scholarly journals Bacillus thuringiensis Spores and Vegetative Bacteria: Infection Capacity and Role of the Virulence Regulon PlcR Following Intrahaemocoel Injection of Galleria mellonella

Insects ◽  
2019 ◽  
Vol 10 (5) ◽  
pp. 129 ◽  
Author(s):  
Christophe Buisson ◽  
Michel Gohar ◽  
Eugénie Huillet ◽  
Christina Nielsen-LeRoux
Keyword(s):  
Bacillus Thuringiensis ◽  
Galleria Mellonella ◽  
Lethal Dose ◽  
Intestinal Barrier ◽  
Oral Route ◽  
Infection Model ◽  
Wild Type ◽  
Response Curves ◽  
Vegetative Cells ◽  
Crystal Toxins

Bacillus thuringiensis is an invertebrate pathogen that produces insecticidal crystal toxins acting on the intestinal barrier. In the Galleria mellonella larvae infection model, toxins from the PlcR virulence regulon contribute to pathogenicity by the oral route. While B. thuringiensis is principally an oral pathogen, bacteria may also reach the insect haemocoel following injury of the cuticle. Here, we address the question of spore virulence as compared to vegetative cells when the wild-type Bt407cry- strain and its isogenic ∆plcR mutant are inoculated directly into G. mellonella haemocoel. Mortality dose-response curves were constructed at 25 and 37 °C using spores or vegetative cell inocula, and the 50% lethal dose (LD50) in all infection conditions was determined after 48 h of infection. Our findings show that (i) the LD50 is lower for spores than for vegetative cells for both strains, while the temperature has no significant influence, and (ii) the ∆plcR mutant is four to six times less virulent than the wild-type strain in all infection conditions. Our results suggest that the environmental resistant spores are the most infecting form in haemocoel and that the PlcR virulence regulon plays an important role in toxicity when reaching the haemocoel from the cuticle and not only following ingestion.

scholarly journals Pathogenicity and Immunogenicity of a Mutant Strain ofListeria monocytogenesin the Chicken Infection Model

2011 ◽  
Vol 18 (3) ◽  
pp. 500-505 ◽  
Author(s):  
Yuelan Yin ◽  
Debin Tian ◽  
Hongmei Jiao ◽  
Chenju Zhang ◽  
Zhiming Pan ◽  
...  
Keyword(s):  
Immune Response ◽  
Mutant Strain ◽  
Parent Strain ◽  
Lethal Dose ◽  
Oral Route ◽  
Infection Model ◽  
Listeriolysin O ◽  
Wild Type ◽  
Strong Immune Response ◽  
Booster Immunization

ABSTRACTListeria monocytogeneshas been exploited as a vaccine carrier based upon its ability to induce a strong cell-mediated immune response. At present, the safety of live, attenuatedL. monocytogenesvaccines in patients is being studied in clinical trials.L. monocytogenesis also an attractive vaccine vector for use in poultry; however, the pathogenicity and immunogenicity of this organism in poultry remain to be fully elucidated. In this study, we investigated the pathogenicity and immunogenicity of anactA- andplcB-deficientL. monocytogenesstrain, yzuLM4ΔactA/plcB, and its wild-type parent strain, yzuLM4, in an avian infection model. The results showed that the wild-type strain could infect ISA brown chickens, causing serious tissue disruptions, including various degrees of degeneration, necrotic lesions, and inflammatory cell infiltration in the liver, spleen, heart, and kidney. However, the mutant strain showed reduced virulence in embryonated eggs compared with that of the parent strain (the 50% lethal dose [LD50] was 3 logs higher). The mutant strain also showed low virulence in chickens and was rapidly eliminated by the host. There were no obvious pathological changes in tissue sections, but the mutant strain still retained the ability to stimulate high levels of antibody against the protein listeriolysin O (LLO). Booster immunization with the mutant strain led to rapid bacterial clearance from the livers and spleens of chickens challenged by the intramuscular route or the oral route. Collectively, our data suggest that the wild-type serotype 1/2aL. monocytogenesstrain can cause serious disease in chickens but the mutant strain with a deletion of theactAandplcBgenes is less virulent but induces a strong immune response. This mutant strain ofL. monocytogenesis therefore a promising candidate as a safe and effective vector for the delivery of heterologous antigens to prevent zoonosis and infectious disease in poultry.

scholarly journals Pneumococcal Surface Protein C Contributes to Sepsis Caused by Streptococcus pneumoniae in Mice

