scholarly journals Heterologous Expression of the Pathogen-Specific LIC11711 Gene in the Saprophyte L. biflexa Increases Bacterial Binding to Laminin and Plasminogen

Pathogens ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 599
Author(s):  
Leandro Toshio Kochi ◽  
Luis Guilherme Virgílio Fernandes ◽  
Ana Lucia Tabet Oller Nascimento
Keyword(s):  
Molecular Biology ◽  
Heterologous Expression ◽  
Pathogenic Bacteria ◽  
Bacterial Pathogenesis ◽  
Bacterial Virulence ◽  
Febrile Disease ◽  
Bacterial Binding ◽  
Etiological Agents ◽  
First Time

Leptospirosis is a febrile disease and the etiological agents are pathogenic bacteria of the genus Leptospira. The leptospiral virulence mechanisms are not fully understood and the application of genetic tools is still limited, despite advances in molecular biology techniques. The leptospiral recombinant protein LIC11711 has shown interaction with several host components, indicating a potential function in virulence. This study describes a system for heterologous expression of the L. interrogans gene lic11711 using the saprophyte L. biflexa serovar Patoc as a surrogate, aiming to investigate its possible activity in bacterial virulence. Heterologous expression of LIC11711 was performed using the pMaOri vector under regulation of the lipL32 promoter. The protein was found mainly on the leptospiral outer surface, confirming its location. The lipL32 promoter enhanced the expression of LIC11711 in L. biflexa compared to the pathogenic strain, indicating that this strategy may be used to overexpress low-copy proteins. The presence of LIC11711 enhanced the capacity of L. biflexa to adhere to laminin (Lam) and plasminogen (Plg)/plasmin (Pla) in vitro, suggesting the involvement of this protein in bacterial pathogenesis. We show for the first time that the expression of LIC11711 protein of L. interrogans confers a virulence-associated phenotype on L. biflexa, pointing out possible mechanisms used by pathogenic leptospires.

scholarly journals Sensitivity of Vermamoeba (Hartmannella) vermiformis cysts to conventional disinfectants and protease

2014 ◽  
Vol 13 (2) ◽  
pp. 302-310 ◽  
Author(s):  
Emilie Fouque ◽  
Yann Héchard ◽  
Philippe Hartemann ◽  
Philippe Humeau ◽  
Marie-Cécile Trouilhé
Keyword(s):  
Legionella Pneumophila ◽  
Pathogenic Bacteria ◽  
Hot Water ◽  
Water Networks ◽  
Important Health ◽  
Hartmannella Vermiformis ◽  
Important Health Issue ◽  
Few Data ◽  
First Time

Vermamoeba vermiformis is a free-living amoeba (FLA) widely distributed in the environment, known to colonize hot water networks and to be the reservoir of pathogenic bacteria such as Legionella pneumophila. FLA are partly resistant to biocides, especially in their cyst form. The control of V. vermiformis in hot water networks represents an important health issue, but there are very few data on their resistance to disinfection treatments. The sensitivity of cysts of two strains of V. vermiformis to three disinfectants frequently used in hot water networks (chlorine, heat shock, peracetic acid (PAA) mixed with hydrogen peroxide (H2O2)) was investigated. In vitro, several concentrations of biocides, temperatures and exposure times according to the French regulation were tested. Cysts were fully inactivated by the following conditions: 15 mg/L of chlorine for 10 min; 60 °C for 30 min; and 0.5 g/L equivalent H2O2 of PAA mixed with H2O2 for 30 min. For the first time, the strong efficacy of subtilisin (0.625 U/mL for 24 h), a protease, to inactivate the V. vermiformis cysts has been demonstrated. It suggests that novel approaches may be efficient for disinfection processes. Finally, V. vermifomis cysts were sensitive to all the tested treatments and appeared to be more sensitive than Acanthamoeba cysts.

scholarly journals Plasminogen Acquisition and Activation at the Surface of Leptospira Species Lead to Fibronectin Degradation

