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Muniesa, Maite; Mocé-Llivina, Laura; Katayama, Hiroyuki; Jofre, Juan

doi:10.1023/a:1023384714481

Cited by 37 publications

(7 citation statements)

References 25 publications

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“…To elucidate this point, a set of experiments aiming to transduce the genes from phage particles isolated from the samples in E. coli was attempted. Unfortunately, as shown in other studies [42], this approach might need to identify a suitable and sensitive host strain ( E. coli or others) that would support infection with these phages and subsequent transduction. The search for the suitable host and the right conditions for transduction to occur is likely a complicated task.…”

Section: Discussionmentioning

confidence: 99%

Antibiotic Resistance Genes in the Bacteriophage DNA Fraction of Environmental Samples

2011

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Antibiotic resistance is an increasing global problem resulting from the pressure of antibiotic usage, greater mobility of the population, and industrialization. Many antibiotic resistance genes are believed to have originated in microorganisms in the environment, and to have been transferred to other bacteria through mobile genetic elements. Among others, β-lactam antibiotics show clinical efficacy and low toxicity, and they are thus widely used as antimicrobials. Resistance to β-lactam antibiotics is conferred by β-lactamase genes and penicillin-binding proteins, which are chromosomal- or plasmid-encoded, although there is little information available on the contribution of other mobile genetic elements, such as phages. This study is focused on three genes that confer resistance to β-lactam antibiotics, namely two β-lactamase genes (blaTEM and blaCTX-M9) and one encoding a penicillin-binding protein (mecA) in bacteriophage DNA isolated from environmental water samples. The three genes were quantified in the DNA isolated from bacteriophages collected from 30 urban sewage and river water samples, using quantitative PCR amplification. All three genes were detected in the DNA of phages from all the samples tested, in some cases reaching 104 gene copies (GC) of blaTEM or 102 GC of blaCTX-M and mecA. These values are consistent with the amount of fecal pollution in the sample, except for mecA, which showed a higher number of copies in river water samples than in urban sewage. The bla genes from phage DNA were transferred by electroporation to sensitive host bacteria, which became resistant to ampicillin. blaTEM and blaCTX were detected in the DNA of the resistant clones after transfection. This study indicates that phages are reservoirs of resistance genes in the environment.

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Section: Discussionmentioning

confidence: 99%

Antibiotic Resistance Genes in the Bacteriophage DNA Fraction of Environmental Samples

2011

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“…culture collections and others isolated by us and characterized by electron microscopy 20 . The spot test performed according to ISO 7 was carried out with bacteriophages belonging to different families of somatic bacteriophages: Microviridae (Phi X174 and M4), Myoviridae (SOM 1, SOM 3, SOM 5, SOM 8, SOM 15, SCH2, SCH10 and STER 5), Podoviridae (933 W), Siphoviridae (SOM 4, SOM7, SOM 28, SSAU 9, MAR 1 and SCH4) and Tectiviridae (PDR1).…”

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confidence: 99%

Bluephage, a method for efficient detection of somatic coliphages in one hundred milliliter water samples

Méndez

Toribio-Avedillo

Mangas-Casas

et al. 2020

Sci Rep

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Emerging water quality guidelines and regulations require the absence of somatic coliphages in 100 mL of water, yet the efficiency of standardized methods to test this volume of sample is questionable. A recently described procedure, Bluephage, using a modified E. coli host strain, overcomes some of the methodological limitations of standardized methods. In a maximum of 6.5 hours (2.5 hours for pre-growing the host strain and 4 hours for the presence/absence test), Bluephage allows the direct detection of one plaque-forming unit (PFU) in a 100 mL water sample. The test shows high levels of specificity for somatic coliphages and comparable accuracy with standardized methods.Coliphages, viruses infecting Escherichia coli, have been included or are being considered for inclusion in water quality regulations and guidelines around the world 1-6 . Some of these regulations, such as those for different types of drinking water 5,6 , prescribe or recommend the absence of somatic coliphages in 100 mL water samples.Widely accepted standardized methods for determining somatic coliphages are available 7-9 . The host strains and media endorsed in the ISO (International Organization for Standardization) standard 10705-2:2000 7 and the USEPA (U.S. Environmental Protection Agency) 1601 and 1602 methods 8,9 provide similar results 10-12 . To determine coliphage absence in 100 mL water samples, which requires analysis of the full 100 mL volume, the following standardized procedures are currently used: (1) the presence/ absence test according to ISO or USEPA; (2) the double agar layer (DAL) assay as indicated in ISO 7 , which requires the concentration of phages in 100 mL of water sample followed by phage enumeration in the full concentrate, and (3) the USEPA's single agar layer (SAL) assay 8 , which is applied to 10 replicas of 10 mL. According to ISO and USEPA standards, the presence/absence test requires 2 steps (enrichment and spot test) and needs more than one working day to obtain results. On the other hand, both the concentration 13,14 and SAL procedures are complex and suffer from loss of efficiency in coliphage recovery 10,15 . Feasible, efficient, fast and user-friendly methods to determine the presence of coliphages in 100 mL water samples are therefore required. Recent progress in this field [16][17][18] includes the development of Bluephage method 18 , whose application can facilitate the routine implementation of somatic coliphage determination in laboratories with minimal equipment and expertise requirements.To detect phage concentrations as low as one in a given volume is challenging, as viral particles have an uneven spatial distribution in watery suspensions, theoretically following the Poisson model 19 . Accordingly, to assess the accuracy and detection limit of different phage determination methods, a statistical approach was used to calculate the distribution in aliquots of suspension and consequently the expected percentage of positive samples in a set of recipients prepared to contain about one phage per 100 m...

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“…Somatic coliphages are a heterogeneous group of bacteriophages capable of infecting E. coli and other coliform bacteria through the cell wall after becoming attached to specific receptors on the outer membrane [12]. Under optimal physiological conditions, lysis occurs approximately 30 min after attachment, and between 100 and 1000 of progeny are released per infected cell [13].…”

Section: Families Of Bacteriophages Used As Indicators Of Fecal Pollu...mentioning

confidence: 99%

Bacteriophages as Fecal Pollution Indicators

Toribio-Avedillo

Blanch

Muniesa

et al. 2021

Viruses

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Bacteriophages are promising tools for the detection of fecal pollution in different environments, and particularly for viral pathogen risk assessment. Having similar morphological and biological characteristics, bacteriophages mimic the fate and transport of enteric viruses. Enteric bacteriophages, especially phages infecting Escherichia coli (coliphages), have been proposed as alternatives or complements to fecal indicator bacteria. Here, we provide a general overview of the potential use of enteric bacteriophages as fecal and viral indicators in different environments, as well as the available methods for their detection and enumeration, and the regulations for their application.

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Cited by 37 publications

References 25 publications

Antibiotic Resistance Genes in the Bacteriophage DNA Fraction of Environmental Samples

Antibiotic Resistance Genes in the Bacteriophage DNA Fraction of Environmental Samples

Bluephage, a method for efficient detection of somatic coliphages in one hundred milliliter water samples

Bacteriophages as Fecal Pollution Indicators

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