Using the wax moth larva<i>Galleria mellonella</i>infection model to detect emerging bacterial pathogens

Hernández, Rafael; Hesse, Elze; Dowling, Andrea J.; Coyle, Nicola M.; Feil, Edward J.; Gaze, William H.; Vos, Michiel

doi:10.7717/peerj.6150

Cited by 33 publications

(45 citation statements)

References 40 publications

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“…Sampling types included suburban soils under grass or tree cover, as well as seven samples from an area contaminated with mine waste. A Galleria mellonella bioassay was used to screen soil microbial communities for pathogenic bacteria (Hernandez et al, 2019). Concentrated soil washes were directly injected into 10 Galleria each to assess percent mortality, after which they were incubated for 24 h at 37 C. A total of 29 (73%) out of 40 samples resulted in >5/10 Galleria mortality and were classified as potentially harbouring pathogenic bacteria ( Fig.…”

Section: Prevalence and Diversity Of Pathogenic Bacteria In Soils Sammentioning

confidence: 99%

Determining the prevalence, identity and possible origin of bacterial pathogens in soil

Ferraresso

Lawton

Bayliss

et al. 2020

Environmental Microbiology

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Soil biomes are vast, exceptionally diverse and crucial to the health of ecosystems and societies. Soils also contain an appreciable, but understudied, diversity of opportunistic human pathogens. With climate change and other forms of environmental degradation potentially increasing exposure risks to soilborne pathogens, it is necessary to gain a better understanding of their ecological drivers. Here we use the Galleria mellonella insect virulence model to selectively isolate pathogenic bacteria from soils in Cornwall (UK). We find a high prevalence of pathogenic soil bacteria with two genera, Providencia and Serratia, being especially common. Providencia alcalifaciens, P. rustigianii, Serratia liquefaciens and S. plymuthica strains were studied in more detail using phenotypic virulence and antibiotic resistance assays and whole-genome sequencing. Both genera displayed low levels of antibiotic resistance and antibiotic resistance gene carriage. However, Serratia isolates were found to carry the recently characterized metallo-β-lactamase blaSPR-1 that, although not conferring high levels of resistance in these strains, poses a potential risk of horizontal transfer to other pathogens where it could be fully functional. The Galleria assay can be a useful approach to uncover the distribution and identity of pathogenic bacteria in the environment, as well as uncover resistance genes with an environmental origin.

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Section: Prevalence and Diversity Of Pathogenic Bacteria In Soils Sammentioning

confidence: 99%

Determining the prevalence, identity and possible origin of bacterial pathogens in soil

Ferraresso

Lawton

Bayliss

et al. 2020

Environmental Microbiology

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“…Based on our knowledge of their different regulons, we attempted to further investigate the XRE-cupin TFs roles through two global phenotypic assays. We first took advantage of the Galleria mellonella infection model frequently used to assess overall bacterial fitness and virulence (Tsai et al, 2016, Andrejko et al, 2014, Hernandez et al, 2019. No significant difference could be observed in survival curves between larvae infected with wild-type and mutant strains ( Figure S3).…”

Section: Xre-cupin Tfs Regulate Specific Metabolic Pathwaysmentioning

confidence: 99%

Transcription inhibitors with XRE DNA-binding and cupin signal-sensing domains drive metabolic diversification in Pseudomonas

Trouillon

Ragno

Simon

et al. 2020

Preprint

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Transcription factors (TFs) are instrumental in the bacterial response to new environmental conditions. They can act as direct signal sensors and subsequently induce changes in gene expression leading to physiological adaptation. Here, by combining RNA-seq and DAP-seq, we studied a family of eight TFs in Pseudomonas aeruginosa. This family, encompassing TFs with XRE-like DNA-binding and cupin signal-sensing domains, includes the metabolic regulators ErfA, PsdR and PauR and five so far unstudied TFs. The genome-wide delineation of their regulons identified 39 regulatory interactions with genes mostly involved in metabolism. We found that the XRE-cupin TFs are inhibitors of their neighboring genes, forming local, functional units encoding proteins with functions in condition-specific metabolic pathways. The phylogenetic analysis of this family of regulators across the Pseudomonas genus revealed a wide diversity of such metabolic regulatory modules and identified species with potentially higher metabolic versatility. Numerous uncharacterized XRE-cupin TFs were found near metabolism-related genes, illustrating the need of further systematic characterization of transcriptional regulatory networks in order to better understand the mechanisms of bacterial adaptation to new environments.IMPORTANCEBacteria of the Pseudomonas genus, including the major human pathogen P. aeruginosa, are known for their complex regulatory networks and high number of transcription factors, which contribute to their impressive adaptive ability. However, even in the most studied species, most of the regulators are still uncharacterized. With the recent advances in high-throughput sequencing methods, it is now possible to fill this knowledge gap and help understanding how bacteria adapt and thrive in new environments. By leveraging these methods, we provide an example of a comprehensive analysis of an entire family of transcription factors and bring new insights into metabolic and regulatory adaptation in the Pseudomonas genus.

