scholarly journals A novel restraint device for injection of Galleria mellonella larvae that minimises the risk of accidental operator needle stick injury

2016 ◽  
Author(s):  
James P Dalton ◽  
Benedict Uy ◽  
Simon Swift ◽  
Siouxsie Wiles
Keyword(s):  
Antimicrobial Agents ◽  
Galleria Mellonella ◽  
Cost Effective ◽  
Manual Dexterity ◽  
Needle Stick Injury ◽  
Drug Resistant ◽  
Highly Pathogenic ◽  
Needle Stick ◽  
Risk Of Injury ◽  
Pathogenic Microbes

Larvae of the insect Galleria mellonella are increasingly being used for studying pathogenic microbes and their virulence mechanisms, and as a rapid model for screening novel antimicrobial agents. The larvae (waxworms) are most frequently infected by injection of pathogenic organisms into the haemocoel through the insect's prolegs. The mostly widely used method for restraining the waxworms for injection is by grasping them between the operator's fingers, which puts the operator at risk of needle stick injury, an important consideration when working with highly pathogenic and/or drug-resistant microorganisms. While use of a stab proof glove can reduce this risk of injury, it does so at the loss of manual dexterity and speed, resulting in a more labour-intensive and cumbersome assay. We describe a simple cost effective device (the so-called 'Galleria Grabber') for restraining waxworms for injection that keeps the operator's fingers clear of the needle thus reducing the risk of injury.

scholarly journals Isolation and primary culture of Galleria mellonella hemocytes for infection studies

F1000Research ◽  
2021 ◽  
Vol 9 ◽  
pp. 1392
Author(s):  
Nicola J. Senior ◽  
Richard W. Titball
Keyword(s):  
Primary Culture ◽  
Alternative Model ◽  
Galleria Mellonella ◽  
Microbial Pathogens ◽  
Needle Stick Injury ◽  
Antimicrobial Compounds ◽  
Phagocytic Cells ◽  
Needle Stick ◽  
Research Institutes ◽  
Pre Treatment

Galleria mellonella larvae are increasingly used to study the mechanisms of virulence of microbial pathogens and to assess the efficacy of antimicrobials.  The G. mellonella model can faithfully reproduce many aspects of microbial disease which are seen in mammals, and therefore allows a reduction in the use of mammals. The model is now being widely used by researchers in universities, research institutes and industry. An attraction of the model is the interaction between pathogen and host. Hemocytes are specialised phagocytic cells which resemble neutrophils in mammals and play a major role in the response of the larvae to infection. However, the detailed interactions of hemocytes with pathogens is poorly understood, and is complicated by the presence of different sub-populations of cells. We report here a method for the isolation of hemocytes from Galleria mellonella.  A needle-stick injury of larvae, before harvesting, markedly increased the recovery of hemocytes in the hemolymph. The majority of the hemocytes recovered were granulocyte-like cells. The hemocytes survived for at least 7 days in culture at either 28°C or 37°C. Pre-treatment of larvae with antibiotics did not enhance the survival of the cultured hemocytes. Our studies highlight the importance of including sham injected, rather than un-injected, controls when the G. mellonella model is used to test antimicrobial compounds. Our method will now allow investigations of the interactions of microbial pathogens with insect hemocytes enhancing the value of G. mellonella as an alternative model to replace the use of mammals, and for studies on hemocyte biology.

scholarly journals Isolation and primary culture of Galleria mellonella hemocytes for infection studies

F1000Research ◽  
2020 ◽  
Vol 9 ◽  
pp. 1392
Author(s):  
Nicola J. Senior ◽  
Richard W. Titball
Keyword(s):  
Primary Culture ◽  
Alternative Model ◽  
Galleria Mellonella ◽  
Microbial Pathogens ◽  
Needle Stick Injury ◽  
Antimicrobial Compounds ◽  
Phagocytic Cells ◽  
Needle Stick ◽  
Research Institutes ◽  
Pre Treatment

Galleria mellonella larvae are increasingly used to study the mechanisms of virulence of microbial pathogens and to assess the efficacy of antimicrobials.  The G. mellonella model can faithfully reproduce many aspects of microbial disease which are seen in mammals, and therefore allows a reduction in the use of mammals. The model is now being widely used by researchers in universities, research institutes and industry. An attraction of the model is the interaction between pathogen and host. Hemocytes are specialised phagocytic cells which resemble neutrophils in mammals and play a major role in the response of the larvae to infection. However, the detailed interactions of hemocytes with pathogens is poorly understood, and is complicated by the presence of different sub-populations of cells. We report here a method for the isolation of hemocytes from Galleria mellonella.  A needle-stick injury of larvae, before harvesting, markedly increased the recovery of hemocytes in the hemolymph. The majority of the hemocytes recovered were granulocyte-like cells. The hemocytes survived for at least 7 days in culture at either 28°C or 37°C. Pre-treatment of larvae with antibiotics did not enhance the survival of the cultured hemocytes. Our studies highlight the importance of including sham injected, rather than un-injected, controls when the G. mellonella model is used to test antimicrobial compounds. Our method will now allow investigations of the interactions of microbial pathogens with insect hemocytes enhancing the value of G. mellonella as an alternative model to replace the use of mammals, and for studies on hemocyte biology.

