Desiccation and cold storage of Galleria mellonella cadavers and effects on in vivo production of Steinernema carpocapsae

2013 ◽  
Vol 70 (6) ◽  
pp. 895-904 ◽  
Author(s):  
Xin Wang ◽  
Huan Wang ◽  
Qing-zhou Feng ◽  
Xi-yang Cui ◽  
Ri-yue Liu ◽  
...  
Keyword(s):  
Cold Storage ◽  
Galleria Mellonella ◽  
Steinernema Carpocapsae

INFLUÊNCIA DO SUBSTRATO E DA LUMINOSIDADE NA INFECÇÃO DE NEMATOIDES ENTOMOPATOGÊNICOS EM Galleria mellonella (LEPIDOPTERA: PYRALIDAE)

2018 ◽  
Vol 20 (2) ◽  
pp. 91-101
Author(s):  
Andressa Lima de Brida ◽  
Silvia Renata Siciliano Wilcken ◽  
Luis Garrigós Leite
Keyword(s):  
Galleria Mellonella ◽  
Steinernema Carpocapsae ◽  
Steinernema Feltiae ◽  
Lepidoptera Pyralidae

Nematoides entomopatogênicos (NEPs) são alternativas eficientes para o controle de pragas. O emprego de novas técnicas da produção in vivo, permite o progresso da tecnologia de formulação de bioinseticidas. O objetivo do trabalho, foi avaliar a influência da luminosidade e do substrato na capacidade de infecção de juvenis infectantes (JIs) de Steinernema brazilense IBCBn 06, Steinernema carpocapsae IBCBn 02, Steinernema feltiae IBCBn 47 e Heterorhabditis amazonensis IBCBn 24 em lagartas de Galleria mellonella (Lepidoptera: Pyralidae). O delineamento experimental foi inteiramente casualizado com quatro tratamentos e oito repetições. As parcelas, constituídas por placa de Petri com, substrato-areia e substrato-papel filtro, com e sem luminosidade, inoculados com suspensão de 1,5 mL contendo 400JIs e quatro lagartas de G. mellonella. O número de JIs foi quantificado após a mortalidade das lagartas. A taxa de infecção de JIs de S. carpocapsae IBCBn 02 e S. feltiae IBCBn 47 variaram de 2,14 a 3,28 e de 11,04 a 13,09 JIs/lagarta. O substrato-areia com e sem luminosidade permitiu a maior taxa de infeção dos JIs de S. brazilense IBCBn 06 de 7,86 e 9,44 JIs/lagarta, e 13,49 JIs/lagarta com luminosidade para H. amazonensis IBCBn 24. O substrato-areia, permite a maior taxa de infecção por JIs de NEPs.

Effect of recycling temperature on the infectivity of entomopathogenic nematodes

1997 ◽  
Vol 75 (12) ◽  
pp. 2137-2141 ◽  
Author(s):  
Ganpat B. Jagdale ◽  
Roger Gordon
Keyword(s):  
Entomopathogenic Nematodes ◽  
Galleria Mellonella ◽  
Steinernema Carpocapsae ◽  
Steinernema Feltiae ◽  
Wax Moth

Four strains of entomopathogenic nematodes were recycled in vivo for 2 years at temperatures ranging from 10 to 25 °C, then the infectivity of their infective juveniles was compared. Infectivity was examined by measuring LC50 values for wax moth (Galleria mellonella) larvae at bioassay temperatures ranging from 5 to 25 °C. Of the four strains examined, only the Umeå and NF strains of Steinernema feltiae that had been recycled at 10 °C infected and killed the insects at a bioassay temperature of 5 °C. The Steinernema carpocapsae All and Steinernema riobravis TX strains were infective at 10 °C only when the recycling temperature was ≤ 20 °C. The infectivity of the two strains of S. feltiae at 10 or 15 °C was compromised by propagating them at higher temperatures (20–25 °C). The Umeå strain of S. feltiae displayed an impaired capacity to infect hosts at higher temperatures (20–25 °C) when recycled at lower (≤ 15 °C) temperatures. The capacity of these nematodes to adjust to different recycling temperatures is discussed in relation to their infectivity in different field situations.

