DOCSLIB.ORG

Galleria Mellonella Infection Models for the Study of Bacterial Diseases and for Antimicrobial Drug Testing

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Galleria Mellonella Infection Models for the Study of Bacterial Diseases and for Antimicrobial Drug Testing VIRULENCE 2016, VOL. 7, NO. 3, 214–229 http://dx.doi.org/10.1080/21505594.2015.1135289 REVIEW Galleria mellonella infection models for the study of bacterial diseases and for antimicrobial drug testing Catherine Jia-Yun Tsaia,b, Jacelyn Mei San Loha,b, and Thomas Profta,b aDepartment of Molecular Medicine & Pathology, School of Medical Sciences, University of Auckland, Auckland, New Zealand; bMaurice Wilkins Center, University of Auckland, Auckland, New Zealand ABSTRACT ARTICLE HISTORY Galleria mellonella (greater wax moth or honeycomb moth) has been introduced as an alternative Received 18 November 2015 model to study microbial infections. G. mellonella larvae can be easily and inexpensively obtained in Revised 15 December 2015 large numbers and are simple to use as they don’t require special lab equipment. There are no Accepted 16 December 2015 ethical constraints and their short life cycle makes them ideal for large-scale studies. Although KEYWORDS insects lack an adaptive immune response, their innate immune response shows remarkable antimicrobial drug testing; similarities with the immune response in vertebrates. Galleria mellonella; Gram- This review gives a current update of what is known about the immune system of G. mellonella negative pathogens; Gram- and provides an extensive overview of how G. mellonella is used to study the virulence of Gram- positive pathogens; infection positive and Gram-negative bacteria. In addition, the use of G. mellonella to evaluate the efficacy of model; innate immunity; wax antimicrobial agents and experimental phage therapy are also discussed. The review concludes worm with a critical assessment of the current limitatons of G. mellonella infection models. Introduction One of the most commonly used models for studying use of G. mellonella does not require ethical approval microbial infections is the murine model. However, there and their short life span makes them ideal for high- are ethical, budgetary and logistical hurdles associated throughput studies. with the use of rodents as infection models. Firstly, With the completion of many microbial genome proj- maintaining a sufficient number of animals required to ects over the last decade, there is now a large number of obtain statistically relevant data is expensive and often so-called “hypothetical proteins” in the databases. These regarded as ethically objectionable. Secondly, mammals annotations are based on identified open reading frames, have lengthy reproduction times, which slow the prog- sometimes with predicted functions after bioinformatics ress of experimentation. More recently, Galleria mello- analyses, but often without an experimentally confirmed nella (greater wax moth or honeycomb moth) has been function. Many of these novel proteins are believed to be introduced as an alternative model to study microbial virulence factors and deciphering their functions will infections. More than 1000 articles have been published increase our understanding in disease mechanisms and on PubMed about the G. mellonella infection model, of ultimately provide a base for the development of novel which >200 were published in 2014–2015 alone demon- therapeutic agents. Unlike other invertebrate models strating the increasing popularity of this infection model. such as Caenorhabditis elegans and Drosophila mela- G. mellonella is an insect from the order Lepidoptera nogaster, G. mellonella larvae can survive at 37C and and the family Pyralidae (snout moths).1 It is in fact the therefore allow the investigation of temperature-depen- caterpillar larvae, or wax worm, and not the adult moth dent microbial virulence factors.3,4 that is used as an animal model. When compared with Insects have diverged from vertebrates approximately traditional mammalian model hosts, G. mellonella larvae 500 million years ago. Although vertebrates have devel- are cheaper to establish and easier to maintain, as they oped an adaptive immune response, their innate immune don’t require special lab equipment.2 Additionally, the response still retains remarkable similarities with the CONTACT Jacelyn Mei San Loh [email protected]; Thomas Proft [email protected] School of Medical Sciences, University of Auckland, Private Bag 92019, Auckland 1142, New Zealand. Color versions of one or more of the figures in this article can be found online at www.tandfonline.com/kvir. Supplemental data for this article can be accessed on the publisher’s website. © 2016 Catherine Jia-Yun Tsai, Jacelyn Mei San Loh, and Thomas Proft. Published with license by Taylor & Francis. This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted. VIRULENCE 215 immune response in insects.5 Over recent years, G. mel- characterized by a leaf-like shape and lysosomal enzymes lonella has been widely used as an infection