Molecular detection of pathogens in water – The pros and cons of molecular techniques

Gironés, Rosina; Ferrús, María Antonia; Alonso, J.L.; Rodríguez-Manzano, Jesús; Calgua, Byron; Corrêa, Adriana de Abreu; Hundesa, Ayalkibet; Carratalà, Anna; Bofill-Mas, Sı́lvia

doi:10.1016/j.watres.2010.06.030

Cited by 371 publications

(273 citation statements)

References 128 publications

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“…It is known that the pollution and discharge of sewage alters the microbial community of freshwater environments (Paul and Meyer, 2001;Walsh et al, 2005;Girones et al, 2010). However, there is still a need to establish biological standards that can evaluate the water quality.…”

Section: Discussionmentioning

confidence: 99%

“…Most of those pathogens can be removed from the water by a sewage treatment but, when no treatment is performed, these pathogens can be discharged into effluents and can be a threat to public health (Girones et al, 2010). In Brazil, non-treated sewage is a major cause of water pollution.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, urbanisation can be a source of pathogenic bacteria through the discharge of human sewage into water environments (Girones et al, 2010). Through the monitoring of microbes in water, it is possible to identify potential pathogens.…”

Section: Introductionmentioning

confidence: 99%

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Characterization of the microbial community in a lotic environment to assess the effect of pollution on nitrifying and potentially pathogenic bacteria

et al. 2014

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This study aimed to investigate microbes involved in the nitrogen cycle and potentially pathogenic bacteria from urban and rural sites of the São Pedro stream. Water samples were collected from two sites. A seasonal survey of bacterial abundance was conducted. The dissolved nutrient content was analysed. PCR and FISH analysis were performed to identify and quantify microbes involved in the nitrogen cycle and potentially pathogenic bacteria. The seasonal survey revealed that the bacterial abundance was similar along the year on the rural area but varied on the urban site. Higher concentration of dissolved nutrients in the urban area indicated a eutrophic system. Considering the nitrifying microbes, the genus Nitrobacter was found, especially in the urban area, and may act as the principal bacteria in converting nitrite into nitrate at this site. The molecular markers napA, amoA, and nfrA were more accumulated at the urban site, justifying the higher content of nutrients metabolised by these enzymes. Finally, high intensity of amplicons from Enterococcus, Streptococcus, Bacteroides/Prevotella/Porphyromonas, Salmonella, S. aureus, P. aeruginosa and the diarrheagenic lineages of E. coli were observed at the urban site. These results indicate a change in the structure of the microbial community imposed by anthrophic actions. The incidence of pathogenic bacteria in aquatic environments is of particular importance to public health, emphasising the need for sewage treatment to minimise the environmental impacts associated with urbanisation.Keywords: lotic environment, urbanisation, pollution, nitrifying microbes, pathogenic bacteria.Caracterização da comunidade microbiana em um ambiente lótico para acessar o efeito da poluição em bactérias nitrificantes e potencialmente patogênicas ResumoEste estudo objetivou investigar os micro-organismos envolvidos no ciclo do nitrogênio e bactérias potencialmente patogênicas das áreas urbanas e rurais do Córrego São Pedro. Amostras de água foram coletadas dos dois locais. Um levantamento sazonal da densidade bacteriana foi realizado. O teor de nutriente dissolvido foi avaliado. As técnicas de PCR e FISH foram realizadas para identificar e quantificar os micro-organismos envolvidos no ciclo do nitrogênio e bactérias potencialmente patogênicas. O levantamento sazonal revelou que a abundância bacteriana foi semelhante ao longo do ano na área rural, porém variou na região urbana. Altas concentrações de nutrientes dissolvidos na área urbana indicaram este como um sistema eutrófico. Considerando os micro-organismos nitrificantes, o gênero Nitrobacter foi encontrado, especialmente na região urbana, e pode estar atuando como a principal bactéria convertendo nitrito em nitrato nessa área. Os marcadores moleculares napA, amoA, e nfrA foram mais acumulados na área urbana, justificando o alto teor dos nutrientes metabolizados por essas enzimas. Finalmente, alta intensidade de amplicons para Enterococcus, Streptococcus, Bacteroides/Prevotella/Porphyromonas, Salmonella, S. aureus, P. aerug...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Characterization of the microbial community in a lotic environment to assess the effect of pollution on nitrifying and potentially pathogenic bacteria

et al. 2014

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“…1 b) shows that the enzyme catalyzed reaction follows a pseudo-first order reaction kinetics with a simulated reaction rate constant of (7.361±0.633)10 3 s 1 (r 2 = 0.9713), illustrating that the enzyme's Michaelis constant (K M ) is much larger than the initial concentration of AA (100 µM), in accordance with previous publication. [24] The immobilized enzyme has a high local concentration, leading to fast enzyme-catalyzed AA oxidation. The A c c e p t e d M a n u s c r i p t 9 confined space of the microfluidic channel also contributes to the high efficiency of the reaction.…”