Source of trait deterioration in entomopathogenic nematodes Heterorhabditis bacteriophora and Steinernema carpocapsae during in vivo culture

Nematology ◽  
2006 ◽  
Vol 8 (3) ◽  
pp. 397-409 ◽  
Author(s):  
Anwar L. Bilgrami ◽  
Randy Gaugler ◽  
David I. Shapiro-Ilan ◽  
Byron J. Adams
Keyword(s):  
Heat Tolerance ◽  
Galleria Mellonella ◽  
Entomopathogenic Nematode ◽  
Heterorhabditis Bacteriophora ◽  
Steinernema Carpocapsae ◽  
Reproductive Capacity ◽  
Impaired Performance ◽  
Trait Deterioration ◽  
The Stability

Abstract The stability of traits important for biological control was studied in the entomopathogenic nematode-bacteria complexes Heterorhabditis bacteriophora and Steinernema carpocapsae. Five experimental lines of each species were subcultured for 20 serial passages in Galleria mellonella larvae to assess trait stability. Subculturing impaired performance of both H. bacteriophora and S. carpocapsae. Virulence, heat tolerance and fecundity deteriorated in all H. bacteriophora experimental lines, and four out of five experimental lines deteriorated in host-finding ability. All S. carpocapsae experimental lines deteriorated in heat tolerance and nictation, and four out of five experimental lines declined for reproductive capacity, whereas virulence declined in two experimental lines. Determination of whether trait deterioration was due to changes in nematode, bacteria, or both symbiotic partners was tested by exchanging nematodes or bacteria from control populations with nematodes or bacteria from the most deteriorated experimental lines and assessing trait recovery. The source of deterioration varied according to trait, but only the bacterial partner played a role in trait reductions for every trait and species, whereas the nematode was the main source only for S. carpocapsae nictation. These results emphasise the important role each symbiotic partner plays in the stability and expression of beneficial traits.

scholarly journals A Peptide Found in Human Serum, Derived from the C-Terminus of Albumin, Shows Antifungal Activity In Vitro and In Vivo

2020 ◽  
Vol 8 (10) ◽  
pp. 1627
Author(s):  
Tecla Ciociola ◽  
Pier Paolo Zanello ◽  
Tiziana D’Adda ◽  
Serena Galati ◽  
Stefania Conti ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Human Serum ◽  
Fungicidal Activity ◽  
Galleria Mellonella ◽  
Serum Proteins ◽  
Morphological Alterations ◽  
C Terminus ◽  
Different Sources

The growing problem of antimicrobial resistance highlights the need for alternative strategies to combat infections. From this perspective, there is a considerable interest in natural molecules obtained from different sources, which are shown to be active against microorganisms, either alone or in association with conventional drugs. In this paper, peptides with the same sequence of fragments, found in human serum, derived from physiological proteins, were evaluated for their antifungal activity. A 13-residue peptide, representing the 597–609 fragment within the albumin C-terminus, was proved to exert a fungicidal activity in vitro against pathogenic yeasts and a therapeutic effect in vivo in the experimental model of candidal infection in Galleria mellonella. Studies by confocal microscopy and transmission and scanning electron microscopy demonstrated that the peptide penetrates and accumulates in Candida albicans cells, causing gross morphological alterations in cellular structure. These findings add albumin to the group of proteins, which already includes hemoglobin and antibodies, that could give rise to cryptic antimicrobial fragments, and could suggest their role in anti-infective homeostasis. The study of bioactive fragments from serum proteins could open interesting perspectives for the development of new antimicrobial molecules derived by natural sources.

scholarly journals Hexapod Assassins’ Potion: Venom Composition and Bioactivity from the Eurasian Assassin Bug Rhynocoris iracundus

Biomedicines ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 819
Author(s):  
Nicolai Rügen ◽  
Timothy P. Jenkins ◽  
Natalie Wielsch ◽  
Heiko Vogel ◽  
Benjamin-Florian Hempel ◽  
...  
Keyword(s):  
Biomedical Applications ◽  
Galleria Mellonella ◽  
Systemic Reactions ◽  
Venom Composition ◽  
Assassin Bug ◽  
Molecular Components ◽  
First Time ◽  
Tissue Digestion

Assassin bug venoms are potent and exert diverse biological functions, making them potential biomedical goldmines. Besides feeding functions on arthropods, assassin bugs also use their venom for defense purposes causing localized and systemic reactions in vertebrates. However, assassin bug venoms remain poorly characterized. We collected the venom from the assassin bug Rhynocoris iracundus and investigated its composition and bioactivity in vitro and in vivo. It caused lysis of murine neuroblastoma, hepatoma cells, and healthy murine myoblasts. We demonstrated, for the first time, that assassin bug venom induces neurolysis and suggest that it counteracts paralysis locally via the destruction of neural networks, contributing to tissue digestion. Furthermore, the venom caused paralysis and melanization of Galleria mellonella larvae and pupae, whilst also possessing specific antibacterial activity against Escherichia coli, but not Listeria grayi and Pseudomonas aeruginosa. A combinatorial proteo-transcriptomic approach was performed to identify potential toxins responsible for the observed effects. We identified neurotoxic Ptu1, an inhibitory cystin knot (ICK) toxin homologous to ω-conotoxins from cone snails, cytolytic redulysins homologous to trialysins from hematophagous kissing bugs, and pore-forming hemolysins. Additionally, chitinases and kininogens were found and may be responsible for insecticidal and cytolytic activities. We demonstrate the multifunctionality and complexity of assassin bug venom, which renders its molecular components interesting for potential biomedical applications.