model to within their cytoplasm. Granular cells are smaller and study bacterial and fungal infections and for assessing contain many granules in the cytoplasm. Encapsulation the efficacy of novel antimicrobial drugs. begins with the attachment of granular cells to the for- eign target triggering the release of material (e.g. plasma- tocyte spreading peptides, PSP) that promotes the The Galleria mellonella immune system attachment of multiple layers of plasmatocytes around 8,9 The innate immune response of insects consists of 2 the foreign target resulting in a smooth capsule. major parts, the cellular and the humoral immune Encapsulation is mainly associated with immune responses against larger microbes, such as protozoa and response. The cellular response is mediated by phago- 6 cytic cells, termed hemocytes. These are found within nematodes (including eggs and larvae). the hemolymph, which functions analogously to mam- Phagocytosis in insects and mammals is believed to be very similar and involves both plasmatocytes and malian blood. These cells are not only involved in phago- 7 cytosis, but also in encapsulation and clotting. The granular cells. G. mellonella expresses on hemocytes a humoral response is orchestrated by soluble effector protein with high homology to human calreticulin found on neutrophils, which is believed to be involved molecules that immobilize or kill the pathogen and 10 includes complement-like proteins, melanin, and anti- in non-self recognition in cellular defense reactions. microbial peptides (summarized in Table 1). Once phagocytosed, pathogens are killed by several mechanisms including reactive oxygen species (e.g., superoxide) generated by the oxidative burst, which is Cellular immune response initiated by the NADPH oxidase complex. In neutro- phils, the functional NADPH complex is formed after At least 8 types of hemocytes are found in insects, of translocation of proteins p47phox and p67phox from the fi which 6 different types have been identi ed in G. mello- cytosol to the plasma membrane. Homologous pro- 6 nella. These include prohemocytes, plasmatocytes, gran- teins p47 and p67 were identified in G. mellonella ular cells, coagulocytes, spherulocytes and oenocytoids. hemocytes and it was shown that superoxide produc- In G. mellonella, plasmatocytes and granular cells play a tion in both hemocytes and human neutrophils could key role in cellular defense and are involved in phagocy- be triggered by 12-myristate 13 acetate (PMA) and 7 tosis, nodule formation and encapsulation. Plasmato- inhibited by diphenyleneiodonium chloride.11,12 cytes are the most common hemocyte and are Table 1. Components of the G. mellonella innate immune Humoral immune response response. Opsonins a) cellular response G. mellonella produces several plasma proteins that serve hemocytes prohemocytes 5 as opsonins that recognize and bind to conserved micro- plasmatocytes 5,7 granular cells 5,7 bial components similar to pattern recognition receptors coagulocytes 5 in mammals. Apolipophorin-III (apoLp-III), a major 5 spherulocytes exchangeable lipid transport molecule plays a crucial oenocytoids 5,43 b) humoral response role in the innate immune response as a pattern recogni- opsonins apolipophorin-III (apoL-III) 13-15,18-20,121 tion molecule. ApoLp-III shows high affinity for hydro- peptidoglycan recognition proteins (PGRPs) 22 cationic protein 8 (GmCP8) 23 phobic ligands such as bacterial lipopolysaccharide (LPS) hemolin 25,26 and lipoteichoic acid (LTA).13,14 Furthermore, binding antimicrobial peptides b (AMPs) lysozyme 27,29,41,70 to -1, 3 glucan and to fungal conidia resulting in cecropin 27,30,61,89 increased cellular encapsulation was reported.15 ApoLp- moricin-like peptides 27,31 gloverin 22,27,34,89 III shows high homology with mammalian apolipopro- galiomycin 27,70 tein E (apoE), which is involved in LPS detoxification, gallerimycin 27,35,70 Galleria 27,28,33,41 the stimulation of phagocytosis and nitric oxide (NO) defensin 16,17 Gm proline-rich peptides 1 and 2 27,33,41 release from platelets. A multifunctional role has also Gm anionic peptide 1 and 2 27,29,33,41 been demonstrated for apoLp-III. For example, apoLp- inducible serine protease inhibitor 2 27 heliocin-like peptide 27 III stimulates increases in hemolymph antibacterial x-tox 27,36 activity and superoxide production by hemocytes18 and Gm apolipophoricin 33 melanization phenoloxidase pathway 37-39,122 enhances the activity
Load more

Recommended publications
  • Crystal Structure of the Vibrio Cholerae Cytolysin Heptamer Reveals Common Features Among Disparate Pore-Forming Toxins
