Isolation and characterization of bacteriophages with lytic activity against common bacterial pathogens

Shende, Rajesh Kumar; Hirpurkar, S. D.; Sannat, Chandrahas; Rawat, Nidhi; Pandey, Vinay Kumar

doi:10.14202/vetworld.2017.973-978

Cited by 48 publications

(43 citation statements)

References 23 publications

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“…Both UPEC04 and UPEC10 were stable to heat up to 60℃. At 70℃, the phages were inactivated which is in agreement with Lu et al (2003) and Shende et al (2017) who reported that phages get inactivated at 70℃ and above [51,53].…”

Section: Discussionsupporting

confidence: 88%

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Bacteriophage activity against and characterisation of avian pathogenicEscherichia coliisolated from colibacillosis cases in Uganda

Kazibwe

Katami

Alinaitwe

et al. 2020

Preprint

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A laboratory-based study aimed at establishing a stock of avian pathogenic Escherichia coli (APEC) lytic bacteriophages, for future development of cocktail products for controlling colibacillosis as well as minimizing use of antimicrobial drugs in the poultry production systems in Uganda. Specifically, the study determined the antibiotic susceptibility; phylogenetic categories, occurrence of selected virulence genes among Escherichia coli stock isolates from cases of chicken colibacillosis; and isolation of specific bacteriophages. Fifty six isolates were confirmed as E. coli by standard phenotypic tests. All the 56 (100%) isolates were resistant to at least one antibiotic while 50 (89.3%) isolates were resistant to at least three classes of antimicrobial drugs and were therefore designated as multi-drug resistant. Phylogenetically, APEC isolates mainly belonged to phylogroups A and D which represented 44.6% and 39.3%, respectively. Virulence genes, ompT and iutA were the most frequent with 33 (58.9%) and 32 (57.1%) isolates respectively; while iroN least occurred in 23 (41.1%) isolates. Of the 56 isolates, 69.6% harbored at least one virulence gene, while 50% had at least four virulence genes; hence confirmed as APEC. None of the isolates belonged to the selected serotypes O1, O2 and O78. Seven specific bacteriophages were isolated and their host range, varied from 1.8% to 17.9% (n=56 APEC isolates), while the combined lytic spectrum of all the phages was 25%. Phage stability was negatively affected by increasing temperatures with both UPEC04 and UPEC10 phages becoming undetectable at 70°C; however activity was detected between pH 2 and 12. The high occurrence of APEC isolates with resistance against the commonly used antibiotics supports the need for alternative strategies of bacterial infections control in poultry. The low host range exhibited by the phages calls for search for more candidates before more in-depth studies are done for phage characterization and application.

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Section: Discussionsupporting

confidence: 88%

“…The main physical factors affecting phage adsorption and growth include pH and temperature [53]. The different pH and temperature ranges in this study were selected to mimic those that would be encountered during the handling and application of these phages as therapeutic or sanitizing bio-control agents on poultry farms.…”

Section: Discussionmentioning

confidence: 99%

Bacteriophage activity against and characterisation of avian pathogenicEscherichia coliisolated from colibacillosis cases in Uganda

Kazibwe

Katami

Alinaitwe

et al. 2020

Preprint

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“…De acordo com Shende et al (2017), fagos com amplitude de hospedeiro do tipo polivalente possuem potencial para substituir os antibióticos, e os fagosPedr3Sa, Pedr3Ec e Pedr3Pa apresentaram amplitude deste tipo, porém, deve ser considerado que a investigação de fagos com o intuito de Tabela 1. Características morfológicas do ensaio de placa.…”

Section: Resultsunclassified

Análise da amplitude de hospedeiros de vírus bacterianos isolados de amostras de esgoto doméstico da cidade de Macapá

Costa¹,

Neto²,

Souza³

et al. 2019

Revista Arquivos Científicos (IMMES)