2009 ◽  
Vol 77 (9) ◽  
pp. 4092-4101 ◽  
Author(s):  
Monica L. Vieira ◽  
Silvio A. Vasconcellos ◽  
Amane P. Gonçales ◽  
Zenaide M. de Morais ◽  
Ana L. T. O. Nascimento
Keyword(s):  
Aminocaproic Acid ◽  
Cellular Growth ◽  
Activation System ◽  
Lysine Residues ◽  
Etiological Agents ◽  
Low Passage ◽  
First Time ◽  
Dose Dependent ◽  
Leptospira Species

ABSTRACT Pathogenic Leptospira species are the etiological agents of leptospirosis, a widespread disease of human and veterinary concern. In this study, we report that Leptospira species are capable of binding plasminogen (PLG) in vitro. The binding to the leptospiral surface was demonstrated by indirect immunofluorescence confocal microscopy with living bacteria. The PLG binding to the bacteria seems to occur via lysine residues because the ligation is inhibited by addition of the lysine analog 6-aminocaproic acid. Exogenously provided urokinase-type PLG activator (uPA) converts surface-bound PLG into enzymatically active plasmin, as evaluated by the reaction with the chromogenic plasmin substrate d-Val-Leu-Lys 4-nitroanilide dihydrochloridein. The PLG activation system on the surface of Leptospira is PLG dose dependent and does not cause injury to the organism, as cellular growth in culture was not impaired. The generation of active plasmin within Leptospira was observed with several nonvirulent high-passage strains and with the nonpathogenic saprophytic organism Leptospira biflexa. Statistically significant higher activation of plasmin was detected with a low-passage infectious strain of Leptospira. Plasmin-coated virulent Leptospira interrogans bacteria were capable of degrading purified extracellular matrix fibronectin. The breakdown of fibronectin was not observed with untreated bacteria. Our data provide for the first time in vitro evidence for the generation of active plasmin on the surface of Leptospira, a step that may contribute to leptospiral invasiveness.

scholarly journals Minimalistic mycoplasmas harbor different functional toxin-antitoxin systems

2021 ◽  
Author(s):  
Virginia Hill ◽  
Hatice Akarsu ◽  
Rubén Sánchez Barbarroja ◽  
Valentina Cippà ◽  
Martin Heller ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
In Silico ◽  
Model Organism ◽  
Future Research ◽  
Phylogenetic Distance ◽  
Minimal Cell ◽  
Unequal Distribution ◽  
Mycoplasma Mycoides ◽  
First Time

AbstractMycoplasmas are minute bacteria controlled by very small genomes ranging from 0.6 to 1.4 Mbp. They lack a cell wall and have been suggested to have progressed through reductive evolution from phylogenetically closely related Clostridia. They are known to colonize the respiratory tract or the urogenital tract among other organs and can cause chronic and subclinical diseases associated with long persistence of the causative agent. Toxin-antitoxin systems (TAS) are genetic elements that have been described for several respiratory and urogenital pathogens as well as for Clostridia, but never for pathogenic mycoplasmas. Here we describe for the first-time different types of TAS in a Mycoplasma pathogen, namely M. mycoides subsp. capri. We identified candidate TAS in silico via TASmania database. Two candidate TAS identified in silico and another candidate TAS suggested in a minimal cell based on transposon mutagenesis were systematically tested for their functionality in hosts with different phylogenetic distance using heterologous expression. Phylogenetic distance of the host used for heterologous expression influenced the outcome of the functional testing. We corroborated functionality of the three candidate TAS in Mycoplasma capricolum subsp. capricolum. Moreover, we confirmed transcription and translation of molecules of the TAS investigated during in vitro growth. We sequence analyzed 15 genomes of M. mycoides subsp. capri and revealed an unequal distribution of the TAS studied pointing towards dynamic gain and loss of TAS within the species.Author summaryMycoplasmas have a minimal genome and have never been shown to possess TAS. In this work we showed the presence of different functional TAS systems in Mycoplasma mycoides subsp. capri, a caprine pathogen for the first time. Sequence analysis of a number of Mycoplasma mycoides subsp. capri strains revealed a plasticity of the genome with respect to TAS carriage. This work paves the way to investigate the biological role of TAS (e.g. persistence, stress tolerance) during infection using mycoplasmas as a simple model organism. Since most mycoplasmas lack classical virulence factors such as exotoxins and go into a kind of stealth mode to evade the immune system, TAS are likely to contribute to the parasitic lifestyle of mycoplasmas and should be investigated in that respect. The availability of synthetic genomics tools to modify a range of Mycoplasma pathogens and well-established challenge models for the latter mycoplasmas will foster future research on TAS in mycoplasmas.