scholarly journals In Silico Predicted Antifungal Peptides: In Vitro and In Vivo Anti-Candida Activity

2021 ◽  
Vol 7 (6) ◽  
pp. 439
Author(s):  
Tecla Ciociola ◽  
Walter Magliani ◽  
Tiziano De Simone ◽  
Thelma A. Pertinhez ◽  
Stefania Conti ◽  
...  
Keyword(s):  
In Silico ◽  
Mammalian Cells ◽  
Galleria Mellonella ◽  
Ex Vivo ◽  
Consensus Sequence ◽  
In Silico Analysis ◽  
Significant Activity ◽  
Synthetic Antibody

It has been previously demonstrated that synthetic antibody-derived peptides could exert a significant activity in vitro, ex vivo, and/or in vivo against microorganisms and viruses, as well as immunomodulatory effects through the activation of immune cells. Based on the sequence of previously described antibody-derived peptides with recognized antifungal activity, an in silico analysis was conducted to identify novel antifungal candidates. The present study analyzed the candidacidal and structural properties of in silico designed peptides (ISDPs) derived by amino acid substitutions of the parent peptide KKVTMTCSAS. ISDPs proved to be more active in vitro than the parent peptide and all proved to be therapeutic in Galleria mellonella candidal infection, without showing toxic effects on mammalian cells. ISDPs were studied by circular dichroism spectroscopy, demonstrating different structural organization. These results allowed to validate a consensus sequence for the parent peptide KKVTMTCSAS that may be useful in the development of novel antimicrobial molecules.

scholarly journals In vivo transcriptome analysis provides insights into host-dependent expression of virulence factors by Yersinia entomophaga MH96, during infection of Galleria mellonella

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Amber R Paulson ◽  
Maureen O’Callaghan ◽  
Xue-Xian Zhang ◽  
Paul B Rainey ◽  
Mark R H Hurst
Keyword(s):  
Galleria Mellonella ◽  
Functional Enrichment ◽  
Natural Environments ◽  
Insecticidal Toxin ◽  
Heat Stable ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Cell Densities

Abstract The function of microbes can be inferred from knowledge of genes specifically expressed in natural environments. Here, we report the in vivo transcriptome of the entomopathogenic bacterium Yersinia entomophaga MH96, captured during initial, septicemic, and pre-cadaveric stages of intrahemocoelic infection in Galleria mellonella. A total of 1285 genes were significantly upregulated by MH96 during infection; 829 genes responded to in vivo conditions during at least one stage of infection, 289 responded during two stages of infection, and 167 transcripts responded throughout all three stages of infection compared to in vitro conditions at equivalent cell densities. Genes upregulated during the earliest infection stage included components of the insecticidal toxin complex Yen-TC (chi1, chi2, and yenC1), genes for rearrangement hotspot element containing protein yenC3, cytolethal distending toxin cdtAB, and vegetative insecticidal toxin vip2. Genes more highly expressed throughout the infection cycle included the putative heat-stable enterotoxin yenT and three adhesins (usher-chaperone fimbria, filamentous hemagglutinin, and an AidA-like secreted adhesin). Clustering and functional enrichment of gene expression data also revealed expression of genes encoding type III and VI secretion system-associated effectors. Together these data provide insight into the pathobiology of MH96 and serve as an important resource supporting efforts to identify novel insecticidal agents.

Exploring Galleria mellonella larval model to evaluate antibacterial efficacy of Cecropin A (1-7)- Melittin against multi-drug resistant enteroaggregative Escherichia coli

2021 ◽  
Author(s):  
Jess Vergis ◽  
S V S Malik ◽  
Richa Pathak ◽  
Manesh Kumar ◽  
Nitin V Kurkure ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Galleria Mellonella ◽  
In Vivo Model ◽  
Drug Resistant ◽  
Physiological Concentration ◽  
Microbial Drug Resistance ◽  
Cecropin A ◽  
Screening And Identification