    Crystal structure of the Vibrio cholerae cytolysin heptamer reveals common features among disparate pore-forming toxins Swastik De and Rich Olson1 Department of Molecular Biology and Biochemistry, Wesleyan University, 52 Lawn Avenue, Middletown, CT 06459 Edited* by Pamela J. Bjorkman, California Institute of Technology, Pasadena, CA, and approved March 21, 2011 (received for review November 19, 2010) Pore-forming toxins (PFTs) are potent cytolytic agents secreted are the staphylococcal PFTs, which display weak sequence iden- by pathogenic bacteria that protect microbes against the cell- tity (approximately 15%) but strong structural similarity with mediated immune system (by targeting phagocytic cells), disrupt the VCC core domain (6). Structures exist for the water-soluble epithelial barriers, and liberate materials necessary to sustain bicomponent LukF (7, 8) and LukS (9) toxins as well as the growth and colonization. Produced by gram-positive and gram- homomeric α-hemolysin (α-HL) heptamer (10), which represents negative bacteria alike, PFTs are released as water-soluble mono- the only fully assembled β-PFTcrystal structure determined until meric or dimeric species, bind specifically to target membranes, and now. Distinct in sequence and structure from VCC (and each assemble transmembrane channels leading to cell damage and/or other), but with a similar heptameric stoichiometry, are the ex- lysis. Structural and biophysical analyses of individual steps in tensively studied aerolysin (11) and anthrax toxins (12) produced the assembly
    [Show full text]
  • The Role of Streptococcal and Staphylococcal Exotoxins and Proteases in Human Necrotizing Soft Tissue Infections
    toxins Review The Role of Streptococcal and Staphylococcal Exotoxins and Proteases in Human Necrotizing Soft Tissue Infections Patience Shumba 1, Srikanth Mairpady Shambat 2 and Nikolai Siemens 1,* 1 Center for Functional Genomics of Microbes, Department of Molecular Genetics and Infection Biology, University of Greifswald, D-17489 Greifswald, Germany; [email protected] 2 Division of Infectious Diseases and Hospital Epidemiology, University Hospital Zurich, University of Zurich, CH-8091 Zurich, Switzerland; [email protected] * Correspondence: [email protected]; Tel.: +49-3834-420-5711 Received: 20 May 2019; Accepted: 10 June 2019; Published: 11 June 2019 Abstract: Necrotizing soft tissue infections (NSTIs) are critical clinical conditions characterized by extensive necrosis of any layer of the soft tissue and systemic toxicity. Group A streptococci (GAS) and Staphylococcus aureus are two major pathogens associated with monomicrobial NSTIs. In the tissue environment, both Gram-positive bacteria secrete a variety of molecules, including pore-forming exotoxins, superantigens, and proteases with cytolytic and immunomodulatory functions. The present review summarizes the current knowledge about streptococcal and staphylococcal toxins in NSTIs with a special focus on their contribution to disease progression, tissue pathology, and immune evasion strategies. Keywords: Streptococcus pyogenes; group A streptococcus; Staphylococcus aureus; skin infections; necrotizing soft tissue infections; pore-forming toxins; superantigens; immunomodulatory proteases; immune responses Key Contribution: Group A streptococcal and Staphylococcus aureus toxins manipulate host physiological and immunological responses to promote disease severity and progression. 1. Introduction Necrotizing soft tissue infections (NSTIs) are rare and represent a more severe rapidly progressing form of soft tissue infections that account for significant morbidity and mortality [1].
    [Show full text]
  • Directed Antigen Delivery As a Vaccine Strategy for an Intracellular Bacterial Pathogen
    Directed antigen delivery as a vaccine strategy for an intracellular bacterial pathogen H. G. Archie Bouwer*, Christine Alberti-Segui†, Megan J. Montfort*, Nathan D. Berkowitz†, and Darren E. Higgins†‡ *Immunology Research, Earle A. Chiles Research Institute and Veterans Affairs Medical Center, Portland, OR 97239; and †Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115 Edited by John J. Mekalanos, Harvard Medical School, Boston, MA, and approved February 8, 2006 (received for review October 27, 2005) We have developed a vaccine strategy for generating an attenu- murine infection model. Many facets of pathogenesis and protective ated strain of an intracellular bacterial pathogen that, after uptake immunity to Lm are well characterized. After uptake by APCs, Lm by professional antigen-presenting cells, does not replicate intra- escapes from the phagocytic vacuole into the cytosol, an event cellularly and is readily killed. However, after degradation of the facilitated by a secreted pore-forming cytolysin, listeriolysin O vaccine strain within the phagolysosome, target antigens are (LLO) (8). After escape from the vacuole, Lm replicates freely released into the cytosol for endogenous processing and presen- within the cytosol, allowing bacterial products to access the endog- tation for stimulation of CD8؉ effector T cells. Applying this enous MHC Class I presentation pathway. Consistent with this strategy to the model intracellular pathogen Listeria monocyto- cytosolic replication niche, protective antilisterial immunity de- genes, we show that an intracellular replication-deficient vaccine pends on Lm-specific CD8ϩ effector T cells, with antibody playing strain is cleared rapidly in normal and immunocompromised ani- no significant role (9).
