scholarly journals INFLUENCE OF YERSINIA PESTIS PLASMID CONTENT ON BIOFILM FORMATION IN FLEAS WITH DIFFERENT VECTOR ACTIVITY

2018 ◽  
pp. 76-83
Author(s):  
L. P. Bazanova ◽  
E. G. Tokmakova ◽  
G. A. Voronova ◽  
S. V. Balakhonov
Keyword(s):  
Yersinia Pestis ◽  
Biofilm Formation ◽  
Death Rate ◽  
Plasmid Content ◽  
Xenopsylla Cheopis ◽  
Highly Active ◽  
Individual Number ◽  
Vector Activity ◽  
The Individual

Aim. Influence of the plague agent plasmid content on biofilm formation in vivo and death rate of fleas-vectors with different vector activity in experiment were analyzed. Materials and methods. Three Yersinia pestis strains: virulent I-3230 (pYT, pYV, pYP) and I-2638 (pYT, pYV, pYP, pTP 33), and its selected avirulent isogenic clone I-3480 lacking two plasmids (pYV, pYP) were used. Three species of fleas were artificially infected: 477 individuals of Xenopsylla cheopis (a highly active vector), 441 - Citellophilus tesquorum (an active vector), 519 - Frontopsylla lucu-lenta (a low-active vector). The peculiarities of Y. pestis biofilm formation in fleas were estimated by a portion of individuals with bacterial «conglomerates» and «blocks» for a feeding. Death rate of the insects was defined by the percent of the dead fleas at each feeding. Results. All three flea species infected by Y. pestis strains carrying an additional plasmid pTP33 (I-2638 and I-3480) demonstrated the increase of the individual number with various biofilm forms in comparison with the three-plasmid strain I-3230. In X. cheopis it occurred due to the blocked insects, in C. tesquo-rum - mainly due to the fleas containing «conglomerates», in F. luculenta it was completely connected with ectoparasites with «conglomerates». A share of X. cheopis and C. tesquorum died at a feeding was higher in ectoparasites infected with I-3230 strain and F. luculenta - infected by I-2638. Conclusion. Y. pestis strains possessing an additional replicon pTP33 formed a biofilm in the infected insects more often and larger size than a classical three-plasmid variant. Influence of the strain plasmid content on death rate of the infected fleas depended on a vector species.

scholarly journals Identification of gmhA, a Yersinia pestis Gene Required for Flea Blockage, by Using a Caenorhabditis elegans Biofilm System

2005 ◽  
Vol 73 (11) ◽  
pp. 7236-7242 ◽  
Author(s):  
Creg Darby ◽  
Sandya L. Ananth ◽  
Li Tan ◽  
B. Joseph Hinnebusch
Keyword(s):  
Caenorhabditis Elegans ◽  
Yersinia Pestis ◽  
Biofilm Formation ◽  
Yersinia Pseudotuberculosis ◽  
C Elegans ◽  
Transposon Insertion ◽  
Insertion Mutants ◽  
Random Screening

ABSTRACT Yersinia pestis, the cause of bubonic plague, blocks feeding by its vector, the flea. Recent evidence indicates that blockage is mediated by an in vivo biofilm. Y. pestis and the closely related Yersinia pseudotuberculosis also make biofilms on the cuticle of the nematode Caenorhabditis elegans, which block this laboratory animal's feeding. Random screening of Y. pseudotuberculosis transposon insertion mutants with a C. elegans biofilm assay identified gmhA as a gene required for normal biofilms. gmhA encodes phosphoheptose isomerase, an enzyme required for synthesis of heptose, a conserved component of lipopolysaccharide and lipooligosaccharide. A Y. pestis gmhA mutant was constructed and was severely defective for C. elegans biofilm formation and for flea blockage but only moderately defective in an in vitro biofilm assay. These results validate use of the C. elegans biofilm system to identify genes and pathways involved in Y. pestis flea blockage.

scholarly journals Serotype Differences and Lack of Biofilm Formation Characterize Yersinia pseudotuberculosis Infection of the Xenopsylla cheopis Flea Vector of Yersinia pestis

2006 ◽  
Vol 188 (3) ◽  
pp. 1113-1119 ◽  
Author(s):  
David L. Erickson ◽  
Clayton O. Jarrett ◽  
Brendan W. Wren ◽  
B. Joseph Hinnebusch
Keyword(s):  
Yersinia Pestis ◽  
Biofilm Formation ◽  
Yersinia Pseudotuberculosis ◽  
Ex Vivo ◽  
Blood Feeding ◽  
Genetic Change ◽  
Xenopsylla Cheopis ◽  
C Elegans ◽  
Transmissible Infection

