Name: the insect galleria mellonella as a powerful infection model to investigate bacterial pathogenesis
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Description: Watch this Scientific Journal Video about The Insect Galleria mellonella as a Powerful Infection Model to Investigate Bacterial Pathogenesis at JoVE.com

Vi vil gjerne takke Elisabeth Guillemet, Christophe Buisson og Ludovic Bridoux for utmerket teknisk assistanse. Vi er sterkt i gjeld til Sylvie Salamitou og Sinda Fedhila for den første systemoppsettet.

1. Insekt Rearing Hele syklusen fra egg til siste instar larver varer ca 5 uker ved 25 ° C. Ett eller to ekstra uker for å få voksne sommerfuglene. <ol><li> Plasser minst 100 puppe eller nyfusjonerte voksen G. mellonella sommerfugler i en 5-liters wire-mesh buret. Mann sommerfugler høyden på 10 til 15 mm. Den voksne mannlige møll er beige med svakt lys og mørke markeringer. Kvinne sommerfugler måler rundt 20 mm. Kvinner er mørkere enn menn med en brun / grå farge. </li><...

<ol>
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Bruken av insekter og spesielt larvestadiet, som infeksjon modeller for flere patogener, blir hyppig. En modell av valget for noen aspekter er Drosophila (fly-modellen) brukes som både voksne og larvestadiet 1,2. Den lepidopteran insekt G. mellonella har også vært i hovedsak brukt til å analysere bakteriell virulens ved injeksjon. Fordelen med å tolerere høyere temperaturer (over 37 ° C) enn Drosophila (maks. 25 ° C) er viktig når pattedyr patogener er å bli undersøkt. Og...

<table border="1"><tbody><tr><td> Navnet på reagensen </td><td> Selskapet </td><td> Katalognummer </td><td> Kommentarer (valgfritt) </td></tr><tr><td> Voks og pollen </td><td> La Ruche Roanaise </td><td> 303000 </td><td> Noen honning produsent </td></tr><tr><td> Automatisert sprøytepumpe </td><td> KD Scientific </td><td> KDS 100 </td><td></td></tr><tr><td> Sprøyte 1 ml </td><td> Terumo </td><td> BS 01t </td><td></td></tr><tr><td> Nål 0,45 x 12 mm </td><td> Terumo </td><td> NN 2613R </td><td></td></tr><tr><td> Petriskål 5 cm </td><td> VWR </td><td> 89000-300 </td><td></td></tr><tr><td> Needle 30G, 25 mm sprøyte </td><td> Burkard Mfg Co Ltd </td><td> PDE0005 </td><td></td></tr></tbody></table>

the insect galleria mellonella as a powerful infection model to investigate bacterial pathogenesis

Studiet av bakteriell virulens krever ofte en egnet dyremodell. Pattedyr modeller av infeksjon er kostbare og kan reise etiske problemstillinger. Bruken av insekter som infeksjon modeller gir et verdifullt alternativ. Sammenlignet med andre ikke-virveldyr modell verter som nematoder, insekter har et relativt avansert system av antimikrobielle forsvar og er dermed mer sannsynlig å produsere informasjon som er relevant for pattedyr infeksjonen prosessen. Som pattedyr, insekter har et komplekst medfødte immunforsvaret en. Celler i hemolymph er i stand phagocytosing eller innkapsle mikrobielle inntrengere, og humorale responser inkluderer induserbare produksjon av lysozym og små antibakterielle peptider 2,3. I tillegg er analogier funnet mellom epitelceller av insekt larve midguts og tarmcellene pattedyr fordøyelsessystem. Endelig flere grunnleggende komponenter som er essensielle for bakteriell infeksjon prosessen som celleadhesjon, motstand motantimikrobielle peptider, vevsnedbrytning og tilpasning til oksidativt stress er sannsynlig å være viktig i både insekter og pattedyr en. Således er insekter polyvalente verktøy for identifikasjon og karakterisering av mikrobielle virulensfaktorer involvert i mammalske infeksjoner. Larver av større voks møll Galleria mellonella har vist seg å gi et nyttig innsikt i patogenesen av et bredt spekter av mikrobielle infeksjoner inkludert pattedyr fungal (Fusarium oxysporum, Aspergillus fumigatus, Candida albicans) og bakterielle patogener, så som Staphylococcus aureus, Proteus vulgaris , Serratia marcescens Pseudomonas aeruginosa, Listeria monocytogenes eller Enterococcus faecalis 4-7. Uavhengig av bakteriearter, resultater oppnådd med Galleria larver smittet av direkte innsprøytning gjennom cuticle konsekvent korrelerer med de av simiLar pattedyr studier: bakteriestammer som er svekket i pattedyr modeller viser lavere virulens i Galleria, og stammer som forårsaker alvorlige infeksjoner hos mennesker er også svært virulente i Galleria-modellen 8-11. Oral infeksjon av Galleria er mye mindre brukt og ytterligere forbindelser som spesifikke giftstoffer, er nødvendig for å nå dødelighet. G. mellonella larver presentere flere tekniske fordeler: de er relativt store (siste instar larver før forpupping er ca 2 cm lang og vekt 250 mg), dermed gjør injeksjon av definerte doser av bakterier, de kan bli oppdratt ved ulike temperaturer (20 ° C til 30 ° C) og infeksjon studier kan gjennomføres mellom 15 ° C til over 37 ° C 12,13, slik eksperimenter som etterligner et pattedyr miljø. I tillegg er insekt oppdrett enkelt og relativt billig. Infeksjon av larvene tillater overvåking bakteriell virulens på flere måter, including beregning av LD 50 14, måling av bakteriell overlevelse 15,16 og undersøkelse av infeksjonsprosessen 17. Her beskriver vi oppdra insekter, som dekker alle livsstadier av G. mellonella. Vi tilbyr en detaljert protokoll for smitte av to ruter av inokulering: muntlig og intra haemocoelic. Den bakterielle modellen som brukes i denne protokollen er Bacillus cereus, en Gram positiv patogen innblandet i gastrointestinal så vel som i andre alvorlige lokale eller systemiske opportunistiske infeksjoner 18,19.

Intra haemocoelic injeksjon av bakterier i G. mellonella påvist meget nyttige for identifisering av mange virulensfaktorer håndtere vevsskade og motstand mot medfødte immun faktorer av flere humane patogener. Som et eksempel, representerer figur 2A insektvoks dødelighet etter injeksjon av forskjellige doser av B. cereus bakterier (villtype og mutante stammer) 22. Fig. 2B representerer bakteriell overlevelse etter infisering av G. mellonella av <...

Watch this Scientific Journal Video about The Insect Galleria mellonella as a Powerful Infection Model to Investigate Bacterial Pathogenesis at JoVE.com

The Insect Galleria mellonella as a Powerful Infection Model to Investigate Bacterial Pathogenesis

Muntlig og intra haemocolic infeksjon av larver av større voks møll<em&gt; Galleria mellonella</em&gt; Er beskrevet. Dette insektet kan brukes til å studere virulensfaktorer av entomopathogenic samt pattedyr opportunistiske bakterier. Oppdrett av insekter, metoder for infeksjon og eksempler på<em&gt; In vivo</em&gt; Analyse er beskrevet.

Insektet<em&gt; Galleria mellonella</em&gt; Som en kraftig infeksjon Model for å etterforske Bakteriell Patogenese

The study of bacterial virulence often requires a suitable animal model. Mammalian models of infection are costly and may raise ethical issues. The use of ...

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Immunology and Infection

The Insect Galleria mellonella as a Powerful Infection Model to Investigate Bacterial Pathogenesis

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