    [Show full text]
  • From Phagosome Into the Cytoplasm on Cytolysin, Listeriolysin O, After Evasion Listeria Monocytogenes in Macrophages by Dependen
    Dependency of Caspase-1 Activation Induced in Macrophages by Listeria monocytogenes on Cytolysin, Listeriolysin O, after Evasion from Phagosome into the Cytoplasm This information is current as of September 23, 2021. Hideki Hara, Kohsuke Tsuchiya, Takamasa Nomura, Ikuo Kawamura, Shereen Shoma and Masao Mitsuyama J Immunol 2008; 180:7859-7868; ; doi: 10.4049/jimmunol.180.12.7859 http://www.jimmunol.org/content/180/12/7859 Downloaded from References This article cites 50 articles, 25 of which you can access for free at: http://www.jimmunol.org/content/180/12/7859.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 23, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2008 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Dependency of Caspase-1 Activation Induced in Macrophages by Listeria monocytogenes on Cytolysin, Listeriolysin O, after Evasion from Phagosome into the Cytoplasm1 Hideki Hara, Kohsuke Tsuchiya, Takamasa Nomura, Ikuo Kawamura, Shereen Shoma, and Masao Mitsuyama2 Listeriolysin O (LLO), an hly-encoded cytolysin from Listeria monocytogenes, plays an essential role in the entry of this pathogen into the macrophage cytoplasm and is also a key factor in inducing the production of IFN-␥ during the innate immune stage of infection.
    [Show full text]
  • EFFECT of STORAGE DURATION on the STORED PUPAE of PARASITOID Bracon Hebetor (Say) and ITS IMPACT on PARASITOID QUALITY M
    ISSN 0258-7122 (Print), 2408-8293 (Online) Bangladesh J. Agril. Res. 41(2): 297-310, June 2016 EFFECT OF STORAGE DURATION ON THE STORED PUPAE OF PARASITOID Bracon hebetor (Say) AND ITS IMPACT ON PARASITOID QUALITY M. S. ALAM1, M. Z. ALAM2, S. N. ALAM3 M. R. U. MIAH4 AND M. I. H. MIAN5 Abstract The ecto-endo larval parasitoid, Bracon hebetor (Say) is an important bio- control agent. Effective storage methods for B. hebetor are essential for raising its success as a commercial bio-control agent against lepidopteran pests. The study was undertaken to determine the effect of storage duration on the pupae of Bracon hebetor in terms of pupal survival, adult emergence, percent parasitism, female and male longevity, female fecundity and sex ratio. Three to four days old pupae were stored for 0, 1, 2, 3, 4, 5, 6, 7 and 8 weeks at 4 ± 1oC. The ranges of time for adult emergence from stored pupae, production of total adult, survivability of pupae, parasitism of host larvae by the parasitoid, longevity of adult female and male and fecundity were 63.0 -7.5 days, 6.8-43.8/50 host larvae, 13.0-99.5%, 0.0 -97.5%, 0.00-20.75 days, 0.00-17.25 days and 0.00- 73.00/50 female, respectively. The time of adult emergence and mortality of pupae increased but total number of adult emergence, survivability of pupae, longevity of adult female and male decreased gradually with the progress of storage period of B. hebetor pupae. The prevalence of male was always higher than that of female.
    [Show full text]
  • Penetration of Stratified Mucosa Cytolysins Augment Superantigen
    Cytolysins Augment Superantigen Penetration of Stratified Mucosa Amanda J. Brosnahan, Mary J. Mantz, Christopher A. Squier, Marnie L. Peterson and Patrick M. Schlievert This information is current as of September 25, 2021. J Immunol 2009; 182:2364-2373; ; doi: 10.4049/jimmunol.0803283 http://www.jimmunol.org/content/182/4/2364 Downloaded from References This article cites 76 articles, 24 of which you can access for free at: http://www.jimmunol.org/content/182/4/2364.full#ref-list-1 Why The JI? Submit online. http://www.jimmunol.org/ • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average by guest on September 25, 2021 Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2009 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Cytolysins Augment Superantigen Penetration of Stratified Mucosa1 Amanda J. Brosnahan,* Mary J. Mantz,† Christopher A. Squier,† Marnie L. Peterson,‡ and Patrick M. Schlievert2* Staphylococcus aureus and Streptococcus pyogenes colonize mucosal surfaces of the human body to cause disease. A group of virulence factors known as superantigens are produced by both of these organisms that allows them to cause serious diseases from the vaginal (staphylococci) or oral mucosa (streptococci) of the body.