ABSTRACT Yersinia pestis, the agent of plague, is usually transmitted by fleas. To produce a transmissible infection, Y. pestis colonizes the flea midgut and forms a biofilm in the proventricular valve, which blocks normal blood feeding. The enteropathogen Yersinia pseudotuberculosis, from which Y. pestis recently evolved, is not transmitted by fleas. However, both Y. pestis and Y. pseudotuberculosis form biofilms that adhere to the external mouthparts and block feeding of Caenorhabditis elegans nematodes, which has been proposed as a model of Y. pestis-flea interactions. We compared the ability of Y. pestis and Y. pseudotuberculosis to infect the rat flea Xenopsylla cheopis and to produce biofilms in the flea and in vitro. Five of 18 Y. pseudotuberculosis strains, encompassing seven serotypes, including all three serotype O3 strains tested, were unable to stably colonize the flea midgut. The other strains persisted in the flea midgut for 4 weeks but did not increase in numbers, and none of the 18 strains colonized the proventriculus or produced a biofilm in the flea. Y. pseudotuberculosis strains also varied greatly in their ability to produce biofilms in vitro, but there was no correlation between biofilm phenotype in vitro or on the surface of C. elegans and the ability to colonize or block fleas. Our results support a model in which a genetic change in the Y. pseudotuberculosis progenitor of Y. pestis extended its pre-existing ex vivo biofilm-forming ability to the flea gut environment, thus enabling proventricular blockage and efficient flea-borne transmission.

scholarly journals Bispecific Antibodies Targeting Different Epitopes on the HIV-1 Envelope Exhibit Broad and Potent Neutralization

2015 ◽  
Vol 89 (24) ◽  
pp. 12501-12512 ◽  
Author(s):  
M. Asokan ◽  
R. S. Rudicell ◽  
M. Louder ◽  
K. McKee ◽  
S. O'Dell ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Bispecific Antibody ◽  
Rhesus Macaques ◽  
Bispecific Antibodies ◽  
Pharmacokinetic Parameters ◽  
Highly Active ◽  
Antibody Specificities ◽  
The Individual ◽  
Hiv 1

ABSTRACTThe potency and breadth of the recently isolated neutralizing human monoclonal antibodies to HIV-1 have stimulated interest in their use to prevent or to treat HIV-1 infection. Due to the antigenically diverse nature of the HIV-1 envelope (Env), no single antibody is highly active against all viral strains. While the physical combination of two broadly neutralizing antibodies (bNAbs) can improve coverage against the majority of viruses, the clinical-grade manufacturing and testing of two independent antibody products are time and resource intensive. In this study, we constructed bispecific immunoglobulins (IgGs) composed of independent antigen-binding fragments with a common Fc region. We developed four different bispecific IgG variants that included antibodies targeting four major sites of HIV-1 neutralization. We show that these bispecific IgGs display features of both antibody specificities and, in some cases, display improved coverage over the individual parental antibodies. All four bispecific IgGs neutralized 94% to 97% of antigenically diverse viruses in a panel of 206 HIV-1 strains. Among the bispecific IgGs tested, VRC07 × PG9-16 displayed the most favorable neutralization profile. It was superior in breadth to either of the individual antibodies, neutralizing 97% of viruses with a median 50% inhibitory concentration (IC50) of 0.055 μg/ml. This bispecific IgG also demonstratedin vivopharmacokinetic parameters comparable to those of the parental bNAbs when administered to rhesus macaques. These results suggest that IgG-based bispecific antibodies are promising candidates for the prevention and treatment of HIV-1 infection in humans.IMPORTANCETo prevent or treat HIV-1 infection, antibodies must potently neutralize nearly all strains of HIV-1. Thus, the physical combination of two or more antibodies may be needed to broaden neutralization coverage and diminish the possibility of viral resistance. A bispecific antibody that has two different antibody binding arms could potentially display neutralization characteristics better than those of any single parental antibody. Here we show that bispecific antibodies contain the binding specificities of the two parental antibodies and that a single bispecific antibody can neutralize 97% of viral strains with a high overall potency. These findings support the use of bispecific antibodies for the prevention or treatment of HIV-1 infection.

scholarly journals BIOFILM FORMATION BY YERSINIA PESTIS STRAINS OF THE MAIN AND NON- MAIN SUBSPECIES FROM NATURAL PLAGUE FOCI OF CENTRAL ASIA IN THE BODY OF XENOPSYLLA CHEOPIS FLEAS

