adults' alimentary canal. <b>Conclusion:</b> Both symbiotic bacteria can be effectively used, particularly the whole-cell suspension, as bio-control agents against the housefly either in the larval or adult stage.
Topics: Animals; Houseflies; Humans; Larva; Nematoda; Photorhabdus; Symbiosis; Xenorhabdus
PubMed: 36098165
DOI: 10.3923/pjbs.2022.586.601 -
Toxicon : Official Journal of the... Oct 2022Txp40 is a ubiquitous toxin from Xenorhabdus and Photorhabdus bacteria, exhibits insecticidal activity against a wide range of insect pests belonging to Lepidoptera and...
Txp40 is a ubiquitous toxin from Xenorhabdus and Photorhabdus bacteria, exhibits insecticidal activity against a wide range of insect pests belonging to Lepidoptera and Diptera orders. Initially, Txp40 affects midgut of the target insect and further damages some other tissues like fat bodies but the detailed mode of action is not known. Txp40 shares no significant sequence match to any proteins with known structure or function, suggesting that it is a novel type of insecticidal toxin. Here, we report purification, toxicity and biophysical characterization of the Txp40b toxin from X. nematophila (ATCC, 19061). The recombinant Txp40b was found toxic to Galleria mellonella larvae with LD of 30.42 ng larva. Circular dichroism spectroscopy revealed that purified Txp40b is an α-helix rich protein with a relatively lower melting temperature of 45 °C. In-silico model generated suggests two domain structure of Txp40b toxin. Detailed structural analysis of Txp40b will provide new insights about the mode of action and possibly it would illustrate a new domain and/or motif in the area of insecticidal proteins.
Topics: Animals; Bacterial Proteins; Insecta; Insecticides; Larva; Moths; Photorhabdus; Toxins, Biological; Xenorhabdus
PubMed: 36096207
DOI: 10.1016/j.toxicon.2022.09.003