Comparison of single and multi-host enrichment approach for harnessing lytic phages against antimicrobial-resistant E. coli: Repurposing the enrichment step

Vaiyapuri, Murugadas; Karthika, R.; George, Iris; Gundubilli, Devika; Visnuvinayagam, S.; Krishnan, S.; George, Joshy Chalil; Mothadaka, Mukteswar Prasad; Nagarajarao, Ravishankar Chandragiri; Badireddy, Madhusudana Rao

doi:10.2478/s11756-020-00652-2

Cited by 10 publications

(3 citation statements)

References 48 publications

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“…Isolation of bacteriophages was performed by multiple host enrichment method with modification in the selective media and incubation conditions (Vaiyapuri et al . 2021). Among the 50 LAB isolates obtained from dahi identified based on 16S rRNA gene sequencing and characterised based on technological properties, L. fermentum strains showed most of the desirable technological properties, including faster acidification, diacetyl production and ability to produce exopolysaccharides.…”

Section: Methodsmentioning

confidence: 99%

Selection of robust starters from household dahi and assessment of their sensitivity to bacteriophage isolated from dairy effluents

Chandran,

Vaiyapuri,

Rathish

et al. 2024

Int J of Dairy Tech

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Identifying the predominant strains of lactic acid bacteria (LAB) in dahi that are resistant to bacteriophages is essential for developing robust starter cultures. Dahi samples from two agroecological zones of Kerala were collected for isolation of LAB. Limosilactobacillus fermentum emerged as the dominant starter culture species based on sequence‐based identification and technological property characterisation. Dairy effluents were screened for phages targeting L. fermentum, and 10 of 11 strains of L. fermentum showed resistance to one lytic phage isolated as part of this study. The strain ADMT 15 showed sensitivity to the phage and on subsequent genome sequencing the phage revealed its close relationship to members of the Herelleviridae phage family.

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Section: Methodsmentioning

confidence: 99%

Selection of robust starters from household dahi and assessment of their sensitivity to bacteriophage isolated from dairy effluents

Chandran,

Vaiyapuri,

Rathish

et al. 2024

Int J of Dairy Tech

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“…The filtrate was then screened for the presence of bacteriophages by multiple host enrichment method with modification for Lc. paracasei host whose details are given in the Supplementary File (and see Vaiyapuri et al 2021).…”

Section: Phage Isolation and Enrichmentmentioning

confidence: 99%

Detection of lytic phage infecting flavour-producing strain of Lacticaseibacillus paracasei in the dairy effluents of Kerala

Chandran

Beena

Vaiyapuri

et al. 2023

Journal of Dairy Research

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The performance of the starter culture is a critical factor that decides the quality of fermented milk. Dahi is a fermented milk product popular in India made using a mixed starter culture of lactic acid bacteria comprising acid and flavour producers. The prevalence of bacteriophages in the dairy environment can critically affect the activity of these starter cultures resulting in starter failure. As there is little information available on the occurrence of bacteriophages in the dairy environment of Kerala, this research communication examines the presence of lytic bacteriophages acting against three potential flavour-producing strains of Lacticaseibacillus paracasei (Lc. paracasei). Dairy effluent samples were screened for the presence of phages against the strains of Lc. paracasei by the multiple host enrichment method. Plates showing clearance zone in spot assay were confirmed for the presence of phages by double-layer agar assay. The plaques obtained in the double-layer agar assay were purified for further identification by next-generation sequencing. A bacteriophage infecting one of the three strains of Lc. paracasei was detected by the plaque assay and the blast annotation of the bacteriophage sequence found 86.05% similarity of the phage to Siphoviridae family. The study endorses the need for monitoring phages in the dairy environment to control phage-related starter failure in the state of Kerala.

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“…The isolated plaques were picked, and the process was repeated three times to obtain purified vibriophage with consistent plaque morphology. Phage precipitation was performed by treating purified vibriophage (25 mL) with polyethylene glycol (10% w/v PEG 8000 and 1 M NaCl) at 4 • C for 1 h, followed by overnight incubation at −20 • C for 24 h and centrifugation at 10,000 rpm for 20 min at 4 • C [45,46]. The pellet was resuspended in SM buffer (100 mM NaCl, 8 mM MgSO 4 .7H 2 O, 50 mM Tris-Cl, pH 7.5), which constituted the purified and enriched vibriophage, and stored at 4 • C for downstream analysis.…”

Section: Purification and Precipitation Of Vibriophagementioning

confidence: 99%

Genome Characterization and Infectivity Potential of Vibriophage-ϕLV6 with Lytic Activity against Luminescent Vibrios of Penaeus vannamei Shrimp Aquaculture

Benala

Vaiyapuri

Visnuvinayagam

et al. 2023

Viruses

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Shrimp aquaculture, especially during the hatchery phase, is prone to economic losses due to infections caused by luminescent vibrios. In the wake of antimicrobial resistance (AMR) in bacteria and the food safety requirements of farmed shrimp, aqua culturists are seeking alternatives to antibiotics for shrimp health management, and bacteriophages are fast emerging as natural and bacteria-specific antimicrobial agents. This study analyzed the whole genome of vibriophage-ϕLV6 that showed lytic activity against six luminescent vibrios isolated from the larval tanks of P. vannamei shrimp hatcheries. The Vibriophage-ϕLV6 genome was 79,862 bp long with 48% G+C content and 107 ORFs that coded for 31 predicted protein functions, 75 hypothetical proteins, and a tRNA. Pertinently, the vibriophage-ϕLV6 genome harbored neither AMR determinants nor virulence genes, indicating its suitability for phage therapy. There is a paucity of whole genome-based information on vibriophages that lyse luminescent vibrios, and this study adds pertinent data to the database of V. harveyi infecting phage genomes and, to our knowledge, is the first vibriophage genome report from India. Transmission electron microscopy (TEM) of vibriophage-ϕLV6 revealed an icosahedral head (~73 nm) and a long, flexible tail (~191 nm) suggesting siphovirus morphology. The vibriophage-ϕLV6 phage at a multiplicity of infection (MOI) of 80 inhibited the growth of luminescent V. harveyi at 0.25%, 0.5%, 1%, 1.5%, 2%, 2.5%, and 3% salt gradients. In vivo experiments conducted with post-larvae of shrimp showed that vibriophage-ϕLV6 reduced luminescent vibrio counts and post-larval mortalities in the phage-treated tank compared to the bacteria-challenged tank, suggesting the potentiality of vibriophage-ϕLV6 as a promising candidate in treating luminescent vibriosis in shrimp aquaculture. The vibriophage-ϕLV6 survived for 30 days in salt (NaCl) concentrations ranging from 5 ppt to 50 ppt and was stable at 4 °C for 12 months.

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Comparison of single and multi-host enrichment approach for harnessing lytic phages against antimicrobial-resistant E. coli: Repurposing the enrichment step

Cited by 10 publications

References 48 publications

Selection of robust starters from household dahi and assessment of their sensitivity to bacteriophage isolated from dairy effluents

Selection of robust starters from household dahi and assessment of their sensitivity to bacteriophage isolated from dairy effluents

Detection of lytic phage infecting flavour-producing strain of Lacticaseibacillus paracasei in the dairy effluents of Kerala

Genome Characterization and Infectivity Potential of Vibriophage-ϕLV6 with Lytic Activity against Luminescent Vibrios of Penaeus vannamei Shrimp Aquaculture

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