Population and single cell metabolic activity of UV-induced VBNC bacteria determined by CTC-FCM and D2O-labeled Raman spectroscopy

Guo, Lizheng; Ye, Chengsong; Li, Cui; Wan, Kun; Chen, Sheng; Zhang, Shenghua; Yu, Xin

doi:10.1016/j.envint.2019.05.077

Cited by 75 publications

(57 citation statements)

References 50 publications

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“…The validity of these tests to assess disinfection of drinking water, however, is debatable, as the majority of microorganisms in drinking water cannot currently be cultivated 16 , bacteria in drinking water are diverse, and bacteria in the environment have an increased UV resistance compared with laboratory-cultivated strains 1 . Exposure to UV can also cause some bacteria to enter a viable but nonculturable state as a response to environmental stress 17,18 .…”

Section: Introductionmentioning

confidence: 99%

Impact of UV irradiation at full scale on bacterial communities in drinking water

Pullerits

Ahlinder

Holmer³

et al. 2020

npj Clean Water

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Water in a full-scale drinking water treatment plant was irradiated with ultraviolet (UV) doses of 250, 400, and 600 J/m 2 , and the effect on bacterial communities investigated using 16s rRNA gene amplicon sequencing, heterotrophic plate counts (HPCs), coliform, and Escherichia coli counts. The bacteria in the irradiated water were also analyzed following storage for 6 days at 7°C, to approximate the conditions in the distribution system. The log 10 reduction of HPCs at 400 J/m 2 was 0.43 ± 0.12. Phylogenetic examination, including DESeq2 analysis, showed that Actinobacteria was more resistant to UV irradiation, whereas Bacteroidetes was sensitive to UV. Phylum Proteobacteria contained monophyletic groups that were either sensitive or resistant to UV exposure. The amplicon sequence variants (ASVs) resistant to UV irradiation had a greater average GC content than the ASVs sensitive to UV, at 55% ± 1.7 (n = 19) and 49% ± 2.5 (n = 16), respectively. Families Chitinophagaceae, Pelagibacteraceae, Holophagaceae, Methylophilaceae, and Cytophagaceae decreased linearly in relative abundance, with increasing UV dose (P < 0.05, Pearson's correlation). When irradiated water was stored, Chitinophagaceae, Comamonadaceae, and Flavobacteriaceae families decreased in relative abundance, whereas ACK-M1, Mycobacteriaceae, and Nitrosomonadaceae were increasing in relative abundance. This suggests that the impact of UV irradiation cannot only be considered directly after application but that this treatment step likely continues to influence microbial dynamics throughout the distribution system.npj Clean Water (2020) 3:11 ; https://doi.

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Section: Introductionmentioning

confidence: 99%

Impact of UV irradiation at full scale on bacterial communities in drinking water

Pullerits

Ahlinder

Holmer³

et al. 2020

npj Clean Water

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“…In 1982, Xu et al (1982) firstly reported “viable but non-culturable” (VBNC) state, which was considered to be a survival strategy of non-spore-forming bacteria in response to adverse conditions ( Xu et al, 1982 ; Oliver, 2010 ). Bacteria can enter into VBNC state by the stimulation of adverse environmental conditions, such as low temperature, nutrient-limited conditions, high salt, low pH, and even ultraviolet-induced ( Foster, 1999 ; Ramaiah et al, 2002 ; Cunningham et al, 2009 ; Deng et al, 2015 ; Liu et al, 2017a,b , 2018a,b ; Guo et al, 2019 ). For now, 85 species of bacteria have been confirmed that can enter into VBNC state, including 18 non-pathogenic and 67 pathogenic species ( Li et al, 2014 ).…”

Section: Introductionmentioning

confidence: 99%

Development and Application of a Simple “Easy To Operate” Propidium Monoazide-Crossing Priming Amplification on Detection of Viable and Viable But Non-culturable Cells of O157 Escherichia coli

Zhou

Wang

et al. 2020

Front. Microbiol.