2004 ◽  
Vol 72 (5) ◽  
pp. 3077-3080 ◽  
Author(s):  
Francesco Iannelli ◽  
Damiana Chiavolini ◽  
Susanna Ricci ◽  
Marco Rinaldo Oggioni ◽  
Gianni Pozzi
Keyword(s):  
Streptococcus Pneumoniae ◽  
Protein C ◽  
Lethal Dose ◽  
Surface Protein ◽  
Infection Model ◽  
Wild Type ◽  
Mutant Strains ◽  
Type 3

ABSTRACT The role of pneumococcal surface protein C (PspC; also called SpsA, CbpA, and Hic) in sepsis by Streptococcus pneumoniae was investigated in a murine infection model. The pspC gene was deleted in strains D39 (type 2) and A66 (type 3), and the mutants were tested by being injected intravenously into mice. The animals infected with the mutant strains showed a significant increase in survival, with the 50% lethal dose up to 250-fold higher than that for the wild type. Our findings indicate that PspC affords a decisive contribution to sepsis development.

EFFECT OF TEMPERATURE ON THE PATHOGENESIS OF BACILLUS THURINGIENSIS BERLINER IN LARVAE OF THE SPRUCE BUDWORM, CHORISTONEURA FUMIFERANA CLEM. (LEPIDOPTERA: TORTRICIDAE)

1994 ◽  
Vol 126 (4) ◽  
pp. 1061-1065 ◽  
Author(s):  
Kees van Frankenhuyzen
Keyword(s):  
Bacillus Thuringiensis ◽  
Bacterial Growth ◽  
Choristoneura Fumiferana ◽  
Larval Mortality ◽  
Lethal Dose ◽  
Spruce Budworm ◽  
Effect Of Temperature ◽  
Vegetative Cells ◽  
Time Required ◽  
Peak Abundance

AbstractThe relationship between temperature and pathogenesis of Bacillus thuringiensis Berliner var. kurstaki in infected larvae of the eastern spruce budworm, Choristoneura fumiferana Clem., was investigated to determine if more rapid death of larvae with an increase in temperature could be accounted for by enhanced bacterial growth. Cumulative mortality of larvae force-fed with a lethal dose of HD-1-S-1980 peaked within 2 days at 25 °C, 3 days at 19 °C, 7 days at 16 °C, and 21 days at 13 °C. The progress of bacterial growth in the larvae was followed from spore germination to cell lysis, and was completed within 4 days at 25 °C, 6 days at 22 °C, 12 days at 19 °C, 14 days at 16 °C, and > 28 days at 13 °C. Peak abundance of vegetative cells in the larvae was observed after 1 day at 25 °C, 2 days at 22 °C, 3 days at 19 °C, 7 days at 16 °C, and 21 days at 13 °C, and thus coincided almost exactly with the time required for maximum larval mortality. This correlation suggests that the observed effect of temperature on progression of larval mortality was due to its effect on the proliferation of vegetative cells in the infected larvae, and that bacterial septicemia makes an important contribution to death.

scholarly journals ThemtfATranscription Factor Gene Controls Morphogenesis, Gliotoxin Production, and Virulence in the Opportunistic Human Pathogen Aspergillus fumigatus

2014 ◽  
Vol 13 (6) ◽  
pp. 766-775 ◽  
Author(s):  
Timothy D. Smith ◽  
Ana M. Calvo
Keyword(s):  
Aspergillus Fumigatus ◽  
Protease Activity ◽  
Galleria Mellonella ◽  
Molecular Genetic ◽  
Colony Growth ◽  
Infection Model ◽  
Slight Reduction ◽  
Wild Type ◽  
Content Type ◽  
Factor Gene

ABSTRACTAspergillus fumigatusis the leading causative agent of invasive aspergillosis (IA). The number of cases is on the rise, with mortality rates as high as 90% among immunocompromised patients. Molecular genetic studies inA. fumigatuscould provide novel targets to potentially set the basis for antifungal therapies. In the current study, we investigated the role of the transcription factor genemtfAinA. fumigatus. Our results revealed thatmtfAplays a role in the growth and development of the fungus. Deletion or overexpression ofmtfAleads to a slight reduction in colony growth, as well as a reduction in conidiation levels, in the overexpression strain compared to the wild-type strain. Furthermore, production of the secondary metabolite gliotoxin increased whenmtfAwas overexpressed, coinciding with an increase in the transcription levels of the gliotoxin genesgliZandgliPwith respect to the wild type. In addition, our study showed thatmtfAis also necessary for normal protease activity inA. fumigatus; deletion ofmtfAresulted in a reduction of protease activity compared to wild-type levels. Importantly, the absence ofmtfAcaused a decrease in virulence in theGalleria mellonellainfection model, indicating thatmtfAis necessary forA. fumigatuswild-type pathogenesis.