    [Show full text]
  • Antibodies Against Pneumolysine and Their Applications
    Europaisches Patentamt J European Patent Office © Publication number: 0 687 688 A1 Office europeen des brevets EUROPEAN PATENT APPLICATION published in accordance with Art. 158(3) EPC © Application number: 95903822.5 © int. ci.<>: C07K 16/12, C07K 5/10, C07K 14/315, C07K 14/33, @ Date of filing: 17.12.94 A61 K 39/395, G01 N 33/569 © International application number: PCT/ES94/00135 © International publication number: WO 95/16711 (22.06.95 95/26) © Priority: 17.12.93 ES 9302615 Inventor: MENDEZ, Francisco J. 14.02.94 ES 9400265 Fernando Vela, 20 13.08.94 GB 9416393 E-33011 Oviedo (ES) Inventor: DE LOS TOYOS, Juan R. © Date of publication of application: Marcelino Suarez, 13 20.12.95 Bulletin 95/51 E-33012 Oviedo (ES) Inventor: VAZQUEZ, Fernando @ Designated Contracting States: Cervantes, 33 AT BE CH DE DK ES FR GB IE IT LI NL SE E-33005 Oviedo (ES) Inventor: MITCHELL, Timmothy © Applicant: UNIVERSIDAD DE OVIEDO 25 Mawbys Lane, Calle San Francisco, 3 Appleby Magna E-33003 Oviedo (ES) Swadlincote DE12 7AA (GB) Applicant: UNIVERSITY OF LEICESTER Inventor: ANDREW, Peter University Road 7 Lane Leicester, LE1 7RH (GB) Chapel Leicester LE2 3WF (GB) © Inventor: FLEITES, Ana Inventor: MORGAN, Peter J. Campoamor, 28 40 Medina Drive, E-33001 Oviedo (ES) Tollerton Inventor: FERNANDEZ, Jes s M. Nottingham NG12 4EP (GB) Marques de Lozoya, 12 E-28007 Madrid (ES) Inventor: HARDISSON, Carlos © Representative: Ibanez, Jose Francisco Arguelles, 19 Rodriguez San Pedro, 10 E-33002 Oviedo (ES) E-28015 Madrid (ES) 00 00 CO (54) ANTIBODIES AGAINST PNEUMOLYSINE AND THEIR APPLICATIONS IV 00 CO © An object of the invention is the generation of hybridomas and the production and characterization of the corresponding monoclonal antibodies specific to pneumolysine, which could be used for various purposes, including diagnosis, prophylaxis and therapy.
    [Show full text]
  • Wax Worms (Galleria Mellonella) As Potential Bioremediators for Plastic Pollution Student Researcher: Alexandria Elliott Faculty Mentor: Danielle Garneau, Ph.D
    Wax Worms (Galleria mellonella) as Potential Bioremediators for Plastic Pollution Student Researcher: Alexandria Elliott Faculty Mentor: Danielle Garneau, Ph.D. Center for Earth and Environmental Science SUNY Plattsburgh, Plattsburgh, NY 12901 Plastic Pollution Life History Stages Results Discussion • 30 million tons of plastic Combination of holes in plastic and • Egg stage: average length (0.478mm) • waste is generated annually and width (0.394mm) and persists 3-10 nylon in frass suggests worms are in the USA (Coalition 2018). days (Kwadha et al. 2017)(Fig. 3). digesting plastic (Figs. 6,7). • Larval stage: max length (30mm), white Of the plastic pilot trials which • 50% landfill, < 10% cream in color, possess 3 apical teeth, exhibited signs of feeding, two were HDPE (Fig. 6). recycled (PlasticsEurope, and persists 22-69 days. A Plastics The Facts 2013) • Pre-pupal/Pupal stage: length (12- • Bombelli et al. (2017) and Yang et al. 20mm) and persists 3-12 and 8-10 days, (2014) found wax worms were capable respectively. All extremities are glued to of PE consumption. • 10% of world’s plastic waste body with molting substance. Common bond (CH -CH ) in PE is 2 2 ends up in ocean 70% • Moth stage: sexual dimorphism is same as that in beeswax (Bombelli et sinks 30% floats in Fig. 1. Plastic Use distinct. Moths max length (20mm) and Fig. 5. Change in worm weight as a function of plastic pilot trial. al. 2017). Fig. 9. PE degradation as currents (Gyres, Fig. 2). (Plastics Europe). persists on average 6-14 days (males) B • Greater negative change in worm weight (g)/day for all FT-IR shows degradation of PE (i.e., evidenced by FT-IR PE and 23 days (females).