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O157 Escherichia coli is one of the most important foodborne pathogens causing disease even at low cellular numbers. Thus, the early and accurate detection of this pathogen is important. However, due to the formation of viable but non-culturable (VBNC) status, the golden standard culturing methodology fails to identify O157 E. coli once it enters VBNC status. Crossing priming amplification (CPA) is a novel, simple, easy-to-operate detection technology that amplifies DNA with high speed, efficiency, and specificity under isothermal conditions. The objective of this study was to firstly develop and apply a CPA assay with propidium monoazide (PMA) for the rapid detection of the foodborne E. coli O157:H7 in VBNC state. Five primers (2a/1s, 2a, 3a, 4s, and 5a) were specially designed for recognizing three targets, which were rfbE, stx1, and stx2, and evaluated for its effectiveness in detecting VBNC cell of E. coli O157:H7 with detection limits of pure VBNC culture at 10 3 , 10 5 , and 10 5 colony-forming units (CFUs)/ml for rfbE, stx1, and stx2, respectively, whereas those of food samples (frozen pastry and steamed bread) were 10 3 , 10 5 , and 10 5 CFUs/ml. The application of the PMA-CPA assay was successfully used on detecting E. coli O157:H7 in VBNC state from food samples. In conclusion, this is the first development of PMA-CPA assay on the detection of VBNC cell, which was found to be useful and a powerful tool for the rapid detection of E. coli O157:H7 in VBNC state. Undoubtedly, the PMA-CPA method can be of high value to the food industry owing to its various advantages such as speed, specificity, sensitivity, and cost-effectiveness.

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“…After norfloxacin treatment, the number of VBNC bacterial cells was determined as the difference between the viable cell counts and the culturable cells counts using CTC and flow cytometry and the standard plate count methods ( Guo et al, 2019 ). As shown in Figure 1A , the total cells counts remained 10 8 cells/mL, while the number of living cells and cultured cells decreased gradually.…”

Section: Resultsmentioning

confidence: 99%

Alterations in the Cell Wall of Rhodococcus biphenylivorans Under Norfloxacin Stress

Jia

Fan

et al. 2020

Front. Microbiol.

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Many microorganisms can enter a viable but non-culturable (VBNC) state under various environmental stresses, while they can also resuscitate when the surroundings turn to suitable conditions. Cell walls play a vital role in maintaining cellular integrity and protecting cells from ambient threats. Here, we investigated the alterations in the cell wall of Rhodococcus biphenylivorans TG9 at VBNC state under norfloxacin stress and then at resuscitated state in fresh lysogeny broth medium. Electron microscopy analyses presented that TG9 in the VBNC state had a thicker and rougher cell wall than that in exponential phase or resuscitated state. Meanwhile, the results from infrared spectroscopy also showed that its VBNC state has different peptidoglycan structures in the cell wall. Moreover, in the VBNC cells the gene expressions related to cell wall synthesis and remodeling maintain a relatively high level. It indicates that the morphological variations of TG9 at the VBNC state might result from kinetic changes in the cell wall synthesis and remodeling. As a consequence, the alterations in the cell wall of VBNC TG9 may somewhat account for its tolerance mechanisms to antibiotic treatment.

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Population and single cell metabolic activity of UV-induced VBNC bacteria determined by CTC-FCM and D2O-labeled Raman spectroscopy

Cited by 75 publications

References 50 publications

Impact of UV irradiation at full scale on bacterial communities in drinking water

Impact of UV irradiation at full scale on bacterial communities in drinking water

Development and Application of a Simple “Easy To Operate” Propidium Monoazide-Crossing Priming Amplification on Detection of Viable and Viable But Non-culturable Cells of O157 Escherichia coli

Alterations in the Cell Wall of Rhodococcus biphenylivorans Under Norfloxacin Stress

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