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Palavras-chaveBacteriófagos Agentes anti-infecciosos Antimicrobiano Bacteriófagos ou fagos, são vírus que infectam e matam bactérias com grande seletividade, por isso, possuem grande aplicabilidade terapêutica e biotecnológica para o controle de bactérias de forma abrangente ou especifica em função de sua amplitude de hospedeiros, ou seja, a capacidade de infectar e lisar diferentes espécies bacterianas. O que os potencializa como uma poderosa ferramenta para o biocontrole bacteriano, incluindo patógenos, principalmente em um contexto na qual a resistência bacteriana aos antibióticos ameaça a eficácia desses fármacos e gera muitos impactos negativos sobre a saúde pública ao nível global. Fato este que torna relevante a realização de pesquisas com o intuito de isolar e explorar o potencial biotecnológico/terapêutico destes vírus como alternativa aos antibióticos. Neste contexto, este trabalho teve como objetivo isolar e caracterizar a amplitude de hospedeiros de bacteriófagos líticos provenientes de amostras de um esgoto da cidade de Macapá por meio de ensaios clássicos de microbiologia, que tiveram como resultado o isolamento de 4 bacteriófagos líticos, das quais 3 apresentaram amplitude de hospedeiro polivalente e 1 amplitude de hospedeiro do tipo monovalente. Keywords Bacteriophages Anti-infectives AntimicrobialBacteriophages, or phages, are viruses that infect and kill bacteria with high selectivity, so they have great therapeutic and biotechnological applicability for the control of bacteria in a wide or specific way depending on their host range, ie the ability to infect and lyse different bacterial species. This potentializes them as a powerful tool for bacterial biocontrol, including pathogens, especially in a context in which the bacterial resistance to the antibiotics threatens the efficacy of these drugs and generates many negative impacts over the public health globally. This fact makes research relevant to isolate and exploit the biotechnological/therapeutic potential of these viruses as an alternative to antibiotics. In this context, this work aimed to isolate and characterize the host range of lytic bacteriophage isolated from samples of sewage from the city of Macapá by classical microbiology assays, which resulted in the isolation of 4 lytic bacteriophages, of which 3 presented polyvalent host amplitude and 1 monovalent host amplitude.

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“…One-step growth experiments were carried out by a modification of methods described elsewhere [27,28].The logarithmic phase mixed with an overdose of phage host (MOI 10), 37 ℃ warm bath after 15 min 12000 rpm centrifugal 1 min, discard supernatant and wash with LB to precipitate .Then, suspended in LB that had been pre-warmed to 37 °C, followed by incubation at 37 °C. Samples were taken at 10 min intervals (up to 90min) and immediately diluted, and phage titers were then determined by double-layered agar plate method.…”

Section: One-step Growthmentioning

confidence: 99%

Characterization and genome analysis of novel phage vB_KpnM _Bp5 infecting Klebsiella pneumonia

Zhang

Wei

et al. 2019

Preprint

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Background With the incidence of antibiotic resistance reaching crisis point, it is imperative to find alternative treatments for multidrug-resistant infections. Using phage for pathogen control might be a promising treatment option to combat bacterial resistance. Results In this study, a lytic phage, designated vB_KpnM _Bp5, was isolated from pig faecal sample in Nanning, Guangxi province of China, and classified as a member of the family Muscle virus based on electron microscopy analysis. A one-step growth curve of the phage at the optimal MOI revealed that the latent time was 40 min and the burst size was 24 PFU/cell, indicative of good lysis capacity. Whole genome sequencing showed that phage vB_KpnM _Bp5 had a small dsDNA genome of 43872 bp. BLASTn analysis showed that it shared 94.06% identity (94% genome coverage) with Klebsiella phage vB_KpnP_SU552A of complete genome idefix. RAST genome analysis showed that the phage had 50 ORFs due to its small genome size, and the number of functional proteins was consistent with other phages. To evaluate the therapeutic effect of Klebsiella pneumoniae infection in mice, the results showed that phages provided vB_KpnM _Bp5better protection. Conclusion The phage vB_KpnM _Bp5 had the characteristics of broad host spectrum, strong environmental adaptability, short incubation period, large outbreak amount, and can cure the mouse model infected by Klebsiella pneumoniae. These findings suggested that phage vB_KpnM _Bp5 could be considered a potential therapeutic or prophylactic candidate against Klebsiella pneumonia infection.

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Isolation and characterization of bacteriophages with lytic activity against common bacterial pathogens

Cited by 48 publications

References 23 publications

Bacteriophage activity against and characterisation of avian pathogenicEscherichia coliisolated from colibacillosis cases in Uganda

Bacteriophage activity against and characterisation of avian pathogenicEscherichia coliisolated from colibacillosis cases in Uganda

Análise da amplitude de hospedeiros de vírus bacterianos isolados de amostras de esgoto doméstico da cidade de Macapá

Characterization and genome analysis of novel phage vB_KpnM _Bp5 infecting Klebsiella pneumonia

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