scholarly journals SlyA Is a Transcriptional Regulator Involved in the Virulence of Enterococcus faecalis

2011 ◽  
Vol 79 (7) ◽  
pp. 2638-2645 ◽  
Author(s):  
Charlotte Michaux ◽  
Maurizio Sanguinetti ◽  
Fany Reffuveille ◽  
Yanick Auffray ◽  
Brunella Posteraro ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Galleria Mellonella ◽  
Peritoneal Macrophages ◽  
Bacterial Virulence ◽  
Transcriptional Analysis ◽  
Infection Model ◽  
Wild Type ◽  
Content Type ◽  
Wild Type Parent

ABSTRACTPhylogenetic analysis of the crystal structure of theEnterococcus faecalisSlyA (EF_3002) transcriptional factor places it between the SlyA and MarR regulator subfamilies. Proteins of these families are often involved in the regulation of genes important for bacterial virulence and stress response. To gather evidence for the role of this putative regulator inE. faecalisbiology, we dissected the genetic organization of theslyA-EF_3001 locus and constructed aslyAdeletion mutant as well as complemented strains. Interestingly, compared to the wild-type parent, the ΔslyAmutant is more virulent in an insect infection model (Galleria mellonella), exhibits increased persistence in mouse kidneys and liver, and survives better inside peritoneal macrophages. In order to identify a possible SlyA regulon, global microarray transcriptional analysis was performed. This study revealed that theslyA-EF_3001 locus appears to be autoregulated and that 117 genes were differentially regulated in the ΔslyAmutant. In the mutant strain, 111 were underexpressed and 6 overexpressed, indicating that SlyA functions mainly as an activator of transcription.

scholarly journals AbaM Regulates Quorum Sensing, Biofilm Formation and Virulence in Acinetobacter baumannii

2021 ◽  
Author(s):  
Mario López-Martín ◽  
Jean-Frédéric Dubern ◽  
Morgan R. Alexander ◽  
Paul Williams
Keyword(s):  
Quorum Sensing ◽  
Acinetobacter Baumannii ◽  
Biofilm Formation ◽  
Plant Pathogens ◽  
Galleria Mellonella ◽  
Homoserine Lactone ◽  
Infection Model ◽  
Wild Type ◽  
Surface Motility ◽  
Biofilm Development

Acinetobacter baumannii possesses a single divergent luxR/luxI-type quorum sensing (QS) locus named abaR/abaI. This locus also contains a third gene located between abaR and abaI which we term abaM that codes for an uncharacterized member of the RsaM protein family known to regulate N-acylhomoserine lactone (AHL) dependent QS in other β- and γ-proteobacteria. Here we show that disruption of abaM via a T26 insertion in A. baumannii strain AB5075 resulted in increased production of N-(3-hydroxydodecanoyl)-L-homoserine lactone (OHC12) and enhanced surface motility and biofilm formation. In contrast to the wild type and abaI::T26 mutant, the virulence of the abaM::T26 mutant was completely attenuated in a Galleria mellonella infection model. Transcriptomic analysis of the abaM::T26 mutant revealed that AbaM differentially regulates at least 76 genes including the csu pilus operon and the acinetin 505 lipopeptide biosynthetic operon, that are involved in surface adherence, biofilm formation and virulence. A comparison of the wild type, abaM::T26 and abaI::T26 transcriptomes, indicates that AbaM regulates ∼21% of the QS regulon including the csu operon. Moreover, the QS genes (abaI/abaR) were among the most upregulated in the abaM::T26 mutant. A. baumannii lux-based abaM reporter gene fusions revealed that abaM expression is positively regulated by QS but negatively auto-regulated. Overall, the data presented in this work demonstrates that AbaM plays a central role in regulating A. baumannii QS, virulence, surface motility and biofilm formation. Importance Acinetobacter baumanni is a multi-antibiotic resistant pathogen of global healthcare importance. Understanding Acinetobacter virulence gene regulation could aid the development of novel anti-infective strategies. In A. baumannii, the abaR and abaI genes that code for the receptor and synthase components of an N-acylhomoserine (AHL) lactone-dependent quorum sensing system (QS) are separated by abaM. Here we show that although mutation of abaM increased AHL production, surface motility and biofilm development, it resulted in the attenuation of virulence. AbaM was found to control both QS-dependent and QS-independent genes. The significance of this work lies in the identification of AbaM, an RsaM ortholog known to control virulence in plant pathogens, as a modulator of virulence in a human pathogen.

