Bluephage: A rapid method for the detection of somatic coliphages used as indicators of fecal pollution in water

Muniesa, Maite; Ballesté, Elisenda; Imamovic, Lejla; Pascual-Benito, Míriam; Toribio-Avedillo, Daniel; Lucena, F.; Blanch, Anicet R.; Jofre, Juan

doi:10.1016/j.watres.2017.10.030

Cited by 29 publications

(17 citation statements)

References 27 publications

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“…These methods are mostly characterized by the use of certain bacterial host strains. Recently, some fast and friendly methods that can be adapted to ready-to-use kits have become available (Salter et al 2010;Muniesa et al 2018). Molecular methods based on counting phage particles are on hand for some specific bacteriophages, mostly F-specific phages, but not for the entire operational groups (Wolf et al 2010).…”

Section: Viral Indicatorsmentioning

confidence: 99%

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Pathogens, faecal indicators and human-specific microbial source-tracking markers in sewage

et al. 2019

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Summary The objective of this review is to assess the current state of knowledge of pathogens, general faecal indicators and human‐specific microbial source tracking markers in sewage. Most of the microbes present in sewage are from the microbiota of the human gut, including pathogens. Bacteria and viruses are the most abundant groups of microbes in the human gut microbiota. Most reports on this topic show that raw sewage microbiological profiles reflect the human gut microbiota. Human and animal faeces share many commensal microbes as well as pathogens. Faecal‐orally transmitted pathogens constitute a serious public health problem that can be minimized through sanitation. Assessing both the sanitation processes and the contribution of sewage to the faecal contamination of water bodies requires knowledge of the content of pathogens in sewage, microbes indicating general faecal contamination and microbes that are only present in human faecal remains, which are known as the human‐specific microbial source‐tracking (MST) markers. Detection of pathogens would be the ideal option for managing sanitation and determining the microbiological quality of waters contaminated by sewage; but at present, this is neither practical nor feasible in routine testing. Traditionally, faecal indicator bacteria have been used as surrogate indicators of general faecal residues. However, in many water management circumstances, it becomes necessary to detect both the origin of faecal contamination, for which MST is paramount, and live micro‐organisms, for which molecular methods are not suitable. The presence and concentrations of pathogens, general faecal indicators and human‐specific MST markers most frequently reported in different areas of the world are summarized in this review.

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Section: Viral Indicatorsmentioning

confidence: 99%

“…; Muniesa et al . ). Molecular methods based on counting phage particles are on hand for some specific bacteriophages, mostly F‐specific phages, but not for the entire operational groups (Wolf et al .…”

Section: Introductionmentioning

confidence: 97%

Pathogens, faecal indicators and human-specific microbial source-tracking markers in sewage

et al. 2019

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“…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 mL.The aim of this study was to compare the accuracy of the Bluephage method in testing for the presence/ absence of somatic coliphages in 100 mL volumes of water, as well as to assess its specificity and detection limit compared to the reference method ISO-DAL.…”

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“…When ready, 10 mL of the culture was added to a 100 mL volume of water for testing. The culture was previously obtained using a dry culture medium (fast kit BPF-SPA, Bluephage S. L., Barcelona) containing the necessary compounds to obtain the exact broth composition as described by Muniesa et al 18 . The water sample receiving the CB10 culture was swirled gently until thoroughly mixed and incubated at (36 ± 1) °C.…”

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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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Modeling the photoinactivation and transport of somatic and F‐specific coliphages at a Great Lakes beach

et al. 2020

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Fecal indicator organisms (FIOs), such as Escherichia coli and enterococci, are often used as surrogates of contamination in the context of beach management; however, bacteriophages may be more reliable indicators than FIO due to their similarity to viral pathogens in terms of size and persistence in the environment. In the past, mechanistic modeling of environmental contamination has focused on FIOs, with virus and bacteriophage modeling efforts remaining limited. In this paper, we describe the development and application of a fate and transport model of somatic and F-specific coliphages for the Washington Park beach in Lake Michigan, which is affected by riverine outputs from the nearby Trail Creek. A three-dimensional model of coliphage transport and photoinactivation was tested and compared with a previously reported E. coli fate and transport model. The light-based inactivation of the phages was modeled using organism-specific action spectra. Results indicate that the coliphage models outperformed the E. coli model in terms of reliably predicting observed E. coli/coliphage concentrations at the beach. This is possibly due to the presence of additional E. coli sources that were not accounted for in the modeling. The coliphage models can be used to test hypotheses about potential sources and their behavior and for predictive modeling.

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Bluephage: A rapid method for the detection of somatic coliphages used as indicators of fecal pollution in water

Cited by 29 publications

References 27 publications

Pathogens, faecal indicators and human-specific microbial source-tracking markers in sewage

Pathogens, faecal indicators and human-specific microbial source-tracking markers in sewage

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

Modeling the photoinactivation and transport of somatic and F‐specific coliphages at a Great Lakes beach

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