scholarly journals UtilisingGalleria mellonella larvae for studying in vivo activity of conventional and novel antimicrobial agents

2020 ◽  
Vol 78 (8) ◽  
Author(s):  
Magdalena Piatek ◽  
Gerard Sheehan ◽  
Kevin Kavanagh
Keyword(s):  
Immune Response ◽  
Antimicrobial Agents ◽  
Galleria Mellonella ◽  
Antimicrobial Compounds ◽  
Insect Larvae ◽  
In Vivo Toxicity ◽  
Wide Range ◽  
Accuracy Of Results ◽  
Antibacterial And Antifungal

ABSTRACT The immune response of insects displays many structural and functional similarities to the innate immune response of mammals. As a result of these conserved features, insects may be used for evaluating microbial virulence or for testing the in vivo efficacy and toxicity of antimicrobial compounds and results show strong similarities to those from mammals. Galleria mellonella larvae are widely used in this capacity and have the advantage of being easy to use, inexpensive to purchase and house, and being free from the ethical and legal restrictions that relate to the use of mammals in these tests. Galleria mellonella larvae may be used to assess the in vivo toxicity and efficacy of novel antimicrobial compounds. A wide range of antibacterial and antifungal therapies have been evaluated in G. mellonella larvae and results have informed subsequent experiments in mammals. While insect larvae are a convenient and reproducible model to use, care must be taken in their use to ensure accuracy of results. The objective of this review is to provide a comprehensive account of the use of G. mellonella larvae for assessing the in vivo toxicity and efficacy of a wide range of antibacterial and antifungal agents.

scholarly journals Galleria mellonella: The Versatile Host for Drug Discovery, In Vivo Toxicity Testing and Characterising Host-Pathogen Interactions

Antibiotics ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1545
Author(s):  
Magdalena Piatek ◽  
Gerard Sheehan ◽  
Kevin Kavanagh
Keyword(s):  
Antimicrobial Agents ◽  
Galleria Mellonella ◽  
Toxicity Testing ◽  
In Vivo Model ◽  
Ethical Considerations ◽  
Food Preservatives ◽  
In Vivo Toxicity ◽  
Greater Wax Moth ◽  
Development Processes

Larvae of the greater wax moth, Galleria mellonella, are a convenient in vivo model for assessing the activity and toxicity of antimicrobial agents and for studying the immune response to pathogens and provide results similar to those from mammals. G. mellonella larvae are now widely used in academia and industry and their use can assist in the identification and evaluation of novel antimicrobial agents. Galleria larvae are inexpensive to purchase and house, easy to inoculate, generate results within 24–48 h and their use is not restricted by legal or ethical considerations. This review will highlight how Galleria larvae can be used to assess the efficacy of novel antimicrobial therapies (photodynamic therapy, phage therapy, metal-based drugs, triazole-amino acid hybrids) and for determining the in vivo toxicity of compounds (e.g., food preservatives, ionic liquids) and/or solvents (polysorbate 80). In addition, the disease development processes are associated with a variety of pathogens (e.g., Staphylococcus aureus, Listeria monocytogenes, Aspergillus fumigatus, Madurella mycotomatis) in mammals are also present in Galleria larvae thus providing a simple in vivo model for characterising disease progression. The use of Galleria larvae offers many advantages and can lead to an acceleration in the development of novel antimicrobials and may be a prerequisite to mammalian testing.

scholarly journals The Aspergillus fumigatus toxin fumagillin suppresses the immune response of Galleria mellonella larvae by inhibiting the action of haemocytes

Microbiology ◽  
2011 ◽  
Vol 157 (5) ◽  
pp. 1481-1488 ◽  
Author(s):  
John P. Fallon ◽  
Emer P. Reeves ◽  
Kevin Kavanagh
Keyword(s):  
Immune Response ◽  
Candida Albicans ◽  
Aspergillus Fumigatus ◽  
Immune Cells ◽  
Antimicrobial Agents ◽  
Galleria Mellonella ◽  
Subsequent Infection ◽  
Nadph Oxidase Complex ◽  
Cellular Immune

Larvae of Galleria mellonella are widely used to evaluate microbial virulence and to assess the in vivo efficacy of antimicrobial agents. The aim of this work was to examine the ability of an Aspergillus fumigatus toxin, fumagillin, to suppress the immune response of larvae. Administration of fumagillin to larvae increased their susceptibility to subsequent infection with A. fumigatus conidia (P = 0.0052). It was demonstrated that a dose of 2 µg fumagillin ml−1 reduced the ability of insect immune cells (haemocytes) to kill opsonized cells of Candida albicans (P = 0.039) and to phagocytose A. fumigatus conidia (P = 0.016). Fumagillin reduced the oxygen uptake of haemocytes and decreased the translocation of a p47 protein which is homologous to p47phox, a protein essential for the formation of a functional NADPH oxidase complex required for superoxide production. In addition, toxin-treated haemocytes showed reduced levels of degranulation as measured by the release of a protein showing reactivity to an anti-myeloperoxidase antibody (P<0.049) that was subsequently identified by liquid chromatography-MS analysis as prophenoloxidase. This work demonstrates that fumagillin suppresses the immune response of G. mellonella larvae by inhibiting the action of haemocytes and thus renders the larvae susceptible to infection. During growth of the fungus in the larvae, this toxin, along with others, may facilitate growth by suppressing the cellular immune response.