scholarly journals Detection of Carbapenemase-Producing Klebsiella pneumoniae and Escherichia coli Recovered from Clinical Specimens in Erbil City Kurdistan Region of Iraq

2019 ◽  
Vol 30 (2) ◽  
pp. 10
Author(s):  
Heshu Jalal Ahmed ◽  
Aryan R. Ganjo
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Molecular Detection ◽  
Antimicrobial Agents ◽  
Cost Effective ◽  
Drug Resistant ◽  
Diffusion Test ◽  
Detection Techniques ◽  
Carbapenem Resistant ◽  
Disk Test

Background: Carbapenems are usually the choice of antimicrobial agents in infections produced by Enterobacteriaceae bacteria-producing ESBL (extended spectrum β-lactamases). Carbapenemase production among clinical isolates of Enterobacteriaceae has been widely reported and Resistance to carbapenems group is generally due to production of Carbapenemases. Phenotypic determination and distinction of Carbapenemases in drug-resistant gram-negative is crucial for appropriate infection control. Materials and Methods: Carbapenemase production among Enterobacteriaceae isolates was identified phenotypically using a commercially available EDTA-combined disc diffusion test containing inhibitors to the various carbapenemase classes and Modified Hodge test (MHT). Results: A total of 98 Enterobacteriaceae isolates were included, 42(42.8%) were Multi-drug resistant (MDR), 27(27.5%) were XDR while 8(8.2%) exhibited pan-drug resistance (PDR). Of the 74 isolates of Escherichia coli and 24 Klebsiella pneumoniae that were positive for carbapenemase production, 12 (16.2%) and 9 (37.5%) were Metallo beta-lactamase (MBL) producers respectively, Hence, the overall prevalence of carbapenemase-producing Escherichia coli and Klebsiella pneumoniae in this study were 47.3% and 87.5%. Conclusion: Carbapenemase-producing Enterobacteriaceae was indeed recognized in our hospitals. The EDTA-combination disk test was a rapid, cost-effective and suitable method which will be able to identify and distinguish the carbapenem-resistant bacterial isolates within the hospitals especially when molecular detection techniques are not available.

scholarly journals Design of a Broad-Range Bacteriophage Cocktail That ReducesPseudomonas aeruginosaBiofilms and Treats Acute Infections in Two Animal Models

2018 ◽  
Vol 62 (6) ◽  
Author(s):  
Francesca Forti ◽  
Dwayne R. Roach ◽  
Marco Cafora ◽  
Maria E. Pasini ◽  
David S. Horner ◽  
...  
Keyword(s):  
Antimicrobial Agents ◽  
Galleria Mellonella ◽  
Cost Effective ◽  
The United States ◽  
Multidrug Resistant ◽  
Bacterial Strains ◽  
Content Type ◽  
Clinical Strains ◽  
Phage Cocktail ◽  
The Individual

ABSTRACTThe alarming diffusion of multidrug-resistant (MDR) bacterial strains requires investigations on nonantibiotic therapies. Among such therapies, the use of bacteriophages (phages) as antimicrobial agents, namely, phage therapy, is a promising treatment strategy supported by the findings of recent successful compassionate treatments in Europe and the United States. In this work, we combined host range and genomic information to design a 6-phage cocktail killing several clinical strains ofPseudomonas aeruginosa, including those collected from Italian cystic fibrosis (CF) patients, and analyzed the cocktail performance. We demonstrated that the cocktail composed of four novel phages (PYO2, DEV, E215 and E217) and two previously characterized phages (PAK_P1 and PAK_P4) was able to lyseP. aeruginosaboth in planktonic liquid cultures and in biofilms. In addition, we showed that the phage cocktail could cure acute respiratory infection in mice and treat bacteremia in wax moth (Galleria mellonella) larvae. Furthermore, administration of the cocktail to larvae prior to bacterial infection provided prophylaxis. In this regard, the efficiency of the phage cocktail was found to be unaffected by the MDR or mucoid phenotype of the pseudomonal strain. The cocktail was found to be superior to the individual phages in destroying biofilms and providing a faster treatment in mice. We also found theGallerialarva model to be cost-effective for testing the susceptibility of clinical strains to phages, suggesting that it could be implemented in the frame of developing personalized phage therapies.

