Complete Genome Sequence of Salmonella Phage vB_SenA_SM5, Active against Multidrug-Resistant Salmonella enterica Serovar Typhi Isolates

Chaudhary, Naveen; Singh, Dharminder; Maurya, Ravi Kumar; Mohan, Balvinder; Taneja, Neelam

doi:10.1128/mra.00309-22

Cited by 3 publications

(3 citation statements)

References 9 publications

(9 reference statements)

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“…Of interest, many of these impacted Salmonella phages (vB_SenA_SM5, Sh19. SE14, and vB-SalM-PM10) are proposed to have commercial utility in food-related industries, although none have been formally evaluated for cross-reactivity against C. sedlakii to our knowledge [ 61 , 62 , 63 , 64 ]. The procedures demonstrated in this study may thus have value in improving the specificity of other phages currently in development.…”

Section: Discussionmentioning

confidence: 99%

Tailoring the Host Range of Ackermannviridae Bacteriophages through Chimeric Tailspike Proteins

Gil

Paulson

Brown

et al. 2023

Viruses

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Host range is a major determinant in the industrial utility of a bacteriophage. A model host range permits broad recognition across serovars of a target bacterium while avoiding cross-reactivity with commensal microbiota. Searching for a naturally occurring bacteriophage with ideal host ranges is challenging, time-consuming, and restrictive. To address this, SPTD1.NL, a previously published luciferase reporter bacteriophage for Salmonella, was used to investigate manipulation of host range through receptor-binding protein engineering. Similar to related members of the Ackermannviridae bacteriophage family, SPTD1.NL possessed a receptor-binding protein gene cluster encoding four tailspike proteins, TSP1-4. Investigation of the native gene cluster through chimeric proteins identified TSP3 as the tailspike protein responsible for Salmonella detection. Further analysis of chimeric phages revealed that TSP2 contributed off-target Citrobacter recognition, whereas TSP1 and TSP4 were not essential for activity against any known host. To improve the host range of SPTD1.NL, TSP1 and TSP2 were sequentially replaced with chimeric receptor-binding proteins targeting Salmonella. This engineered construct, called RBP-SPTD1-3, was a superior diagnostic reporter, sensitively detecting additional Salmonella serovars while also demonstrating improved specificity. For industrial applications, bacteriophages of the Ackermannviridae family are thus uniquely versatile and may be engineered with multiple chimeric receptor-binding proteins to achieve a custom-tailored host range.

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Section: Discussionmentioning

confidence: 99%

Tailoring the Host Range of Ackermannviridae Bacteriophages through Chimeric Tailspike Proteins

Gil

Paulson

Brown

et al. 2023

Viruses

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“…The previous step was repeated thrice to obtain isolated phage plaques. All plates with no bacterial growth in the double-layer plaque assay were scrapped and mixed in sterile SM buffer [11]. The mixture was centrifuged, filtered, and incubated with the addition of 10% of polyethylene glycol 8000.…”

Section: Phage Isolation and Purificationmentioning

confidence: 99%

Genome Analysis and Antibiofilm Activity of Phage 590B against Multidrug-Resistant and Extensively Drug-Resistant Uropathogenic Escherichia coli Isolates, India

et al. 2022

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Urinary tract infections (UTIs) are among the most common bacterial infections in humans. Uropathogenic Escherichia coli (UPEC), which are the most frequent agents causing community as well as hospital-acquired UTIs, have become highly drug-resistant, thus making the treatment of these infections challenging. Recently, the use of bacteriophages (or ‘phages’) against multidrug-resistant (MDR) and extensively drug-resistant (XDR) microorganisms has garnered significant global attention. Bacterial biofilms play a vital role in the pathogenesis of UTIs caused by UPEC. Phages have the potential to disrupt bacterial biofilms using lytic enzymes such as EPS depolymerases and endolysins. We isolated a lytic phage (590B) from community sewage in Chandigarh, which was active against multiple MDR and XDR biofilm-forming UPEC strains. During whole-genome sequencing, the 44.3 kb long genome of phage 590B encoded 75 ORFs, of which 40 were functionally annotated based on homology with similar phage proteins in the database. Comparative analysis of associated phage genomes indicated that phage 590B evolved independently and had a distinct taxonomic position within the genus Kagunavirus in the subfamily Guernseyvirinae of Siphoviridae. The phage disrupted biofilm mass effectively when applied to 24 h old biofilms formed on the Foley silicon catheter and coverslip biofilm models. To study the effect of intact biofilm architecture on phage predation, the biofilms were disrupted. The phage reduced the viable cells by 0.6–1.0 order of magnitude after 24 h of incubation. Regrowth and intact bacterial cells were observed in the phage-treated planktonic culture and biofilms, respectively, which indicated the emergence of phage-resistant bacterial variants. The phage genome encoded an endolysin which might have a role in the disruption and inhibition of bacterial biofilms. Moreover, the genome lacked genes encoding toxins, virulence factors, antibiotic resistance, or lysogeny. Therefore, lytic phage 590B may be a good alternative to antibiotics and can be included in phage cocktails for the treatment of UTIs caused by biofilm-forming MDR and XDR UPEC strains.