scholarly journals Schwann Cell Autophagy and Necrosis as Mechanisms of Cell Death by Acanthamoeba

Pathogens ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 458
Author(s):  
Ismael Castelan-Ramírez ◽  
Lizbeth Salazar-Villatoro ◽  
Bibiana Chávez-Munguía ◽  
Citlaltepetl Salinas-Lara ◽  
Carlos Sánchez-Garibay ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Confocal Microscopy ◽  
Rattus Norvegicus ◽  
Multilamellar Bodies ◽  
Transmission Electron ◽  
Granulomatous Amoebic Encephalitis ◽  
Etiological Agents ◽  
First Time

Amoebae of the genus Acanthamoeba are etiological agents of granulomatous amoebic encephalitis (GAE). Recently, through an in vivo GAE model, Acanthamoeba trophozoites were immunolocalized in contact with the peripheral nervous system (PNS) cells—Schwann cells (SC). In this study, we analyzed in greater detail the in vitro early morphological events (1, 2, 3, and 4 h) during the interaction of A. culbertsoni trophozoites (ATCC 30171) with SC from Rattus norvegicus (ATCC CRL-2941). Samples were processed for scanning and transmission electron microscopy as well as confocal microscopy. After 1 h of interaction, amoebae were observed to be adhered to the SC cultures, emitting sucker-like structures associated with micro-phagocytic channels. In addition, evidence of necrosis was identified since edematous organelles as well as multivesicular and multilamellar bodies characteristics of autophagy were detected. At 2 h, trophozoites migrated beneath the SC culture in which necrosis and autophagy persisted. By 3 and 4 h, extensive lytic zones were observed. SC necrosis was confirmed by confocal microscopy. We reported for the first time the induction of autophagic and necrotic processes in PNS cells, associated in part with the contact-dependent pathogenic mechanisms of A. culbertsoni trophozoites.

scholarly journals Pathogenicity of bacteria isolated from 'Hua Moa' plantain with bunch abortion symptoms inoculated into other clones of plantain and banana.

1969 ◽  
Vol 91 (1-2) ◽  
pp. 19-30
Author(s):  
Judith Rengifo ◽  
Mildred Zapata ◽  
Manuel Díaz ◽  
Rafael Inglés
Keyword(s):  
Puerto Rico ◽  
Pathogenic Bacteria ◽  
Soft Rot ◽  
Field Studies ◽  
Bacterial Diseases ◽  
Burkholderia Gladioli ◽  
Agar Media ◽  
The Caribbean ◽  
First Time

Plantain production (Musa spp.) in Central and South America and the Caribbean is affected by bacterial diseases. In Puerto Rico, production of the plantain 'Hua Moa' is affected by a condition known as bunch abortion or 'choke neck'. This condition in some cases is accompanied by soft rot symptoms and therefore could be related to phytopathogenic bacteria. The purpose of this study was to identify the bacteria related to the soft rot tissues on plants affected with the bunch abortion symptoms and determine their virulence in other clones of plantain and banana. Bacterial colonies were isolated from pseudostems of Hua Moa at three locations in Puerto Rico, using semi-selective and nutritive agar media. The pathogenicity was determined by using potato and plantain pseudostem discs in humid chambers under in vitro conditions. Pathogenic bacteria were identified by using the BIOLOG® system. Under greenhouse conditions, the virulence of Burkholderia gladioli, Pseudomonas spinosa, Erwinia chrysanthemi and Pseudomonas aeruginosa was confirmed on plantain (Maricongo, FIAH-121, Enano Común, and Hua Moa) and banana (Grand Nain) using a scale from 1 to 9. All clones evaluated under greenhouse conditions were susceptible to E. chrysanthemi, potential causal agent of soft rot in Hua Moa plantain once the bunch abortion appears. Burkholderia gladioli is reported for the first time affecting plantain and banana.This bacterium was more virulent in Hua Moa and Grand Nain clones with severity of 7 and 5, respectively, than in Maricongo, severity 3. Pseudomonas spinosa and P. aeruginosa produced less damage with severity less than 4, than E. chrysanthemi and B. gladioli with severity greater than 4. This research was conducted under in vitro and greenhouse conditions and demonstrates that the most important bacteria causing soft rot in plants with the choke neck condition are E. chrysanthemi and B. gladioli. It is recommended to conduct field studies using plants up to the reproductive stage to determine whether these bacteria are also related as causal agents of the choke neck.