2020 ◽  
pp. 32-38
Author(s):  
L.P. Bazanova ◽  
◽  
E.G. Tokmakova ◽  
G.A. Voronova ◽  
◽  
...  
Keyword(s):  
Central Asia ◽  
Yersinia Pestis ◽  
Biofilm Formation ◽  
The Body ◽  
Aggregation Index ◽  
Xenopsylla Cheopis ◽  
Blood Sucking ◽  
Central Asian ◽  
Natural Foci

The ability to form biofilms (“bacterial lumps” and “blocks”) by strains of different subspecies of Y. pestis in the body of the flea X. cheopis was studied. It was found that strains of the main subspecies form a biofilm almost three times more often than strains of the non-main subspecies – altaica and ulegeica (14.8±2.50 vs. 5.4±1.10). These differences were mainly due to the formation of “blocks” (9.2±1.66 vs. 1.6±0.45), rather than “bacterial lumps” (5.6±1.40 vs. 3.8±0.83). For X. cheopis fleas, infection with strains of the main subspecies in most cases leads to the predominance of massive forms of biofilm – “blocks” – aggregation index >1. When infecting with non-main subspecies pathogen from Central Asian natural foci, the persistence of the microbe in X. cheopis fleas usually occurs in the form of “lumps” and the aggregation index does not exceed 1. The success of transmission by fleas of this species is ensured not by the total number of blocked individuals, but by their presence and, accordingly, by probable blood-sucking during feeding.

Transmission of Yersinia pestis cultures with different plasmid content from Xenopsylla cheopis to Calomys callosus

2003 ◽  
Vol 89 (3) ◽  
pp. 159-162 ◽  
Author(s):  
A. de Almeida ◽  
L. Alves ◽  
R. Amaral ◽  
W. França ◽  
N. Leal
Keyword(s):  
Yersinia Pestis ◽  
Plasmid Content ◽  
Xenopsylla Cheopis ◽  
Calomys Callosus

scholarly journals In VitroAnalyses of the Effects of Heparin and Parabens on Candida albicans Biofilms and Planktonic Cells

2011 ◽  
Vol 56 (1) ◽  
pp. 148-153 ◽  
Author(s):  
Marisa H. Miceli ◽  
Stella M. Bernardo ◽  
T. S. Neil Ku ◽  
Carla Walraven ◽  
Samuel A. Lee
Keyword(s):  
Candida Albicans ◽  
Metabolic Activity ◽  
Biofilm Formation ◽  
High Dose ◽  
Dependent Manner ◽  
Planktonic Cells ◽  
Content Type ◽  
Heparin Sodium ◽  
The Individual

ABSTRACTInfections and thromboses are the most common complications associated with central venous catheters. Suggested strategies for prevention and management of these complications include the use of heparin-coated catheters, heparin locks, and antimicrobial lock therapy. However, the effects of heparin onCandida albicansbiofilms and planktonic cells have not been previously studied. Therefore, we sought to determine thein vitroeffect of a heparin sodium preparation (HP) on biofilms and planktonic cells ofC. albicans. Because HP contains two preservatives, methyl paraben (MP) and propyl paraben (PP), these compounds and heparin sodium without preservatives (Pure-H) were also tested individually. The metabolic activity of the mature biofilm after treatment was assessed using XTT [2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] reduction and microscopy. Pure-H, MP, and PP caused up to 75, 85, and 60% reductions of metabolic activity of the mature preformedC. albicansbiofilms, respectively. Maximal efficacy against the mature biofilm was observed with HP (up to 90%) compared to the individual compounds (P< 0.0001). Pure-H, MP, and PP each inhibitedC. albicansbiofilm formation up to 90%. A complete inhibition of biofilm formation was observed with HP at 5,000 U/ml and higher. When tested against planktonic cells, each compound inhibited growth in a dose-dependent manner. These data indicated that HP, MP, PP, and Pure-H havein vitroantifungal activity againstC. albicansmature biofilms, formation of biofilms, and planktonic cells. Investigation of high-dose heparin-based strategies (e.g., heparin locks) in combination with traditional antifungal agents for the treatment and/or prevention ofC. albicansbiofilms is warranted.

scholarly journals Preclinical Testing of Radiopharmaceuticals for the Detection and Characterization of Osteomyelitis: Experiences from a Porcine Model

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4221
Author(s):  
Aage Kristian Olsen Alstrup ◽  
Svend Borup Jensen ◽  
Ole Lerberg Nielsen ◽  
Lars Jødal ◽  
Pia Afzelius
Keyword(s):  
Animal Model ◽  
Animal Models ◽  
Porcine Model ◽  
Animal Experiments ◽  
Radioactive Tracers ◽  
The Individual ◽  
Selection Of