    [Show full text]
  • Purification and Characterization of an Extracellular Cytolysin Produced by Vibrio Damsela MAHENDRA H
    INFECTION AND IMMUNITY, JUlY 1985, P. 25-31 Vol. 49, No. 1 0019-9567/85/070025-07$02.00/0 Copyright © 1985, American Society for Microbiology Purification and Characterization of an Extracellular Cytolysin Produced by Vibrio damsela MAHENDRA H. KOTHARY AND ARNOLD S. KREGER* Department of Microbiology and Immunology, The Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, North Carolina 27103 Received 10 January 1985/Accepted 26 March 1985 Large amounts of an extremely potent extracellular cytolysin produced by the halophilic bacterium Vibrio damsela were obtained free of detectable contamination with medium constituents and other bacterial products by sequential ammonium sulfate precipitation, gel filtration with Sephadex G-100, and hydrophobic interaction chromatography with phenyl-Sepharose CL-4B. The cytolysin is heat labile and protease sensitive and has a molecular weight (estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis) of ca. 69,000 and an isoelectric point of ca. 5.6. The first 10 amino-terminal amino acid residues of the cytolysin are Phe-Thr-Gln- Trp-Gly-Gly-Ser-Gly-Leu-Thr. The cytolysin was very active against erythrocytes from 4 of the 18 animal species examined (mice, rats, rabbits, damselfish) and against Chinese hamster ovary cells and was lethal for mice (ca. 1 ,Ig/kg, intraperitoneal median lethal dose). Lysis of mouse erythrocytes by the cytolysin is a multi-hit, at least two-step process consisting of a temperature-independent, toxin-binding step followed by a temperature-dependent, membrane-perturbation step(s). The halophilic bacterium Vibrio damsela, one of seven The purified cytolysin preparation (stage 4) was assayed new Vibrio species recognized since 1976 as possible causes for activity against Chinese hamster ovary cells as described of human disease (26, 30), is an opportunistic pathogen that by Kreger and Lockwood (17).
    [Show full text]
  • Effect of Gamma Radiation and Entomopathogenic Nematodes on Greater Wax Moth, Galleria Mellonella (Linnaeus) [Lep., Pyralidae]
