Investigation of whole body extract metabolites of Lucilia sericata larvae and potential antibacterial effects

Abstract

Aim: Complementary medicinal techniques have gainedfocus by modern medicine, recently. Maggot Debridement Therapy is a widely-used method worldwide. It is especially recommended for chronic wounds, and has serious advantages such as low cost, easily-applicability and rare adverse effects, but its effect mechanisms remains unclear. The aim of this study is to detect components and to investigate potential antibacterial effects of whole body extract metabolites of Lucilia sericata larvae.

Material anf Methods: Due to potential antibacterial effects, agar well diffusion and flowcytometry methods were used against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Enterococcus faecalis to do evaluation on whole body extracts of previously-cloned maggots in specialized climate room. After this antibacterial effect evaluation, 2-D PAGE analysis was done for protein investigation.

Results: Inhibition zones were observed for S.aureus (16mm), E.coli (22mm) and E.faecalis (14mm), but for P.aeruginosa, the extract could not provide any inhibiton zone. In flow cytometry, different killing rates were detected in different extract dilutions, and for the lowest (1/64) dilution, killing rates were 51.9%, 75%, 80% and 98.7% for P.aeruginosa, E.faecalis, E.coli and S.aureus, respectively. 2-D PAGE showed various proteins with different molercular mass (<10-260kDa) and pI (3-9).

Conclusion: Antibacterial effects of maggot whole body extracts on tested strains are obviously detected. Many protein spots with widely variable molecular mass and isoelectric points were observed. As a result, this antibacterial effects may be caused by these proteins, but it is necessary that these proteins must be further evaluated via mass spectrometry and protein databases.

Keywords

• Lucilia sericata; Chronic Wound Care; Larval Debridement; Biosurgery; Maggot Debridement Therapy

Lucilia sericata larvalarının tüm vücut ekstrakt metabolitlerinin araştırılması ve potensiyel antibakteriyel etkileri

Abstract

Amaç: Yakın dönemde, tamamlayıcı tıp uygulamaları modern tıbbın ilgi alanına girmiştir. Maggot Debritman Tedavisi dünya çapında yaygın olarak kullanılan bir yöntemdir. Bu yöntem, özellikle kronik yaraların tedavisinde tavsiye edilmektedir ve düşük maliyet, kolay uygulanabilirlik ve nadir yan etkiler gibi avantajları bulunmaktadır, ancak yöntemin etki mekanizması henüz tam olarak ortaya konulamamıştır. Bu çalışmanın amacı, Lucilia sericatalarvalarının tüm vücut ekstraktının metabolitlerini ortaya koymak ve bunların potensiyel antibakteriyel niteliğini araştırmaktır.

Gereç ve Yöntemler: Antibakteriyel etkinliği araştırmak için, önceden özel iklim odalarında üretilmiş larvalarının tüm vücut ekstraktları, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coliveEnterococcus faecalisbakterileri için agar difüzyon ve akan hücreölçer ile test edilmiştir. Antibakteriyel incelemeyi takiben, iki boyutlu elektroforez ile protein araştırılması yapılmıştır.

Bulgular: S.aureus (16mm), E.coli (22mm) ve E.faecalis (14mm) için inhibisyon alanı gözlenmiş ancak P.aeruginosaiçinalan oluşmamıştır. Hücreölçer ile farklı dilüsyonlarda farklı öldürme oranları gözlenmiş ve en düşük dilüsyonda (1/64), P.aeruginosa, E.faecalis, E.coliveS.aureusiçin sırasıyla%51,9, %75, %80ve %98,7 oranları alınmıştır. İki boyutlu elektroforezde farklı moleküler ağırlık (<10-260kDa) ve izoelektrik noktada (3-9) proteinler tespit edilmiştir.

Sonuç: Maggot ekstraklarının test edilen suşlar üzerine antibakteriyel etkisi net olarak gözlenmiştir. Farklı moleküler ağırlık ve izoelektrik noktada proteinler tespit edilmiştir. Antibakteriyel etkinin bu proteinler tarafından sağlanması muhtemel olsa da, proteinlerin kütle spektrometrisi ve protein veri bankaları ile ayrıca araştırılması gerekir.

Keywords

• Lucilia sericata; Kronik Yara Bakımı; Larval Debritman; Biyocerrahi; Maggot Debritman Tedavisi

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