NMR Assisted Antimicrobial Peptide Designing: Structure Based Modifications and Functional Correlation of a Designed Peptide VG16KRKP

2020 ◽  
Vol 27 (9) ◽  
pp. 1387-1404 ◽  
Author(s):  
Karishma Biswas ◽  
Humaira Ilyas ◽  
Aritreyee Datta ◽  
Anirban Bhunia
Keyword(s):  
Antimicrobial Agents ◽  
Biological Properties ◽  
Structure Characterization ◽  
Drug Resistant ◽  
Functional Correlation ◽  
Naturally Occurring ◽  
High Resolution Nmr ◽  
Different Types ◽  
Reduced Activity ◽  
Poor Stability

Antimicrobial Peptides (AMPs), within their realm incorporate a diverse group of structurally and functionally varied peptides, playing crucial roles in innate immunity. Over the last few decades, the field of AMP has seen a huge upsurge, mainly owing to the generation of the so-called drug resistant ‘superbugs’ as well as limitations associated with the existing antimicrobial agents. Due to their resilient biological properties, AMPs can very well form the sustainable alternative for nextgeneration therapeutic agents. Certain drawbacks associated with existing AMPs are, however, issues of major concern, circumventing which are imperative. These limitations mainly include proteolytic cleavage and hence poor stability inside the biological systems, reduced activity due to inadequate interaction with the microbial membrane, and ineffectiveness because of inappropriate delivery among others. In this context, the application of naturally occurring AMPs as an efficient prototype for generating various synthetic and designed counterparts has evolved as a new avenue in peptide-based therapy. Such designing approaches help to overcome the drawbacks of the parent AMPs while retaining the inherent activity. In this review, we summarize some of the basic NMR structure based approaches and techniques which aid in improving the activity of AMPs, using the example of a 16-residue dengue virus fusion protein derived peptide, VG16KRKP. Using first principle based designing technique and high resolution NMR-based structure characterization we validate different types of modifications of VG16KRKP, highlighting key motifs, which optimize its activity. The approaches and designing techniques presented can support our peers in their drug development work.

Antimicrobial properties of actively purified secondary metabolites isolated from different marine organisms

2020 ◽  
Vol 21 ◽  
Author(s):  
Nilushi Indika Bamunu Arachchige ◽  
Fazlurrahman Khan ◽  
Young-Mog Kim
Keyword(s):  
Secondary Metabolites ◽  
Pathogenic Bacteria ◽  
Antimicrobial Agents ◽  
Bacterial Species ◽  
Antimicrobial Properties ◽  
Antimicrobial Effect ◽  
Cost Effective ◽  
Resistance Mechanisms ◽  
Marine Organisms ◽  
Antimicrobial Compounds

Background: The treatment of infection caused by pathogenic bacteria becomes one of the serious concerns globally. The failure in the treatment was found due to the exhibition of multiple resistance mechanisms against the antimicrobial agents. Emergence of resistant bacterial species has also been observed due to prolong treatment using conventional antibiotics. To combat these problems, several alternative strategies have been employed using biological and chemically synthesized compounds as antibacterial agents. Marine organisms considered as one of the potential sources for the isolation of bioactive compounds due to the easily available, cost-effective, and eco-friendly. Methods: The online search methodology was adapted for the collection of information related to the antimicrobial properties of marine-derived compounds. These compound has been isolated and purified by different purification techniques, and their structure also characterized. Furthermore, the antibacterial activities have been reported by using broth microdilution as well as disc diffusion assays. Results: The present review paper describes the antimicrobial effect of diverse secondary metabolites which are isolated and purified from the different marine organisms. The structural elucidation of each secondary metabolite has also been done in the present paper, which will help for the in silico designing of the novel and potent antimicrobial compounds. Conclusion: A thorough literature search has been made and summarizes the list of antimicrobial compounds that are isolated from both prokaryotic and eukaryotic marine organisms. The information obtained from the present paper will be helpful for the application of marine compounds as antimicrobial agents against different antibiotic-resistant human pathogenic bacteria.

Potential Antimicrobial Peptides Elucidation From The Marine Bacteria

2020 ◽  
Vol 18 ◽  
Author(s):  
Adyasa Barik ◽  
Pandiyan Rajesh ◽  
Manthiram Malathi ◽  
Vellaisamy Balasubramanian
Keyword(s):  
Antimicrobial Peptide ◽  
Marine Bacteria ◽  
Unnatural Amino Acids ◽  
Wide Spectrum ◽  
Drug Resistant ◽  
Medical Field ◽  
The World ◽  
Pathogenic Microbes ◽  
Antimicrobial Peptide Resistance ◽  
Significant Attention

: In recent years, over use of antibiotics has been raising its head to a serious problem all around the world as pathogens become drug resistant and create challenges to the medical field. This failure of most potent antibiotics that kill pathogens increases the thirst for research to look further way of killing pathogens. It has been led to the findings of antimicrobial peptide which is the most potent peptide to destroy pathogens. This review gives special emphasis to the usage of marine bacteria and other microorganisms for antimicrobial peptide (AMP) which are eco friendly as well as a developing class of natural and synthetic peptides with a wide spectrum of targets to pathogenic microbes. Consequently, a significant attention has been paid mainly to (i) the structure and types of anti microbial peptides and (ii) mode of action and mechanism of antimicrobial peptide resistance to pathogens. In addition to this, the designing of AMPs has been analysed thoroughly for reducing toxicity and developing better potent AMP. It has been done by the modified unnatural amino acids by amidation to target the control of biofilm and persister cell.

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.

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