Genome Sequence of the Enterohemorrhagic Escherichia coli Bacteriophage UFV-AREG1

Lopez, Maryoris Elisa Soto; Batalha, Laís Silva; Vidigal, Pedro Marcus Pereira; Albino, Luiz Augusto Aguiar; Boggione, Delaine Meireles Gouveia; Gontijo, Marco Túlio Pardini; Bazzolli, Denise Mara Soares; Mendonça, Regina Célia Santos

doi:10.1128/genomea.00412-16

Cited by 14 publications

(7 citation statements)

References 6 publications

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“…Genome sequencing technology and metagenomics have dramatically expanded the understanding of phages and bacterial genomes, allowing screening for virulence, antibiotic-resistance genes [45,46] and antimicrobial peptides [18]. Alternative strategies to combat MDR bacteria include the use of lactic acid bacteria, probiotics and their bacteriocins [47][48][49], phages [50][51][52][53], phage-derived lytic enzymes and other antimicrobial proteins [54][55][56], as well as plant-based active molecules [57].…”

Section: Discussionmentioning

confidence: 99%

Combined effect of SAR-endolysin LysKpV475 with polymyxin B and Salmonella bacteriophage phSE-5

Gontijo,

Pereira Teles,

Martins Correia

et al. 2024

Microbiology

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Endolysins are bacteriophage (or phage)-encoded enzymes that catalyse the peptidoglycan breakdown in the bacterial cell wall. The exogenous action of recombinant phage endolysins against Gram-positive organisms has been extensively studied. However, the outer membrane acts as a physical barrier when considering the use of recombinant endolysins to combat Gram-negative bacteria. This study aimed to evaluate the antimicrobial activity of the SAR-endolysin LysKpV475 against Gram-negative bacteria as single or combined therapies, using an outer membrane permeabilizer (polymyxin B) and a phage, free or immobilized in a pullulan matrix. In the first step, the endolysin LysKpV475 in solution, alone and combined with polymyxin B, was tested in vitro and in vivo against ten Gram-negative bacteria, including highly virulent strains and multidrug-resistant isolates. In the second step, the lyophilized LysKpV475 endolysin was combined with the phage phSE-5 and investigated, free or immobilized in a pullulan matrix, against Salmonella enterica subsp. enterica serovar Typhimurium ATCC 13311. The bacteriostatic action of purified LysKpV475 varied between 8.125 μg ml−1 against Pseudomonas aeruginosa ATCC 27853, 16.25 μg ml−1 against S. enterica Typhimurium ATCC 13311, and 32.50 μg ml−1 against Klebsiella pneumoniae ATCC BAA-2146 and Enterobacter cloacae P2224. LysKpV475 showed bactericidal activity only for P. aeruginosa ATCC 27853 (32.50 μg ml−1) and P. aeruginosa P2307 (65.00 μg ml−1) at the tested concentrations. The effect of the LysKpV475 combined with polymyxin B increased against K. pneumoniae ATCC BAA-2146 [fractional inhibitory concentration index (FICI) 0.34; a value lower than 1.0 indicates an additive/combined effect] and S. enterica Typhimurium ATCC 13311 (FICI 0.93). A synergistic effect against S. enterica Typhimurium was also observed when the lyophilized LysKpV475 at ⅔ MIC was combined with the phage phSE-5 (m.o.i. of 100). The lyophilized LysKpV475 immobilized in a pullulan matrix maintained a significant Salmonella reduction of 2 logs after 6 h of treatment. These results demonstrate the potential of SAR-endolysins, alone or in combination with other treatments, in the free form or immobilized in solid matrices, which paves the way for their application in different areas, such as in biocontrol at the food processing stage, biosanitation of food contact surfaces and biopreservation of processed food in active food packing.

