scholarly journals In Vitro Cultivation of the Syphilis Spirochete Treponema pallidum

2021 ◽  
Vol 1 (2) ◽  
Author(s):  
Diane G. Edmondson ◽  
Steven J. Norris
Keyword(s):  
Treponema Pallidum ◽  
In Vitro Cultivation

scholarly journals Genetic Engineering of Treponema pallidum subsp. pallidum, the Syphilis Spirochete

2021 ◽  
Author(s):  
Emily Romeis ◽  
Lauren Tantalo ◽  
Nicole Lieberman ◽  
Quynh Phung ◽  
Alex Greninger ◽  
...  
Keyword(s):  
Genetic Engineering ◽  
Vaccine Development ◽  
Genetic Manipulation ◽  
Treponema Pallidum ◽  
Gene Promoter ◽  
Digital Pcr ◽  
Kanamycin Resistance ◽  
In Vitro Cultivation

Background.  Despite more than a century of research, genetic manipulation of Treponema pallidum subsp. pallidum ( T. pallidum ), the causative of agent of syphilis, has not been successful. The lack of genetic engineering tools has severely limited understanding the mechanisms behind T. pallidum success as a pathogen. A recently described method for in vitro cultivation of T. pallidum, however, has made possible to experiment with transformation and selection protocols in this pathogen. Here, we describe an approach that successfully replaced the tprA ( tp0009 ) pseudogene in the SS14 T. pallidum strain with a kanamycin resistance ( kan R ) cassette.               Principal findings. A suicide vector was constructed using the pUC57 plasmid backbone. In the vector, the kan R gene was cloned downstream of the tp0574 gene promoter. The tp0574 prom- kan R cassette was then placed between two 1-kbp homology arms identical to the sequences upstream and downstream of the tprA pseudogene. To induce homologous recombination and integration of the kan R cassette into T. pallidum chromosome, in vitro -cultured SS14 strain spirochetes were exposed to the engineered vector in a CaCl 2 -based transformation buffer and let recover for 24 hours before adding kanamycin-containing selective media. Integration of the kan R cassette was demonstrated by qualitative PCR, droplet digital PCR (ddPCR), and whole genome sequencing (WGS) of transformed treponemes propagated in vitro and in vivo . ddPCR analysis of RNA and mass spectrometry confirmed expression of the kan R message and protein in treponemes propagated in vitro . Moreover, tprA knockout ( tprA ko -SS14) treponemes grew in kanamycin concentrations that were 64 times higher than the MIC for the wild-type SS14 (wt-SS14) strain and in infected rabbits treated with kanamycin.             Conclusion. We demonstrated that genetic manipulation of T. pallidum is attainable. This discovery will allow the application of functional genetics techniques to study syphilis pathogenesis and improve syphilis vaccine development.

scholarly journals Analysis of Treponema pallidum Strains From China Using Improved Methods for Whole-Genome Sequencing From Primary Syphilis Chancres

2020 ◽  
Author(s):  
Wentao Chen ◽  
David Šmajs ◽  
Yongfei Hu ◽  
Wujian Ke ◽  
Petra Pospíšilová ◽  
...  
Keyword(s):  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Whole Genome Amplification ◽  
Treponema Pallidum ◽  
Storage Conditions ◽  
In Vitro Cultivation ◽  
Whole Genome ◽  
Primary Syphilis ◽  
Improved Methods

Abstract Background Whole-genome sequencing (WGS) of Treponema pallidum subspecies pallidum (TPA) has been constrained by the lack of in vitro cultivation methods for isolating spirochetes from patient samples. Methods We built upon recently developed enrichment methods to sequence TPA directly from primary syphilis chancre swabs collected in Guangzhou, China. Results By combining parallel, pooled whole-genome amplification with hybrid selection, we generated high-quality genomes from 4 of 8 chancre-swab samples and 2 of 2 rabbit-passaged isolates, all subjected to challenging storage conditions. Conclusions This approach enabled the first WGS of Chinese samples without rabbit passage and provided insights into TPA genetic diversity in China.

scholarly journals The in vitro cultivation of Treponema pallidum. Corroborative studies.

1977 ◽  
Vol 53 (6) ◽  
pp. 338-339
Author(s):  
J W Foster ◽  
D S Kellogg ◽  
J W Clark ◽  
A Balows
Keyword(s):  
Treponema Pallidum ◽  
In Vitro Cultivation

scholarly journals P074 In vitro and in vivo efficacy of linezolid on Treponema pallidum, the syphilis agent

2021 ◽  
Author(s):  
L Giacani ◽  
A Haynes ◽  
M Vall Mayans ◽  
M Ubals Cazorla ◽  
C Nieto ◽  
...  
Keyword(s):  
Treponema Pallidum ◽  
In Vivo Efficacy

scholarly journals A prospectus of plant growth promoting endophytic bacterium from orchid (Vanda cristata)

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sujit Shah ◽  
Krishna Chand ◽  
Bhagwan Rekadwad ◽  
Yogesh S. Shouche ◽  
Jyotsna Sharma ◽  
...  
Keyword(s):  
Plant Growth ◽  
Root Colonization ◽  
Endophytic Bacterium ◽  
Epifluorescence Microscopy ◽  
In Vitro Cultivation ◽  
Rrna Gene ◽  
Bacillus Spp ◽  
Plant Growth Promoting ◽  
Growth Promoting

Abstract Background A plant growth-promoting endophytic bacterium PVL1 isolated from the leaf of Vanda cristata has the ability to colonize with roots of plants and protect the plant. PVL1 was isolated using laboratory synthetic media. 16S rRNA gene sequencing method has been employed for identification before and after root colonization ability. Results Original isolated and remunerated strain from colonized roots were identified as Bacillus spp. as per EzBiocloud database. The presence of bacteria in the root section of the plantlet was confirmed through Epifluorescence microscopy of colonized roots. The in-vitro plantlet colonized by PVL1 as well as DLMB attained higher growth than the control. PVL1 capable of producing plant beneficial phytohormone under in vitro cultivation. HPLC and GC-MS analysis suggest that colonized plants contain Indole Acetic Acid (IAA). The methanol extract of Bacillus spp., contains 0.015 μg in 1 μl concentration of IAA. PVL1 has the ability to produce antimicrobial compounds such as ethyl iso-allocholate, which exhibits immune restoring property. One-way ANOVA shows that results were statistically significant at P ≤ 0.05 level. Conclusions Hence, it has been concluded that Bacillus spp. PVL1 can promote plant growth through secretion of IAA during root colonization and ethyl iso-allocholate to protect plants from foreign infections. Thus, this study supports to support Koch’s postulates of bacteria establishment.

scholarly journals What quantitative mechanical loading stimulates in vitro cultivation best?

2015 ◽  
Vol 2 (1) ◽  
Author(s):  
Jerry Natenstedt ◽  
Aimee C Kok ◽  
Jenny Dankelman ◽  
Gabrielle JM Tuijthof
Keyword(s):  
Mechanical Loading ◽  
In Vitro Cultivation
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