Section: On-chip Ascorbic Acid Oxidationmentioning

confidence: 99%

Interference-blind microfluidic sensor for ascorbic acid determination by UV/vis spectroscopy

Fernandes

Cardoso

et al. 2016

Sensors and Actuators B: Chemical

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A microfluidic sensor is developed and targeted at specific ingredients determination in drug/food/beverage matrices. The surface of a serpentine polydimethylsiloxane (PDMS) microchannel is modified by enzyme via physisorption. When solutions containing target ingredients pass through the microfluidic channel, enzyme-catalyzed reaction occurs and only converts the target molecules to its products. The whole process is monitored by an end-channel UV/vis spectroscopic detection. Ascorbate oxidase and L-ascorbic acid (AA) are taken as enzyme-substrate model in this study to investigate the feasibility of using the developed strategy for direct quantification of AA in standard solutions and complex matrices. A dietary supplement product, vitamin C tablet, is chosen as a model matrix to test the microfluidic bio-sensor in real-sample analysis. The results illustrate that the established microfluidic biosensor exhibits good reproducibility, stability, and anti-interference property. Technically, it is easy to realize, depends on low investment in chip fabrication, and simple instrumental procedure, where only UV/vis spectrophotometer is required. To sum up, the developed strategy is economical, specific, and accurate, and can be potentially used for fast quantification of ingredient in samples with complex matrix background. It is promising to be widely spread in food industry and quality control department.

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“…This rigorous detection scheme is not amenable to the rapid and high-volume screening of multiple environmental samples. Because of these shortcomings, a large body of research has been dedicated to finding rapid and high-throughput screening techniques for the presence of disease-causing organisms in different samples (Girones et al 2010;Noble and Weisberg 2005;Straub et al 2005). One promising and rapidly evolving avenue is biosensors capable of interacting with biological molecules at the nanometer scale (Vikesland and Wigginton 2010).…”

Section: Introductionmentioning

confidence: 99%

Rapid MPN-Qpcr Screening for Pathogens in Air, Soil, Water, and Agricultural Produce

Orlofsky

Benami

Gross

et al. 2015

Water Air Soil Pollut

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A sensitive, high-throughput, and costeffective method for screening bacterial pathogens in the environment was developed. A variety of environmental samples, including aerosols, soil of various types (sand, sand/clay mix, and clay), wastewater, and vegetable surface (modeled by tomato), were concomitantly spiked with Salmonella enterica and/or Pseudomonas aeruginosa to determine recovery rates and limits of detection. The various matrices were first enriched with a general pre-enrichment broth in a dilution series and then enumerated by most probable number (MPN) estimation using quantitative PCR for rapid screening of amplicon presence. Soil and aerosols were then tested in non-spiked environmental samples, as these matrices are prone to large experimental variation. Limit of detection in the various soil types was 1-3 colony-forming units (CFU) g −1 ; on vegetable surface, 5 CFU per tomato; in treated wastewater, 5 CFU L −1 ; and in aerosols, >300 CFU mL −1 . Our method accurately identified S. enterica in non-spiked environmental soil samples within a day, while traditional methods took 4 to 5 days and required sorting through biochemically and morphologically similar species. Likewise, our method successfully identified P. aeruginosa in non-spiked aerosols generated by a domestic wastewater treatment system. The obtained results suggest that the developed method presents a broad approach for the rapid, efficient, and reliable detection of relatively low densities of pathogenic organisms in challenging environmental samples.

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Molecular detection of pathogens in water – The pros and cons of molecular techniques

Cited by 371 publications

References 128 publications

Characterization of the microbial community in a lotic environment to assess the effect of pollution on nitrifying and potentially pathogenic bacteria

Characterization of the microbial community in a lotic environment to assess the effect of pollution on nitrifying and potentially pathogenic bacteria

Interference-blind microfluidic sensor for ascorbic acid determination by UV/vis spectroscopy

Rapid MPN-Qpcr Screening for Pathogens in Air, Soil, Water, and Agricultural Produce

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