scholarly journals FlhA Influences Bacillus thuringiensis PlcR-Regulated Gene Transcription, Protein Production, and Virulence

2005 ◽  
Vol 71 (12) ◽  
pp. 8903-8910 ◽  
Author(s):  
Laurent Bouillaut ◽  
Nalini Ramarao ◽  
Christophe Buisson ◽  
Nathalie Gilois ◽  
Michel Gohar ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Virulence Factors ◽  
Gene Transcription ◽  
Galleria Mellonella ◽  
Flagellar Apparatus ◽  
Flagellar Proteins ◽  
Crystal Toxins ◽  
Opportunistic Human Pathogen ◽  
Hemolysin Bl ◽  
Lower Production

ABSTRACT Bacillus thuringiensis and Bacillus cereus are closely related. B. thuringiensis is well known for its entomopathogenic properties, principally due to the synthesis of plasmid-encoded crystal toxins. B. cereus appears to be an emerging opportunistic human pathogen. B. thuringiensis and B. cereus produce many putative virulence factors which are positively controlled by the pleiotropic transcriptional regulator PlcR. The inactivation of plcR decreases but does not abolish virulence, indicating that additional factors like flagella may contribute to pathogenicity. Therefore, we further analyzed a mutant (B. thuringiensis 407 Cry− ΔflhA) previously described as being defective in flagellar apparatus assembly and in motility as well as in the production of hemolysin BL and phospholipases. A large picture of secreted proteins was obtained by two-dimensional electrophoresis analysis, which revealed that flagellar proteins are not secreted and that production of several virulence-associated factors is reduced in the flhA mutant. Moreover, we quantified the effect of FlhA on plcA and hblC gene transcription. The results show that the flhA mutation results in a significant reduction of plcA and hblC transcription. These results indicate that the transcription of several PlcR-regulated virulence factors is coordinated with the flagellar apparatus. Consistently, the flhA mutant also shows a strong decrease in cytotoxicity towards HeLa cells and in virulence against Galleria mellonella larvae following oral and intrahemocoelic inoculation. The decrease in virulence may be due to both a lack of flagella and a lower production of secreted factors. Hence, FlhA appears to be an essential virulence factor with a pleiotropic role.

scholarly journals AbaM Regulates Quorum Sensing, Biofilm Formation and Virulence in Acinetobacter baumannii

2020 ◽  
Author(s):  
Mario López-Martín ◽  
Jean-Frédéric Dubern ◽  
Morgan R. Alexander ◽  
Paul Williams
Keyword(s):  
Quorum Sensing ◽  
Acinetobacter Baumannii ◽  
Biofilm Formation ◽  
Plant Pathogens ◽  
Galleria Mellonella ◽  
Homoserine Lactone ◽  
Infection Model ◽  
Wild Type ◽  
Surface Motility ◽  
Biofilm Development

ABSTRACTAcinetobacter baumannii possesses a single divergent luxR/luxI-type quorum sensing (QS) locus named abaR/abaI. This locus also contains a third gene located between abaR and abaI which we term abaM that codes for an uncharacterized member of the RsaM protein family known to regulate N-acylhomoserine lactone (AHL) dependent QS in other β- and γ-proteobacteria. Here we show that disruption of abaM via a T26 insertion in A. baumannii strain AB5075 resulted in increased production of N-(3-hydroxydodecanoyl)-L-homoserine lactone (OHC12) and enhanced surface motility and biofilm formation. In contrast to the wild type and abaI::T26 mutant, the virulence of the abaM::T26 mutant was completely attenuated in a Galleria mellonella infection model. Transcriptomic analysis of the abaM::T26 mutant revealed that abaM differentially regulates at least 76 genes including the csu pilus operon and the acinetin 505 lipopeptide biosynthetic operon, that are involved in surface adherence, biofilm formation and virulence. A comparison of the wild type, abaM::T26 and abaI::T26 transcriptomes, indicates that abaM regulates ~21% of the QS regulon including the csu operon. Moreover, the QS genes (abaI/abaR) were among the most upregulated in the abaM::T26 mutant. A. baumannii lux-based abaM reporter gene fusions revealed that abaM expression is positively regulated by QS but negatively auto-regulated. Overall, the data presented in this work demonstrates that abaM plays a central role in regulating A. baumannii QS, virulence, surface motility and biofilm formation.IMPORTANCEAcinetobacter baumanni is a multi-antibiotic resistant pathogen of global healthcare importance. Understanding Acinetobacter virulence gene regulation could aid the development of novel anti-infective strategies. In A. baumannii, the abaR and abaI genes that code for the receptor and synthase components of an N-acylhomoserine (AHL) lactone-dependent quorum sensing system (QS) are separated by abaM. Here we show that although mutation of abaM increased AHL production, surface motility and biofilm development, it resulted in the attenuation of virulence. abaM was found to control both QS-dependent and QS-independent genes. The significance of this work lies in the identification of abaM, an RsaM ortholog known to control virulence in plant pathogens, as a modulator of virulence in a human pathogen.