scholarly journals In VivoEfficacy of Anuran Trypsin Inhibitory Peptides against Staphylococcal Skin Infection and the Impact of Peptide Cyclization

2015 ◽  
Vol 59 (4) ◽  
pp. 2113-2121 ◽  
Author(s):  
U. Malik ◽  
O. N. Silva ◽  
I. C. M. Fensterseifer ◽  
L. Y. Chan ◽  
R. J. Clark ◽  
...  
Keyword(s):  
Antimicrobial Agents ◽  
Cyclic Peptide ◽  
Sequence Similarity ◽  
Infection Model ◽  
Content Type ◽  
Wide Range ◽  
Inhibitory Activities ◽  
The Impact

ABSTRACTStaphylococcus aureusis a virulent pathogen that is responsible for a wide range of superficial and invasive infections. Its resistance to existing antimicrobial drugs is a global problem, and the development of novel antimicrobial agents is crucial. Antimicrobial peptides from natural resources offer potential as new treatments against staphylococcal infections. In the current study, we have examined the antimicrobial properties of peptides isolated from anuran skin secretions and cyclized synthetic analogues of these peptides. The structures of the peptides were elucidated by nuclear magnetic resonance (NMR) spectroscopy, revealing high structural and sequence similarity with each other and with sunflower trypsin inhibitor 1 (SFTI-1). SFTI-1 is an ultrastable cyclic peptide isolated from sunflower seeds that has subnanomolar trypsin inhibitory activity, and this scaffold offers pharmaceutically relevant characteristics. The five anuran peptides were nonhemolytic and noncytotoxic and had trypsin inhibitory activities similar to that of SFTI-1. They demonstrated weakin vitroinhibitory activities againstS. aureus, but several had strong antibacterial activities againstS. aureusin anin vivomurine wound infection model. pYR, an immunomodulatory peptide fromRana sevosa, was the most potent, with complete bacterial clearance at 3 mg · kg−1. Cyclization of the peptides improved their stability but was associated with a concomitant decrease in antimicrobial activity. In summary, these anuran peptides are promising as novel therapeutic agents for treating infections from a clinically resistant pathogen.

scholarly journals Macrocolony of NDM-1 Producing Enterobacter hormaechei subsp. oharae Generates Subpopulations with Different Features Regarding the Response of Antimicrobial Agents and Biofilm Formation

Pathogens ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 49 ◽  
Author(s):  
Flávia Roberta Brust ◽  
Luana Boff ◽  
Danielle da Silva Trentin ◽  
Franciele Pedrotti Rozales ◽  
Afonso Luís Barth ◽  
...  
Keyword(s):  
Antimicrobial Agents ◽  
Galleria Mellonella ◽  
Polymyxin B ◽  
Multidrug Resistant ◽  
Biofilm Production ◽  
Depth Study ◽  
Enterobacter Hormaechei ◽  
Type 3

Enterobacter cloacae complex has been increasingly recognized as a nosocomial pathogen representing the third major Enterobacteriaceae species involved with infections. This study aims to evaluate virulence and antimicrobial susceptibility of subpopulations generated from macrocolonies of NDM-1 producing Enterobacter hormaechei clinical isolates. Biofilm was quantified using crystal violet method and fimbrial genes were investigated by PCR. Susceptibility of antimicrobials, alone and combined, was determined by minimum inhibitory concentration and checkerboard assays, respectively. Virulence and efficacy of antimicrobials were evaluated in Galleria mellonella larvae. Importantly, we verified that some subpopulations that originate from the same macrocolony present different biofilm production ability and distinct susceptibility to meropenem due to the loss of blaNDM-1 encoding plasmid. A more in-depth study was performed with the 798 macrocolony subpopulations. Type 3 fimbriae were straightly related with biofilm production; however, virulence in larvae was not statistically different among subpopulations. Triple combination with meropenem–rifampicin–polymyxin B showed in vitro synergistic effect against all subpopulations; while in vivo this treatment showed different efficacy rates for 798-1S and 798-4S subpopulations. The ability of multidrug resistant E. hormaechei isolates in generating bacterial subpopulations presenting different susceptible and virulence mechanisms are worrisome and may explain why these infections are hardly overcome.