    Effect of gamma radiation and entomopathogenic nematodes on greater wax moth, Galleria mellonella (Linnaeus) [Lep., Pyralidae] A Thesis Submitted in Partial Fulfillment of the Requirements for the Degree of M. Sc. in Entomology By Rehab Mahmoud Sayed Ali B.Sc. Entomology, 2001 Supervised by Prof. Dr. Prof. Dr. Mohamed Adel Hussein Soryia El-Tantawy Hafez Professor of Entomology Professor of Entomology Entomology Department, Faculty of Entomology Department, Faculty of Science, Ain Shams University Science, Ain Shams University Prof. Dr . Hedayat-Allah Mahmoud Mohamed Salem Professor of Entomology Natural Products Department, National Center for Radiation Sciences & Technology Department of Entomology Faculty of Science Ain Shams University 2008 Faculty of Science Approval Sheet M. Sc. Thesis Name: Rehab Mahmoud Sayed Ali Title: Effect of gamma radiation and entomopathogenic nematodes on the greater wax moth, Galleria mellonella (Linnaeus) [Lep., Pyralidae] This Thesis for M. Sc. Degree in Entomology has been approved by: Prof. Dr. Hedayat-Allah Mahmoud Mohamed Salem Professor of Entomology, Natural Products Department, National Center for Radiation Sciences & Technology. Prof. Dr. Mohamed Adel Hussein Professor of Entomology, Entomology Department, Faculty of Science, Ain Shams University. Prof. Dr. Mohamed Salem Abd EL-Wahed Professor of Entomology, Faculty of Agriculture, Ain Shams University. Prof. Dr . Mahmoud Mohamed Saleh Professor of Nematodes, Pests and Plant Protection Department, National research centre. Date: / / Ain Shams
    [Show full text]
  • Dodecanol, Metabolite of Entomopathogenic Fungus
    www.nature.com/scientificreports OPEN Dodecanol, metabolite of entomopathogenic fungus Conidiobolus coronatus, afects fatty acid composition and cellular immunity of Galleria mellonella and Calliphora vicina Michalina Kazek1*, Agata Kaczmarek1, Anna Katarzyna Wrońska1 & Mieczysława Irena Boguś1,2 One group of promising pest control agents are the entomopathogenic fungi; one such example is Conidiobolus coronatus, which produces a range of metabolites. Our present fndings reveal for the frst time that C. coronatus also produces dodecanol, a compound widely used to make surfactants and pharmaceuticals, and enhance favors in food. The main aim of the study was to determine the infuence of dodecanol on insect defense systems, i.e. cuticular lipid composition and the condition of insect immunocompetent cells; hence, its efect was examined in detail on two species difering in susceptibility to fungal infection: Galleria mellonella and Calliphora vicina. Dodecanol treatment elicited signifcant quantitative and qualitative diferences in cuticular free fatty acid (FFA) profles between the