Somatic Embryogenesis and In vitro Plant Regeneration in Vigna trilobata (L.) Verd. - A Potential Pasture Legume

1970 ◽  
Vol 19 (1) ◽  
pp. 89-99
Author(s):  
K. Choudhary ◽  
M. Singh ◽  
M. S. Rathore ◽  
N. S. Shekhawat
Keyword(s):  
Somatic Embryogenesis ◽  
Growth Regulators ◽  
Somatic Embryos ◽  
Basal Medium ◽  
Long Term Study ◽  
Continuous Application ◽  
First Time ◽  
Pasture Legume

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l  of 2,4-D and 0.5 mg/l  of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture  legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)

Preliminary in Vitro Investigations on The Inhibitory Activity of The Original Dietary Supplement Oxidal® on Pathogenic Bacterial Strains

2020 ◽  
Vol 11 ◽  
pp. 37-43
Author(s):  
Prof. Teodora P. Popova ◽  
Toshka Petrova ◽  
Ignat Ignatov ◽  
Stoil Karadzhov
Keyword(s):  
Dietary Supplement ◽  
Broad Spectrum ◽  
Pathogenic Bacteria ◽  
Antimicrobial Agents ◽  
Clinical Effectiveness ◽  
Inhibitory Effect ◽  
Diffusion Method ◽  
Bacterial Strains ◽  
Microbial Strains

The antimicrobial action of the dietary supplement Oxidal® was tested using the classic Bauer and Kirby agar-gel diffusion method. Clinical and reference strains of Staphylococcus aureus and Escherichia coli were used in the studies. The tested dietary supplement showed a well-pronounced inhibitory effect against the microbial strains commensurable with that of the broad-spectrum chemotherapeutic agent Enrofloxacin and showed even higher activity than the broad spectrum antibiotic Thiamphenicol. The proven inhibitory effect of the tested dietary supplement against the examined pathogenic bacteria is in accordance with the established clinical effectiveness standards for antimicrobial agents.

THE ACTIVITY OF ESSENTIAL OILS OBTAINED FROM SPECIES AND INTERSPECIES HYBRIDS OF THE Mentha GENUS AGAINST SELECTED PLANT PATHOGENIC BACTERIA

2018 ◽  
Vol 17 (6) ◽  
pp. 167-174 ◽  
Author(s):  
Małgorzata Schollenberger ◽  
Tomasz M. Staniek ◽  
Elżbieta Paduch-Cichal ◽  
Beata Dasiewicz ◽  
Agnieszka Gadomska-Gajadhur ◽  
...  
Keyword(s):  
Essential Oils ◽  
Pseudomonas Syringae ◽  
Pathogenic Bacteria ◽  
Antimicrobial Effect ◽  
Mentha Piperita ◽  
Mentha Spicata ◽  
Plant Essential Oils ◽  
Plant Pathogenic Bacteria ◽  
Interspecies Hybrids