scholarly journals Immune Response of Galleria mellonella against Human Fungal Pathogens

2018 ◽  
Vol 5 (1) ◽  
pp. 3 ◽  
Author(s):  
Nuria Trevijano-Contador ◽  
Oscar Zaragoza
Keyword(s):  
Immune Response ◽  
Fungal Pathogens ◽  
Histoplasma Capsulatum ◽  
Galleria Mellonella ◽  
Low Cost ◽  
Innate Immune ◽  
Human Pathogens ◽  
Lytic Enzymes ◽  
Wide Range ◽  
Great Specificity

In many aspects, the immune response against pathogens in insects is similar to the innate immunity in mammals. This has caused a strong interest in the scientific community for the use of this model in research of host–pathogen interactions. In recent years, the use of Galleria mellonella larvae, an insect belonging to the Lepidoptera order, has emerged as an excellent model to study the virulence of human pathogens. It is a model that offers many advantages; for example, it is easy to handle and establish in every laboratory, the larvae have a low cost, and they tolerate a wide range of temperatures, including human temperature 37 °C. The immune response of G. mellonella is innate and is divided into a cellular component (hemocytes) and humoral component (antimicrobial peptides, lytic enzymes, and peptides and melanin) that work together against different intruders. It has been shown that the immune response of this insect has a great specificity and has the ability to distinguish between different classes of microorganisms. In this review, we delve into the different components of the innate immune response of Galleria mellonella, and how these components manifest in the infection of fungal pathogens including Candida albicans, Aspergillus fumigatus, Cryptococcus neoformans, and Histoplasma capsulatum.

scholarly journals In Vitro and In Vivo Activity of Single and Dual Antimicrobial Agents Against KPC-producing Klebsiella pneumoniae

2017 ◽  
Vol 4 (suppl_1) ◽  
pp. S379-S379
Author(s):  
Farzad Moussavi ◽  
Sarath Nath ◽  
Daniel Abraham ◽  
David Landman ◽  
John Quale
Keyword(s):  
Antimicrobial Agents ◽  
Galleria Mellonella ◽  
Survival Rates ◽  
Polymyxin B ◽  
The Other ◽  
In Vivo Model ◽  
Serious Infections ◽  
Time Kill

Abstract Background Options for treatment of infections due to KPC-producing K. pneumoniae are limited, and combination therapy is often recommended. In this report, the in vitro and in vivo activity of potential therapeutic agents and combinations was assessed against four KPC-producing K. pneumoniae isolates. Methods Using clinically-relevant concentrations, time-kill experiments and the Galleria mellonella model of infection were used to examine the activity of polymyxin B, ceftazidime-avibactam, meropenem, rifampin, and amikacin alone and in combination. Four isolates of KPC-producing K. pneumoniae were studied, including two isolates that were resistant to polymyxin B and had ceftazidime-avibactam MICs of 8 µg/mL. The other two K. pneumoniae isolates were susceptible to polymyxin B and had lower MICs of ceftazidime-avibactam. Results Two isolates that were resistant to polymyxin B and with ceftazidime-avibactam MICs of 8 µg/mL were also resistant to amikacin and meropenem. When ceftazidime-avibactam was combined with either amikacin or meropenem, synergy was observed in vitro, and these combinations were associated with improved survival with the in vivo model. The other two K. pneumoniae isolates were susceptible to polymyxin B and had lower MICs of ceftazidime-avibactam. At concentrations four times the MIC, ceftazidime-avibactam had bactericidal activity in vitro; at one fourth the MIC, synergy was observed when combined with meropenem. Improved survival rates were observed with therapy with ceftazidime-avibactam, particularly when combined with a second agent for one isolate. In the in vivo model, polymyxin B with or without rifampin or meropenem, was ineffective against polymyxin B resistant strains. Conclusion Pending clinical studies, combining ceftazidime-avibactam with another agent (e.g., a carbapenem) should be encouraged when treating serious infections due to these pathogens, especially for isolates with ceftazidime-avibactam MICs near the susceptibility breakpoint. Disclosures All authors: No reported disclosures.

scholarly journals Recent Advances in the Use of Galleria mellonella Model to Study Immune Responses against Human Pathogens

2018 ◽  
Vol 4 (4) ◽  
pp. 128 ◽  
Author(s):  
Thais Pereira ◽  
Patrícia de Barros ◽  
Luciana Fugisaki ◽  
Rodnei Rossoni ◽  
Felipe Ribeiro ◽  
...  
Keyword(s):  
Immune Response ◽  
Immune Responses ◽  
Galleria Mellonella ◽  
Cellular Responses ◽  
Innate Immune ◽  
In Vivo Studies ◽  
Human Pathogens ◽  
Radical Production ◽  
Humoral Responses

