Coronatin-1 isolated from entomopathogenic fungus Conidiobolus coronatus kills Galleria mellonella hemocytes in vitro and forms potassium channels in planar lipid membrane

Wieloch, Wioletta; Boguś, Mieczysława I.; Ligęza, Marta; Koszela-Piotrowska, Izabela; Szewczyk, Adam

doi:10.1016/j.toxicon.2011.07.007

Cited by 24 publications

(34 citation statements)

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“…The insecticidal activity of C. coronatus is attributed mainly to the presence of toxic metabolites, although it produces a broad range of enzymes, including proteinases, chitinases and lipases, which also play a crucial role in its pathogenicity (Prasertphon & Tanada, ; Boguś & Scheller, ; Freimoser et al ., ; Boguś et al ., , ; Wieloch et al ., ). Histopathological examination of G. mellonella larvae infected with C. coronatus does not reveal the presence of any nutrient depletion in dying insects (fat bodies were well preserved) or any damage to the vital internal organs of the host by fungal structures.…”

Section: Discussionmentioning

confidence: 97%

“…Previous investigations also report the isolation of a 30 kDa protein toxic for larvae of G. mellonella (Boguś & Scheller, ) and a serine protease of approximately 30–32 kDa with subtilisin properties (Bania et al ., ). The presence of proteo‐, chitino‐ and lipolytic enzymes is indicated in post‐incubation filtrates and in mycelia homogenates (Włóka & Boguś, ; Wieloch et al ., ; Boguś et al ., ). Our previous analyses (high‐perfomance liquid chomatography, liquid chomatography‐tandem mass spectrometry, electroelution, sodium dodecyl sulphate‐polyacrylamide gel electrophoresis) suggest that C. coronatus produces at least four insecticidal proteins: (i) coronatin‐1, a 36‐kDa protein forming potassium channels and showing both elastolytic and chitinolytic activities (Wieloch et al ., ); (ii) coronatin‐2, a 14.5‐kDa protein displaying no enzymatic activities (Boguś et al ., ); (iii) a 33–34‐kDa protein demonstrating elastolytic activity; and (iv) a 37‐kDa protein presenting no proteo‐, chitino‐ and lipolytic activities (Wieloch, ).…”

Section: Introductionmentioning

confidence: 99%

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Fungal infection causes changes in the number, morphology and spreading ability of Galleria mellonella haemocytes

et al. 2018

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The most effective and important strategy in the insect immune response is based on cellular reactions incorporating haemocytes. The present study uses Galleria mellonella (Lepidoptera: Pyralidae) as a host to study the pathogenesis caused by the entomopthoralean fungus Conidiobolus coronatus (Entomophthorales). Five types of haemocytes with different morphologies and behaviour are observed in the haemolymph of G. mellonella: granulocytes (GRs), plasmatocytes (PLs), spherulocytes (SPs), oenocytes (OEs) and prohaemocytes (PRs). During in vitro cultivation, three morphological subtypes of PLs are distinguished: flattened PLs, sun‐like PLs and oval PLs. In fresh smears of haemolymph observed under phase‐contrast microscopy, only flattened PLs are identified. No morphological changes are observed between fresh smears and in vitro cultures for GR, OE, SP and PR. Haemocytes cultured in vitro form a cellular network composed of PLs and GRs. Changes in the numbers, morphology and behaviour of haemocytes induced by fungal infection are compared with those observed in normally‐developing untreated larvae. Infection results in a significant drop in the number of haemocyte types. Fresh smears of haemocytes from mycosed larvae reveal malformed OEs, vacuolized PLs and GRs, as well as PLs with apoptotic blebs. Haemocytes from mycosed larvae incubated in vitro look similar, with degranulated GRs and vacuolized PLs forming microaggregations, as well as deformed OEs; only the SPs remain unharmed. Fungal infection impairs the ability of haemocytes to attach and spread on the culture dish. The actin cytoskeleton of haemocytes from mycosed larvae appear disorganized.

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Section: Discussionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

Fungal infection causes changes in the number, morphology and spreading ability of Galleria mellonella haemocytes

et al. 2018

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“…It seems that antimicrobial properties of alcohols and other cuticular compounds, together with the innate immunity mechanisms, efficiently protect M. domestica from the infection by common pathogens inhabiting similar habitats as those of the house fly. In contrast, the same compounds fail to protect adult flies against the entomopathogenic fungus C. coronatus 'equipped' with effective weaponry composed of proteolytic, chitinolytic and lipolytic enzymes that degrade the insect cuticle, as well as toxic metabolites that disable the victim's immune system (Wieloch et al, 2011). Similarly, Kontoyiannis and Lewis (Kontoyiannis and Lewis, 2010) reported high mortality of fruit flies infected with other Zygomycetes representatives -Rhizopus, Mucor and Cunninghamella species -contrasting with the resistance of Drosophila to Aspergillus and Candida species.…”

Section: Discussionmentioning

confidence: 99%

“…Assuming that adult flies are licking all substrates, it seems probable that during 18h of exposition to fungal colonies, insects ingest fungal spores, which quickly germinate inside the gut. Growing hyphae release toxic metabolites (Samborski, 2000;Boguś and Scheller, 2002;Wieloch et al, 2011), which may promptly kill the flies. Whether larvae burrowing in the fungal colony during the exposure ingest it as well remains unknown.…”

Section: Discussionmentioning

confidence: 99%

The antimicrobial activity of the alcohols fromMusca domestica

Gołębiowski

Dawgul

Kamysz

et al. 2012

Journal of Experimental Biology

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SUMMARYInformation on the stimulatory and inhibitory effects of cuticular alcohols on growth and virulence of insecticidal fungi is unavailable. Therefore, we set out to describe the content of cuticular and internal alcohols in the body of housefly larvae, pupae, males and females. The total cuticular alcohols in larvae, males and females of Musca domestica were detected in comparable amounts (4.59, 3.95 and 4.03gg -1 insect body, respectively), but occurred in smaller quantities in pupae (2.16gg -1). The major free alcohol in M. domestica larvae was C 12:0 (70.4%). Internal alcohols of M. domestica larvae were not found. Among cuticular pupae alcohols, C 12:0 (31.0%) was the most abundant. In the internal lipids of pupae, only five alcohols were identified in trace amounts. The most abundant alcohol in males was C 24:0 (57.5%). The percentage content of cuticular C 24:0 in males and females (57.5 and 36.5%, respectively) was significantly higher than that of cuticular lipids in larvae and pupae (0.9 and 5.6%, respectively). , respectively). For isolated alcohols, antimicrobial activity against 10 reference strains of bacteria and fungi was determined. Individual alcohols showed approximately equal activity against fungal strains. C 14:0 was effective against gram-positive bacteria, whereas gram-negative bacteria were resistant to all tested alcohols. Mixtures of alcohols found in cuticular lipids of larvae, pupae, males and females of M. domestica generally presented higher antimicrobial activity than individual alcohols. In contrast, crude extracts containing both cuticular and internal lipids showed no antifungal activity against the entomopathogenic fungus Conidiobolus coronatus, which efficiently kills adult house flies. Only two alcohols were present in the internal lipids of males in trace amounts

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“…These enzymes enable Conidiobolus to digest human tissue, growing by extension from nasal submucosa through the facial soft and bony tissue (Gugnani 1992). Metabolites toxic to insects have also been identified but because the fungus is highly lethal to arthropods and only of moderate virulence in mammals, it remains to be seen whether these metabolites play any role in human infection (Wieloch et al 2011).…”

Section: Entomophthoromycotamentioning

confidence: 99%