Prolonged pre-incubation increases the susceptibility of <i>Galleria mellonella</i> larvae to bacterial and fungal infection

Browne, Niall; Surlis, Carla; Maher, Amie; Gallagher, Clair; Carolan, James C.; Clynes, Martin; Kavanagh, Kevin

doi:10.1080/21505594.2015.1021540

Cited by 29 publications

(45 citation statements)

References 43 publications

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“…Upon arrival the larvae were stored in the dark in wood shavings at 17°C and allowed to acclimatize for at least 2 days to control for the adverse physiological consequences of shipping [27]. G. mellonella larvae were used within 1 week due to the known physiological consequences of long-term storage of larvae [30]. Healthy G. mellonella larvae were selected by weight (175-225 mg), uniformity in color (little to no melanization), and responsiveness to touch.…”

Section: Methodsmentioning

confidence: 99%

The design and implementation of restraint devices for the injection of pathogenic microorganisms into Galleria mellonella

Fredericks

Roslund

Lee

et al. 2020

Preprint

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19 The injection of laboratory animals with pathogenic microorganisms poses a significant safety 20 risk because of the potential for injury by accidental needlestick. This is especially true for 21 researchers using invertebrate models of disease due to the small size of the animals and the 22 required precision and accuracy of the injection. The immobilization of the greater wax moth 23 larvae (Galleria mellonella) is often achieved by grasping a larva firmly between finger and 24 thumb. Needle resistant gloves or forceps can be used to reduce the risk of a needlestick but can 25 result in animal injury, a loss of throughput, and inconsistencies in experimental data.26 Immobilization devices are commonly used for the manipulation of small mammals, and in this 27 manuscript, we describe the construction of injection chambers that can be used to entrap and 28 restrain G. mellonella larvae prior to injection with pathogenic microbes. These devices 29 significantly reduce the manual handling of larvae and provide an engineering control to 30 protection against accidental needlestick injury, while maintaining a high rate of injection. 31 32 33 34 35 36 37 38 39 40 3 41 Introduction 42 The larvae of the greater wax moth Galleria mellonella is an important animal model for 43 studying host-pathogen interactions and for the discovery of novel antimicrobial therapeutics. 44 The popularity of this model organism is driven by the low cost of purchase and the reduced 45 ethical concerns for the experimental manipulation of insects. This allows the challenge of a 46 large number of larvae in a single experiment, which can improve the statistical power of an 47 assay. Starting in the 1940s, a diversity of viral, bacterial, fungal, and nematode pathogens, have 48 been studied for their ability to cause disease in G. mellonella larvae [1-15]. Importantly, G.49 mellonella can be maintained at mammalian body temperature and the outcomes of infection can 50 reproduce that of mammalian animal models [16][17][18]. This is likely due to similarities in the 51 innate immune response to pathogens mediated by elements of cellular and humoral immunity 52 between insects and mammals [19][20][21]. There are several methods to initiate an infection of a 53 larva with a pathogen, including topical application, feeding, oral gavage, and direct injection 54 into the hemocoel. The latter method is favored because of the ability to control the timing of 55 infection and the dosage of pathogen. 5657 Despite the benefits of G. mellonella larvae, there are challenges with using this animal model, 58 most notably standardizing the health and developmental stage of the larvae. This is especially 59 challenging when they are purchased from commercial sources that are primarily focused on 60 providing feed for the pet and angling communities [22,23], although there are efforts to develop 61 commercial pipelines for scientific-grade larvae [24]. Another difficulty is the manipulation and 62 restraint of small larvae during experimental injection of pat...

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Section: Methodsmentioning

confidence: 99%

The design and implementation of restraint devices for the injection of pathogenic microorganisms into Galleria mellonella

Fredericks

Roslund

Lee

et al. 2020

Preprint

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“…In these instances, immune priming offers short-term protection to larvae. Browne et al [53] found that when the larvae of G. mellonella were incubated at 30°C or 37°C for 72 h, they showed decreased survival compared to larvae exposed to the same conditions for a shorter period of time (24 h) prior to infection [53]. The density of hemocyte populations and the level of AMPs reached a peak at 24 h and then declined as the cost and maintenance of the primed response may outweigh its benefits.…”

Section: Thermal and Physical Stress As Immune Priming Agentsmentioning

confidence: 99%

Immune priming: the secret weapon of the insect world

2020

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Insects are a highly successful group of animals that inhabit almost every habitat and environment on Earth. Part of their success is due to a rapid and highly effective immune response that identifies, inactivates, and eliminates pathogens. Insects possess an immune system that shows many similarities to the innate immune system of vertebrates, but they do not possess an equivalent system to the antibody-mediated adaptive immune response of vertebrates. However, some insect do display a process known as immune priming in which prior exposure to a sublethal dose of a pathogen, or pathogen-derived material, leads to an elevation in the immune response rendering the insect resistant to a subsequent lethal infection a short time later. This process is mediated by an increase in the density of circulating hemocytes and increased production of antimicrobial peptides. Immune priming is an important survival strategy for certain insects while other insects that do not show this response may have colony-level behaviors that may serve to limit the success of pathogens. Insects are now widely used as in vivo models for studying microbial pathogens of humans and for assessing the in vivo efficacy of antimicrobial agents. Knowledge of the process of immune priming in insects is essential in these applications as it may operate and augment the perceived in vivo antimicrobial activity of novel compounds.

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“…9 In this edition of Virulence, the Kavanagh group report that larvae become increasingly susceptible to infection by pathogens as laboratory storage time increases, highlighting the need to consider this parameter when using the G. mellonella model. Browne et al 10 elaborate further in the study and relate this observation to a reduction in the total abundance of haemocytes that function in immune defense against pathogens and changes in the relative flux of metabolic pathways. Interestingly, the number of haemocytes after 3 weeks of incubation was approximately half that compared to the population at one week, while qualitative changes in the relative abundance of the various types of haemocytes were also reported.…”

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confidence: 92%

“…Interestingly, the number of haemocytes after 3 weeks of incubation was approximately half that compared to the population at one week, while qualitative changes in the relative abundance of the various types of haemocytes were also reported. 10 Both these factors probably contribute to reduced immune capacity and thus increased susceptibility to infection. The Browne et al study 10 and other related works raise awareness of ways to reduce inter-experimental variability and may help to standardise methods and permit more meaningful comparisons between certain types of experiment.…”

mentioning

confidence: 99%