The use of invertebrates for in vivo studies in microbiology is well established in the scientific community. Larvae of Galleria mellonella are a widely used model for studying pathogenesis, the efficacy of new antimicrobial compounds, and immune responses. The immune system of G. mellonella larvae is structurally and functionally similar to the innate immune response of mammals, which makes this model suitable for such studies. In this review, cellular responses (hemocytes activity: phagocytosis, nodulation, and encapsulation) and humoral responses (reactions or soluble molecules released in the hemolymph as antimicrobial peptides, melanization, clotting, free radical production, and primary immunization) are discussed, highlighting the use of G. mellonella as a model of immune response to different human pathogenic microorganisms.

scholarly journals UHPLC-ESI-QTOF-MS/MS-Based Molecular Networking Guided Isolation and Dereplication of Antibacterial and Antifungal Constituents of Ventilago denticulata

Antibiotics ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 606 ◽  
Author(s):  
Muhaiminatul Azizah ◽  
Patcharee Pripdeevech ◽  
Tawatchai Thongkongkaew ◽  
Chulabhorn Mahidol ◽  
Somsak Ruchirawat ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Herbal Medicine ◽  
Antibacterial Agents ◽  
Antimicrobial Agents ◽  
Antifungal Agents ◽  
Flavonoid Glycosides ◽  
Antimicrobial Compounds ◽  
Molecular Networking ◽  
Fragment Ions ◽  
Antibacterial And Antifungal

Ventilago denticulata is an herbal medicine for the treatment of wound infection; therefore this plant may rich in antibacterial agents. UHPLC-ESI-QTOF-MS/MS-Based molecular networking guided isolation and dereplication led to the identification of antibacterial and antifungal agents in V. denticulata. Nine antimicrobial agents in V. denticulata were isolated and characterized; they are divided into four groups including (I) flavonoid glycosides, rhamnazin 3-rhamninoside (7), catharticin or rhamnocitrin 3-rhamninoside (8), xanthorhamnin B or rhamnetin 3-rhamninoside (9), kaempferol 3-rhamninoside (10) and flavovilloside or quercetin 3-rhamninoside (11), (II) benzisochromanquinone, ventilatones B (12) and A (15), (III) a naphthopyrone ventilatone C (16) and (IV) a triterpene lupeol (13). Among the isolated compounds, ventilatone C (16) was a new compound. Moreover, kaempferol, chrysoeriol, isopimpinellin, rhamnetin, luteolin, emodin, rhamnocitrin, ventilagodenin A, rhamnazin and mukurozidiol, were tentatively identified as antimicrobial compounds in extracts of V. denticulata by a dereplication method. MS fragmentation of rhamnose-containing compounds gave an oxonium ion, C6H9O3+ at m/z 129, while that of galactose-containing glycosides provided the fragment ion at m/z 163 of C6H11O5+. These fragment ions may be used to confirm the presence of rhamnose or galactose in mass spectrometry-based analysis of natural glycosides or oligosaccharide attached to biomolecules, that is, glycoproteins.

Influence ofSteinernema feltiae(Filipjev) Wouts, Mracek, Gerdin and Bedding DD136 strain on the humoral and haemocytic responses ofGalleria mellonella(L.) larvae to selected bacteria

Parasitology ◽  
1985 ◽  
Vol 91 (2) ◽  
pp. 369-380 ◽  
Author(s):  
G. B. Dunphy ◽  
J. M. Webster
Keyword(s):  
Bacillus Subtilis ◽  
Galleria Mellonella ◽  
Steinernema Feltiae ◽  
Insect Larvae ◽  
Short Term ◽  
Xenorhabdus Nematophilus ◽  
Phosphate Buffered Saline

Examination of the short-term interaction of the haemocytes and lysozyme ofGalleria mellonellalarvae with the entomogenous nematodeSteinernema feltiaeDD136,in vitrorevealed that the nematodes did not reduce the adhesion ofBacillus subtilisorXenorhabdus nematophilussubsp.nematophilusto larval granulocytes or plasmatocytes. There was no evidence of humoral, sheath or cellular encapsulation ofS. feltiaein the haemolymphin vitroorin vivo. Compared with the phosphate-buffered saline-injected larvae the axenic nematodes did not alter the total or differential haemocyte counts during the initial 4 h of parasitism. The ability of the insect larvae to removeB. subtilisandX. nematophilusfrom the haemolymph was not influenced by axenicS. feltiae. The bacteria from the intestine of surface disinfected, monoxenically culturedS. feltiaeelevated the larval total haemocyte counts and damaged the haemocytes. The activity of larval lysozyme was not influenced by axenicS. feltiae.

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