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“…To demonstrate the virulence of V. alginolyticus in aquaculture, sea bream ( Lutianus sanguineus ) and zebra fish ( Brachydanio rerio ) have been selected for V. alginolyticus infection models (Cai, Lu, Wu, & Jian, ; Chang et al, ). Additionally, wax moth ( Galleria mellonella ) caterpillars have successfully been utilized to study the pathogenicity of various human and animal bacterial pathogens, including Staphylococcus aureus , Pseudomonas aeruginosa , Listeria monocytogenes , Vibrio anguillarum and Vibrio cholerae (Hernandez et al, ; Jander, Rahme, & Ausubel, ; McMillan, Verner‐Jeffreys, Weeks, Austin, & Desbois, ; Mukherjee et al, ; Nuidate et al, ; Olsen, Watkins, Cantu, Beres, & Musser, ). Therefore, in this study, G. mellonella caterpillars were used as infection model according to the principle of replacement, reduction and refinement (3Rs; Tannenbaum & Bennett, ) to determine the pathogenicity of V. alginolyticus isolated from marine environments and diseased fish from southern Thailand.…”

Section: Introductionmentioning

confidence: 99%

Outer membrane protein A (OmpA) is a potential virulence factor of Vibrio alginolyticus strains isolated from diseased fish

Bunpa

Chaichana

Teng

et al. 2019

Journal of Fish Diseases

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Vibrio alginolyticus is one of the most serious causative agents of diseases in cultured marine fish and shellfish. However, the characteristics of virulence factors in pathogenic V. alginolyticus are poorly known. To gain insight into fish diseases caused by V. alginolyticus, we carried out two-dimensional gel electrophoresis (2-DE) combined with MALDI-TOF mass spectrometry to identify uniquely expressed proteins in the disease-causing V. alginolyticus. V. alginolyticus strains were isolated from marine environments and diseased fish obtained from southern Thailand. We identified seven unique proteins in the disease-causing V. alginolyticus strain. Among those, the outer membrane protein A (OmpA) had the strongest expression. Therefore, the function of this protein was further analysed. To investigate the role of OmpA protein, an in-frame deletion mutant of ompA was constructed using the homologous recombination method. Although the ompA mutant V. alginolyticus strain (ΔompA) grew normally, the mutant exhibited a significant defect in the swarming ability and the biofilm formation. Furthermore, Galleria mellonella larvae injected with the mutant bacteria had a significantly greater survival percentage than those injected with the wild-type strain, demonstrating that OmpA protein is required for the pathogenicity of V. alginolyticus. Together, this study suggests a potential target for vaccine development against pathogenic V. alginolyticus strain. K E Y W O R D SGalleria mellonella, MALDI-TOF mass spectrometry, ompA, two-dimensional gel electrophoresis, Vibrio alginolyticus How to cite this article: Bunpa S, Chaichana N, Teng JLL, et al. Outer membrane protein A (OmpA) is a potential virulence factor of Vibrio alginolyticus strains isolated from diseased fish. J Fish Dis. 2020;43:275-284. https ://doi.

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Using the wax moth larvaGalleria mellonellainfection model to detect emerging bacterial pathogens

Cited by 33 publications

References 40 publications

Determining the prevalence, identity and possible origin of bacterial pathogens in soil

Determining the prevalence, identity and possible origin of bacterial pathogens in soil

Transcription inhibitors with XRE DNA-binding and cupin signal-sensing domains drive metabolic diversification in Pseudomonas

Outer membrane protein A (OmpA) is a potential virulence factor of Vibrio alginolyticus strains isolated from diseased fish

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