Abstract High throughput in vivo laboratory models is need for screening and identification of effective therapeutic agents to overcome microbial drug-resistance. This study was undertaken to evaluate in vivo antimicrobial efficacy of short-chain antimicrobial peptide- Cecropin A (1–7)-Melittin (CAMA) against three multi- drug resistant enteroaggregative Escherichia coli (MDR-EAEC) field isolates in a Galleria mellonella larval model. The minimum inhibitory concentration (MIC; 2.0 mg/L) and minimum bactericidal concentration (MBC; 4.0 mg/L) of CAMA were determined by microdilution assay. CAMA was found to be stable at high temperatures, physiological concentration of cationic salts and proteases; safe with sheep erythrocytes, secondary cell lines and commensal lactobacilli at lower MICs; and exhibited membrane permeabilisation. In vitro time-kill assay revealed concentration- and time-dependent clearance of MDR-EAEC in CAMA-treated groups at 30 min. CAMA- treated G. mellonella larvae exhibited an increased survival rate, reduced MDR-EAEC counts, immunomodulatory effect and proved non-toxic which concurred with histopathological findings. CAMA exhibited either an equal or better efficacy than the tested antibiotic control, meropenem. This study highlights the possibility of G. mellonella larvae as an excellent in vivo model for investigating the host-pathogen interaction, including the efficacy of antimicrobials against MDR-EAEC strains.

scholarly journals Designing P. aeruginosa synthetic phages with reduced genomes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Diana P. Pires ◽  
Rodrigo Monteiro ◽  
Dalila Mil-Homens ◽  
Arsénio Fialho ◽  
Timothy K. Lu ◽  
...  
Keyword(s):  
Galleria Mellonella ◽  
Antibacterial Properties ◽  
Genetically Engineered ◽  
In Vivo Studies ◽  
Infection Model ◽  
Promising Therapeutic Approach ◽  
Genes Encoding ◽  
First Time

AbstractIn the era where antibiotic resistance is considered one of the major worldwide concerns, bacteriophages have emerged as a promising therapeutic approach to deal with this problem. Genetically engineered bacteriophages can enable enhanced anti-bacterial functionalities, but require cloning additional genes into the phage genomes, which might be challenging due to the DNA encapsulation capacity of a phage. To tackle this issue, we designed and assembled for the first time synthetic phages with smaller genomes by knocking out up to 48% of the genes encoding hypothetical proteins from the genome of the newly isolated Pseudomonas aeruginosa phage vB_PaeP_PE3. The antibacterial efficacy of the wild-type and the synthetic phages was assessed in vitro as well as in vivo using a Galleria mellonella infection model. Overall, both in vitro and in vivo studies revealed that the knock-outs made in phage genome do not impair the antibacterial properties of the synthetic phages, indicating that this could be a good strategy to clear space from phage genomes in order to enable the introduction of other genes of interest that can potentiate the future treatment of P. aeruginosa infections.

Emergence of ceftazidime/avibactam resistance in KPC-3-producing Klebsiella pneumoniae in vivo

2019 ◽  
Vol 74 (11) ◽  
pp. 3211-3216 ◽  
Author(s):  
Stephan Göttig ◽  
Denia Frank ◽  
Eleonora Mungo ◽  
Anika Nolte ◽  
Michael Hogardt ◽  
...  
Keyword(s):  
Combination Therapy ◽  
Klebsiella Pneumoniae ◽  
Selection Pressure ◽  
Galleria Mellonella ◽  
Phenotypic Characterization ◽  
Infection Model ◽  
Development Of Resistance ◽  
Time Kill

Abstract Objectives The β-lactam/β-lactamase inhibitor combination ceftazidime/avibactam is active against KPC-producing Enterobacterales. Herein, we present molecular and phenotypic characterization of ceftazidime/avibactam resistance in KPC-3-producing Klebsiella pneumoniae that emerged in vivo and in vitro. Methods Sequence analysis of blaKPC-3 was performed from clinical and in vitro-generated ceftazidime/avibactam-resistant K. pneumoniae isolates. Time–kill kinetics and the Galleria mellonella infection model were applied to evaluate the activity of ceftazidime/avibactam and imipenem alone and in combination. Results The ceftazidime/avibactam-resistant clinical K. pneumoniae isolate revealed the amino acid change D179Y in KPC-3. Sixteen novel mutational changes in KPC-3 among in vitro-selected ceftazidime/avibactam-resistant isolates were described. Time–kill kinetics showed the emergence of a resistant subpopulation under selection pressure with either imipenem or ceftazidime/avibactam. However, combined selection pressure with imipenem plus ceftazidime/avibactam prevented the development of resistance and resulted in bactericidal activity. Concordantly, the G. mellonella infection model revealed that monotherapy with ceftazidime/avibactam is prone to select for resistance in vivo and that combination therapy with imipenem results in significantly better survival. Conclusions Ceftazidime/avibactam is a valuable antibiotic against MDR and carbapenem-resistant Enterobacterales. Based on time–kill kinetics as well as an in vivo infection model we postulate a combination therapy of ceftazidime/avibactam and imipenem as a strategy to prevent the development of ceftazidime/avibactam resistance in KPC-producing Enterobacterales in vivo.

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