species, based on gas chromatography analysis with mass spectrometry (GC/MS), and had a negative efect on G. mellonella and C. vicina hemocytes and a Sf9 cell line in vitro: after 48 h, almost all the cells were completely disintegrated. The metabolite had a negative efect on the insect defense system, suggesting that it could play an important role during C. coronatus infection. Its high insecticidal activity and lack of toxicity towards vertebrates suggest it could be an efective insecticide. In their natural environment, insects have to cope with a variety of microorganisms, and as such, have developed a complex and efcient defense system. Te frst line of defense is a cuticle formed of several layers, with an epi- cuticle on the outside, a procuticle underneath it and an epidermis beneath that.
    [Show full text]
  • Plodia Interpunctella H
    ﺗﺤﻘﻴﻘﺎت آﻓﺎت ﮔﻴﺎﻫﻲ 1397 -8 (1): 69-82 ﺑﻬﻴﻨﻪ ﺳﺎزي ﻣﺼﺮف ﺣﺸﺮه ﻛﺶ ﻛﻠﺮﭘﺎﻳﺮﻳﻔﻮس ﻋﻠﻴﻪ ﺷﺐ ﭘﺮه ﻫﻨﺪي Plodia interpunctella ، ﺷﺐ ﭘﺮه ﻣﺪﻳﺘﺮاﻧﻪ اي آرد Anagasta kuehniella و ﺷﺐ ﭘﺮه ﻣﻮم ﺧﻮار ﺑﺰرگ Galleria mellonella ﺑﺎ روش ﺳﻄﺢ ﭘﺎﺳﺦ در ﺷﺮاﻳﻂ آزﻣﺎﻳﺸﮕﺎﻫﻲ ﻋﻠﻴﺮﺿﺎ ﺷﻌﺒﺎﻧﻲ ﻧﮋاد1 و ﻣﺮﻳﻢ ﻋﺠﻢ ﺣﺴﻨﻲ1* 1 - داﻧﺸﻜﺪه ﻛﺸﺎورزي، داﻧﺸﮕﺎه ﺻﻨﻌﺘﻲ ﺷﺎﻫﺮود، ﺷﺎﻫﺮود ( ﺗﺎرﻳﺦ درﻳﺎﻓﺖ /8: 11/ 96 ﺗﺎرﻳﺦ ﭘﺬﻳﺮش : 97/3/30) ﭼﻜﻴﺪه ﺷﺐ ﭘﺮه ﻫﻨﺪي .Plodia interpunctella H، ﺷﺐ ﭘﺮه ﻣﺪﻳﺘﺮاﻧﻪ اي آرد .Anagasta kuehniella Z و ﺷﺐ ﭘﺮه ﻣﻮم ﺧﻮار ﺑﺰرگ Galleria .mellonella L ا ز آﻓﺎت ﺑﺴﻴﺎر ﻣﻬﻢ ﻣﺤﺼﻮﻻت اﻧﺒﺎري ﻫﺴﺘﻨﺪ ﻛﻪ ﺑﻪ ﻣﻨﻈﻮر ﻛﻨﺘﺮل آنﻫﺎ، ﺑﻪ ﻃﻮر ﻣﻌﻤﻮل از ﺣﺸﺮه ﻛﺶﻫﺎي ﺷﻴﻤﻴﺎﻳﻲ اﺳﺘﻔﺎده ﻣﻲ - ﺷﻮد. اﺳﺘﻔﺎده ﺑﻲ روﻳﻪ از ﺣﺸﺮه ﻛﺶﻫﺎ، ﻣﺸﻜﻼت ﺟﺪي زﻳﺴﺖ ﻣﺤﻴﻄﻲ و ﻫﻤﭽﻨﻴﻦ ﻋﻮارض ﻣﺰﻣﻨﻲ روي ﺳﻼﻣﺖ اﻧﺴﺎن اﻳﺠﺎد ﻣﻲ ﻛﻨﺪ، ﺑﻨﺎﺑﺮاﻳﻦ ﺑﺎ اﻧﺘﺨﺎب اﻟﮕﻮي ﺻﺤﻴﺢ و ﺑﻪ ﻛﺎ رﮔﻴﺮي روش ﻫﺎي ﻣﻨﺎﺳﺐ ﻣﻲ ﺗﻮان ﻣﻴﺰان ﻣﺼﺮف ﺣﺸﺮه ﻛﺶ را ﺑﻬﻴﻨﻪ ﻛﺮد. در اﻳﻦ ﭘﮋوﻫﺶ از روش ﺳﻄﺢ ﭘﺎﺳﺦ ﺑﻪ ﻣﻨﻈﻮر ﺗﻌﻴﻴﻦ ﻧﻘﺎط ﺑﻬﻴﻨﻪ ﻣﺼﺮف ﺣﺸﺮه ﻛﺶ ﻛﻠﺮﭘﺎﻳﺮﻳﻔﻮس، ﺟﻬﺖ دﺳﺘﻴﺎﺑﻲ ﺑﻪ ﺑﻴﺸﻴﻨﻪ ﻣﺮگ و ﻣﻴﺮ ﺑﺎ ﻛﻤﺘﺮ ﻳﻦ دز ﻣﺼﺮﻓﻲ اﺳﺘﻔﺎده ﺷﺪ و ﺗﺎﺛﻴﺮ دﻣﺎ ( 25-35 درﺟﻪ ﺳﻠﺴﻴﻮس)، رﻃﻮﺑﺖ ( 60-80 %) و د ز ﻣﺼﺮف ( 1400-2000 ﻣﻴﻜﺮوﻟﻴﺘﺮ) ﺑﺮ ﻣﻴﺰان ﻣﺮگ و ﻣﻴﺮ ﻻرو ﺳﻦ ﭘﻨﺠﻢ ﺳﻪ آﻓﺖ ﻣﻬﻢ اﻧﺒﺎري ﻣﻮرد ارزﻳﺎﺑﻲ ﻗﺮار ﮔﺮﻓﺖ. آزﻣﺎﻳﺶ ﻫﺎ ﺑﺮ اﺳﺎس ﻃﺮح ﻣﺮﻛﺐ ﻣﺮﻛﺰي، اﻧﺠﺎم ﺷﺪ. ﺑﻴﺸﻴﻨﻪ ﻣﺮگ و ﻣﻴﺮ ﺑﺎ اﻳﻦ روش، 9/7 ﻋﺪد ﻻرو ﺳﻦ ﭘﻨﺞ ﺷﺐ ﭘﺮه ﻫﻨﺪي در ﺷﺮاﻳﻂ دﻣﺎﻳ ﻲ 35 درﺟﻪ ﺳﻠﺴﻴﻮس، رﻃﻮﺑﺖ 80 % و دز 1400 ﻣﻴﻜﺮوﻟﻴﺘﺮ ﺣﺸﺮه ﻛﺶ در 1000 ﻣﻴﻠﻲ ﻟﻴﺘﺮ آب، 1/7 ﻋﺪد ﻻرو ﺷﺐ ﭘﺮه ﻣﺪﻳﺘﺮاﻧﻪ اي در دﻣﺎي 35 درﺟﻪ ﺳﻠﺴﻴﻮس، رﻃﻮﺑﺖ 75 % و دز 1400 ﻣﻴﻜﺮوﻟﻴﺘﺮ ﺣﺸﺮه ﻛﺶ در 1000 ﻣﻴﻠﻲ ﻟﻴﺘﺮ آب و ﺑﺮاي ﺷﺐ ﭘﺮه ﻣﻮم ﺧﻮار ﺑﺰرگ 7/9 ﻋﺪد ﻻرو ﺳﻦ ﭘﻨﺞ در دﻣﺎي 35 درﺟﻪ ﺳﻠﺴﻴﻮس، رﻃﻮﺑﺖ 80 % و دز ﻣﺼﺮﻓﻲ 1500 ﻣﻴﻜﺮوﻟﻴﺘﺮ ﺣﺸﺮه ﻛﺶ در 1000 ﻣﻴﻠﻲ ﻟﻴﺘﺮ ﺗﻌﻴﻴﻦ ﺷﺪ .
    [Show full text]