Plant essential oils of six aromatic herb species and interspecies hybrids of the family Lamiaceae – chocolate mint (Mentha piperita × ‘Chocolate’), pineapple mint (Mentha suaveolens ‘Variegata’), apple mint (Mentha × rotundifolia), spearmint (Mentha spicata), orange mint (Mentha × piperita ‘Granada’) and strawberry mint (Mentha × villosa ‘Strawberry’) – were investigated for antimicrobial effects against plant pathogenic bacteria: Agrobacterium tumefaciens, Pseudomonas syringae pv. syringae and Xanthomonas arboricola pv. corylina. The screening was carried out in vitro on agar plates filled with the target organism. All essential oils screened exhibited a higher level of antibacterial activity against A. tumefaciens and X. arboricola pv. corylina than streptomycin used as a standard in all tests. The antimicrobial effect of streptomycin and five mint oils was at the same level for P. syringae pv. syringae. There were no significant differences in the influence of the chocolate mint oil on the growth inhibition of all bacteria tested. Plant essential oils from pineapple mint, apple mint, spearmint and strawberry mint showed the weakest antimicrobial activity against P. syringae pv. syringae and the strongest towards A. tumefaciens and X. arboricola pv. corylina. The essential oils from strawberry mint, pineapple mint, spearmint and apple mint had the strongest effect on A. tumefaciens, and the lowest inhibitory activity was exhibited by the chocolate mint and orange mint essential oils. X. arboricola pv. corylina was the most sensitive to the strawberry mint, pineapple mint and spearmint oils. The chocolate mint oil showed the greatest activity against P. syringae pv. syringae.

Dwindling status of Epimedium Elatum (Morren & Decne) and its geographical distribution in Kashmir Himalaya, India

2018 ◽  
pp. 47-52
Keyword(s):  
Medicinal Plant ◽  
Biological Activities ◽  
Conservation Status ◽  
Natural Populations ◽  
Culture Techniques ◽  
Kashmir Himalayas ◽  
Near Future ◽  
First Time ◽  
Tissue Culture Techniques

Epimedium elatum (Morren & Decne) of family Berberidaceace is a rare perennial medicinal plant, endemic to high altitude forests of Northwestern Himalayas in India. Ethnobotanically, it has been used as an ingredient for treatment of bone-joint disorders, impotence and kidney disorders in Kashmir Himalayas. Phytochemically, it is rich in Epimedin ABC and Icariin; all of these have been demonstrated to possess remarkable biological activities like PDE-5 inhibition (treatment of erectile dysfunction), anticancer, antiosteoporosis antioxidant and antiviral properties. The present investigation reports its traditional usage, comprehensive distribution and conservation status from twenty ecogeographical regions in Kashmir Himalayas, India. The species was reported from Gurez valley for the first time. Numerous threats like excessive grazing, deforestration, habitat fragmentation, tourism encroachment, landslides and excessive exploitation have decreased its natural populations in most of the surveyed habitats. Consequently, its existence may become threatened in near future if timely conservation steps are not taken immediately by concerned stakeholders involved in medicinal plant research. Moreover, use of plant tissue culture techniques is recommended for development of its in vitro propagation protocols. Therefore, introduction of this medicinal plant in botanical gardens, protected sites and development of monitoring programmes are needed for its immediate conservation in Northwestern Himalayas, India.

Structure-Reactivity Relationships on Substrates and Inhibitors of the Lysine Deacylase Sirtuin 2 from Schistosoma Mansoni (SmSirt2)

2019 ◽  
Author(s):  
Daria Monaldi ◽  
Dante Rotili ◽  
Julien Lancelot ◽  
Martin Marek ◽  
Nathalie Wössner ◽  
...  
Keyword(s):  
Schistosoma Mansoni ◽  
Human Cancer ◽  
Egg Laying ◽  
Human Cancer Cells ◽  
Parasite Survival ◽  
Survival And Reproduction ◽  
Emergence Of Resistance ◽  
First Time ◽  
Parasitic Life Cycle

The only drug for treatment of Schistosomiasis is Praziquantel, and the possible emergence of resistance makes research on novel therapeutic agents necessary. Targeting of Schistosoma mansoni epigenetic enzymes, which regulate the parasitic life cycle, emerged as promising approach. Due to the strong effects of human Sirtuin inhibitors on parasite survival and reproduction, Schistosoma sirtuins were postulated as therapeutic targets. In vitro testing of synthetic substrates of S. mansoni Sirtuin 2 (SmSirt2) and kinetic experiments on a myristoylated peptide demonstrated lysine long chain deacylation as an intrinsic SmSirt2 activity for the first time. Focused in vitro screening of the GSK Kinetobox library and structure-activity relationships (SAR) of identified hits, led to the first SmSirt2 inhibitors with activity in the low micromolar range. Several SmSirt2 inhibitors showed potency against both larval schistosomes (viability) and adult worms (pairing, egg laying) in culture without general toxicity to human cancer cells.<br>

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