The development of new and better radioactive tracers capable of detecting and characterizing osteomyelitis is an ongoing process, mainly because available tracers lack selectivity towards osteomyelitis. An integrated part of developing new tracers is the performance of in vivo tests using appropriate animal models. The available animal models for osteomyelitis are also far from ideal. Therefore, developing improved animal osteomyelitis models is as important as developing new radioactive tracers. We recently published a review on radioactive tracers. In this review, we only present and discuss osteomyelitis models. Three ethical aspects (3R) are essential when exposing experimental animals to infections. Thus, we should perform experiments in vitro rather than in vivo (Replacement), use as few animals as possible (Reduction), and impose as little pain on the animal as possible (Refinement). The gain for humans should by far exceed the disadvantages for the individual experimental animal. To this end, the translational value of animal experiments is crucial. We therefore need a robust and well-characterized animal model to evaluate new osteomyelitis tracers to be sure that unpredicted variation in the animal model does not lead to a misinterpretation of the tracer behavior. In this review, we focus on how the development of radioactive tracers relies heavily on the selection of a reliable animal model, and we base the discussions on our own experience with a porcine model.

scholarly journals Biofilm formation in the lung contributes to virulence and drug tolerance of Mycobacterium tuberculosis

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Poushali Chakraborty ◽  
Sapna Bajeli ◽  
Deepak Kaushal ◽  
Bishan Dass Radotra ◽  
Ashwani Kumar
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Immune System ◽  
Biofilm Formation ◽  
Antimycobacterial Activity ◽  
Drug Tolerance ◽  
Host Immune System ◽  
Tissue Sections ◽  
Slow Growing

AbstractTuberculosis is a chronic disease that displays several features commonly associated with biofilm-associated infections: immune system evasion, antibiotic treatment failures, and recurrence of infection. However, although Mycobacterium tuberculosis (Mtb) can form cellulose-containing biofilms in vitro, it remains unclear whether biofilms are formed during infection in vivo. Here, we demonstrate the formation of Mtb biofilms in animal models of infection and in patients, and that biofilm formation can contribute to drug tolerance. First, we show that cellulose is also a structural component of the extracellular matrix of in vitro biofilms of fast and slow-growing nontuberculous mycobacteria. Then, we use cellulose as a biomarker to detect Mtb biofilms in the lungs of experimentally infected mice and non-human primates, as well as in lung tissue sections obtained from patients with tuberculosis. Mtb strains defective in biofilm formation are attenuated for survival in mice, suggesting that biofilms protect bacilli from the host immune system. Furthermore, the administration of nebulized cellulase enhances the antimycobacterial activity of isoniazid and rifampicin in infected mice, supporting a role for biofilms in phenotypic drug tolerance. Our findings thus indicate that Mtb biofilms are relevant to human tuberculosis.

scholarly journals Eap1p, an Adhesin That Mediates Candida albicans Biofilm Formation In Vitro and In Vivo

2007 ◽  
Vol 6 (6) ◽  
pp. 931-939 ◽  
Author(s):  
Fang Li ◽  
Michael J. Svarovsky ◽  
Amy J. Karlsson ◽  
Joel P. Wagner ◽  
Karen Marchillo ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Cell Adhesion ◽  
Biofilm Formation ◽  
Fungal Infections ◽  
Gene Dosage ◽  
Venous Catheter ◽  
Flow Chamber ◽  
Dependent Manner

ABSTRACT Candida albicans is the leading cause of systemic fungal infections in immunocompromised humans. The ability to form biofilms on surfaces in the host or on implanted medical devices enhances C. albicans virulence, leading to antimicrobial resistance and providing a reservoir for infection. Biofilm formation is a complex multicellular process consisting of cell adhesion, cell growth, morphogenic switching between yeast form and filamentous states, and quorum sensing. Here we describe the role of the C. albicans EAP1 gene, which encodes a glycosylphosphatidylinositol-anchored, glucan-cross-linked cell wall protein, in adhesion and biofilm formation in vitro and in vivo. Deleting EAP1 reduced cell adhesion to polystyrene and epithelial cells in a gene dosage-dependent manner. Furthermore, EAP1 expression was required for C. albicans biofilm formation in an in vitro parallel plate flow chamber model and in an in vivo rat central venous catheter model. EAP1 expression was upregulated in biofilm-associated cells in vitro and in vivo. Our results illustrate an association between Eap1p-mediated adhesion and biofilm formation in vitro and in vivo.

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