scholarly journals Four Superoxide Dismutases Contribute to Bacillus anthracis Virulence and Provide Spores with Redundant Protection from Oxidative Stress

2008 ◽  
Vol 77 (1) ◽  
pp. 274-285 ◽  
Author(s):  
Robert J. Cybulski ◽  
Patrick Sanz ◽  
Farhang Alem ◽  
Scott Stibitz ◽  
Robert L. Bull ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Bacillus Anthracis ◽  
Lethal Dose ◽  
Superoxide Dismutases ◽  
Infection Model ◽  
Wild Type ◽  
Double Knockout ◽  
Sod Activity ◽  
Knockout Strain ◽  
Key Enzyme

ABSTRACT The Bacillus anthracis genome encodes four superoxide dismutases (SODs), enzymes capable of detoxifying oxygen radicals. That two of these SODs, SOD15 and SODA1, are present in the outermost layers of the B. anthracis spore is indicated by previous proteomic analyses of the exosporium. Given the requirement that spores must survive interactions with reactive oxygen species generated by cells such as macrophages during infection, we hypothesized that SOD15 and SODA1 protect the spore from oxidative stress and contribute to the pathogenicity of B. anthracis. To test these theories, we constructed a double-knockout (Δsod15 ΔsodA1) mutant of B. anthracis Sterne strain 34F2 and assessed its lethality in an A/J mouse intranasal infection model. The 50% lethal dose of the Δsod15 ΔsodA1 strain was similar to that of the wild type (34F2), but surprisingly, measurable whole-spore SOD activity was greater than that in 34F2. A quadruple-knockout strain (Δsod15 ΔsodA1 ΔsodC ΔsodA2) was then generated, and as anticipated, spore-associated SOD activity was diminished. Moreover, the quadruple-knockout strain, compared to the wild type, was attenuated more than 40-fold upon intranasal challenge of mice. Spore resistance to exogenously generated oxidative stress and to macrophage-mediated killing correlated with virulence in A/J mice. Allelic exchange that restored sod15 and sodA1 to their wild-type state restored wild-type characteristics. We conclude that SOD molecules within the spore afford B. anthracis protection against oxidative stress and enhance the pathogenicity of B. anthracis in the lung. We also surmise that the presence of four SOD alleles within the genome provides functional redundancy for this key enzyme.

scholarly journals RhlR, but not RhlI, allowsP. aeruginosabacteria to evadeDrosophilaTep4-mediated opsonization

2017 ◽  
Author(s):  
Samantha Haller ◽  
Adrien Franchet ◽  
Abdul Hakkim ◽  
Jing Chen ◽  
Eliana Drenkard ◽  
...  
Keyword(s):  
Intestinal Barrier ◽  
Infection Model ◽  
Dependent Manner ◽  
Intestinal Infection ◽  
Wild Type ◽  
Independent Function ◽  
C Elegans ◽  
Injury Model ◽  
Mutant Phenotypes ◽  
Cellular Immune

ABSTRACTWhenDrosophilaflies feed onPseudomonas aeruginosastrain PA14, some bacteria cross the intestinal barrier and start proliferating inside the hemocoel. This process is limited by hemocytes through phagocytosis. We have previously shown that the PA14 quorum-sensing regulator RhlR is required for these bacteria to elude the cellular immune response. RhlI synthesizes the auto-inducer signal that activates RhlR. Here, we compare the null mutant phenotypes ofrhlRandrhlIin a variety of infection assays inDrosophilaand in the nematodeCaenorhabditis elegans. Surprisingly, inDrosophila, unlikeΔrhlRmutants,ΔrhlImutants are only modestly attenuated for virulence and are poorly phagocytosed and opsonized in a Thioester-containing Protein4-dependent manner. Likewise, ΔrhlIbut not ΔrhlRmutants colonize the digestive tract ofC. elegansand kill it as efficiently as wild-type PA14. Thus, RhlR has an RhlI-independent function in eluding detection or counter-acting the action of the immune system. In contrast to the intestinal infection model,Tep4mutant flies are more resistant to PA14 in a septic injury model, which also depends onrhlR. Thus, the Tep4 putative opsonin can either be protective or detrimental to host defense depending on the infection route.

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