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Section: Discussionmentioning

confidence: 99%

Combined effect of SAR-endolysin LysKpV475 with polymyxin B and Salmonella bacteriophage phSE-5

Gontijo,

Pereira Teles,

Martins Correia

et al. 2024

Microbiology

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“…The strain was cultivated in Brain Heart Infusion Broth (HiMedia, Brazil) growth medium and incubated at 37°C. Bacteriophage UFV-AREG1 from Tequatrovirus ufvareg1 specie was previously isolated and sequenced (Lopez et al, 2016). Bacteriophages were stocked and diluted in SM buffer (gelatin 0,01% [m/v] [Vetec, Brazil], Tris-HCl 50 mM [pH 7,5] [Sigma Aldrich, Brazil], NaCl 100 mM [Vetec, Brazil], MgSO 4 .7H 2 O 8 mM [Vetec, Brazil]).…”

Section: Bacterial Strains and Culture Conditionsmentioning

confidence: 99%

Complete genome analysis of Tequatrovirus ufvareg1, a Tequatrovirus species inhibiting Escherichia coli O157:H7

Lopez

Gontijo

Cardoso

et al. 2023

Front. Cell. Infect. Microbiol.

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IntroductionBacteriophages infecting human pathogens have been considered potential biocontrol agents, and studying their genetic content is essential to their safe use in the food industry. Tequatrovirus ufvareg1 is a bacteriophage named UFV-AREG1, isolated from cowshed wastewater and previously tested for its ability to inhibit Escherichia coli O157:H7.MethodsT. ufvareg1 was previously isolated using E. coli O157:H7 (ATCC 43895) as a bacterial host. The same strain was used for bacteriophage propagation and the one-step growth curve. The genome of the T. ufvareg1 was sequenced using 305 Illumina HiSeq, and the genome comparison was calculated by VIRIDIC and VIPTree.ResultsHere, we characterize its genome and compare it to other Tequatrovirus. T. ufvareg1 virions have an icosahedral head (114 x 86 nm) and a contracted tail (117 x 23 nm), with a latent period of 25 min, and an average burst size was 18 phage particles per infected E. coli cell. The genome of the bacteriophage T. ufvareg1 contains 268 coding DNA sequences (CDS) and ten tRNA genes distributed in both negative and positive strains. T. ufvareg1 genome also contains 40 promoters on its regulatory regions and two rho-independent terminators. T. ufvareg1 shares an average intergenomic similarity (VIRIDC) of 88.77% and an average genomic similarity score (VipTree) of 88.91% with eight four reference genomes for Tequatrovirus available in the NCBI RefSeq database. The pan-genomic analysis confirmed the high conservation of Tequatrovirus genomes. Among all CDS annotated in the T. ufvareg1 genome, there are 123 core genes, 38 softcore genes, 94 shell genes, and 13 cloud genes. None of 268 CDS was classified as being exclusive of T. ufvareg1.ConclusionThe results in this paper, combined with other previously published findings, indicate that T. ufvareg1 bacteriophage is a potential candidate for food protection against E. coli O157:H7 in foods.

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“…Recently, use of phages against P. aeruginosa in the poultry industry was also evidenced in Minas Gerais State. 51 …”

Section: Brazilian Studies Using Phagesmentioning

confidence: 99%

Clinical utilization of bacteriophages: a new perspective to combat the antimicrobial resistance in Brazil

Martins

Toleman

Gales

2020

The Brazilian Journal of Infectious Diseases

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Genome Sequence of the Enterohemorrhagic Escherichia coli Bacteriophage UFV-AREG1

Abstract: Here, we present the genome sequence of the Escherichia coli bacteriophage UFV-AREG1. This phage was isolated from cowshed wastewater and showed specificity for enterohemorrhagic E. coli O157:H7 (ATCC 43895), E. coli 0111 (CDC O11ab) and E. coli (ATCC 23229).

Cited by 14 publications

References 6 publications

Combined effect of SAR-endolysin LysKpV475 with polymyxin B and Salmonella bacteriophage phSE-5

Combined effect of SAR-endolysin LysKpV475 with polymyxin B and Salmonella bacteriophage phSE-5

Complete genome analysis of Tequatrovirus ufvareg1, a Tequatrovirus species inhibiting Escherichia coli O157:H7

Clinical utilization of bacteriophages: a new perspective to combat the antimicrobial resistance in Brazil

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