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“…A single plaque was picked up via a sterile 1 mL micropipette tip, and this single phage was amplified, and all steps were repeated twice [13]. Ten percent of polyethylene glycol 8000 (PEG 8000, Sigma-Aldrich Corporation, Burlington, MA, USA) was added to the phage preparation, and incubated for 20 h at 4 • C. After centrifuging, the mixture at 257,000× g, the pellet was resuspended in the SM buffer (10 mM MgSO 4 •6H 2 O).…”

Section: Phage Purificationmentioning

confidence: 99%

Vibrio Phage VMJ710 Can Prevent and Treat Disease Caused by Pathogenic MDR V. cholerae O1 in an Infant Mouse Model

et al. 2023

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Cholera, a disease of antiquity, is still festering in developing countries that lack safe drinking water and sewage disposal. Vibrio cholerae, the causative agent of cholera, has developed multi-drug resistance to many antimicrobial agents. In aquatic habitats, phages are known to influence the occurrence and dispersion of pathogenic V. cholerae. We isolated Vibrio phage VMJ710 from a community sewage water sample of Manimajra, Chandigarh, in 2015 during an outbreak of cholera. It lysed 46% of multidrug-resistant V. cholerae O1 strains. It had significantly reduced the bacterial density within the first 4–6 h of treatment at the three multiplicity of infection (MOI 0.01, 0.1, and 1.0) values used. No bacterial resistance was observed against phage VMJ710 for 20 h in the time–kill assay. It is nearest to an ICP1 phage, i.e., Vibrio phage ICP1_2012 (MH310936.1), belonging to the class Caudoviricetes. ICP1 phages have been the dominant bacteriophages found in cholera patients’ stools since 2001. Comparative genome analysis of phage VMJ710 and related phages indicated a high level of genetic conservation. The phage was stable over a wide range of temperatures and pH, which will be an advantage for applications in different environmental settings. The phage VMJ710 showed a reduction in biofilm mass growth, bacterial dispersal, and a clear disruption of bacterial biofilm structure. We further tested the phage VMJ710 for its potential therapeutic and prophylactic properties using infant BALB/c mice. Bacterial counts were reduced significantly when phages were administered before and after the challenge of orogastric inoculation with V. cholerae serotype O1. A comprehensive whole genome study revealed no indication of lysogenic genes, genes associated with possible virulence factors, or antibiotic resistance. Based on all these properties, phage VMJ710 can be a suitable candidate for oral phage administration and could be a viable method of combatting cholera infection caused by MDR V. cholerae pathogenic strains.

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Complete Genome Sequence of Salmonella Phage vB_SenA_SM5, Active against Multidrug-Resistant Salmonella enterica Serovar Typhi Isolates

Cited by 3 publications

References 9 publications

Tailoring the Host Range of Ackermannviridae Bacteriophages through Chimeric Tailspike Proteins

Tailoring the Host Range of Ackermannviridae Bacteriophages through Chimeric Tailspike Proteins

Genome Analysis and Antibiofilm Activity of Phage 590B against Multidrug-Resistant and Extensively Drug-Resistant Uropathogenic Escherichia coli Isolates, India

Vibrio Phage VMJ710 Can Prevent and Treat Disease Caused by Pathogenic MDR V. cholerae O1 in an Infant Mouse Model

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