Q-PCR Based Culture-Independent Enumeration and Detection of Enterobacter: An Emerging Environmental Human Pathogen in Riverine Systems and Potable Water

Overview

Journal Front Microbiol

Specialty Microbiology

Date 2016 Mar 1

PMID 26925044

Citations 8

Authors

Chandra B Patel

Rishi Shanker

Vijai K Gupta

Ram S Upadhyay

Affiliations

Soon will be listed here.

Abstract

The availability of safe and pristine water is a global challenge when large numbers of natural and anthropogenic water resources are being depleted with faster rate. The remaining water resources are severely contaminated with various kinds of contaminants including microorganisms. Enterobacter is one of the fecal coliform bacteria of family Enterobacteriaceae. Enterobacter was earlier used as an indicator bacterium along with other fecal Coliforms namely Escherichia coli, Citrobacter, and Klebsiella, but it is now known to cause various diseases in human beings. In this study, we have collected 55 samples from potable water and riverine system and proved their presence using their conserved sequences of 16S rRNA and 23S rRNA genes with the help of SYBR green real-time PCR, which showed very high specificity for the detection of Enterobacter. The Enterobacter counts in potable water were found to 1290 ± 32.89 to 1460 ± 39.42 cfu/100 ml. The Enterobacter levels in surface water were 1.76 × 10(4) ± 492, 1.33 × 10(4) ± 334, 1.15 × 10(4) ± 308, 2.56 × 10(4) ± 802, 2.89 × 10(4) ± 962, 8.16 × 10(4) ± 3443 cfu/100 ml; the levels of Enterobacter contamination associated with hydrophytes were 4.80 × 10(4) ± 1804, 3.48 × 10(4) ± 856, 8.50 × 10(4) ± 2074, 8.09 × 10(4) ± 1724, 6.30 × 10(4) ± 1738, 3.68 × 10(4) ± 949 cfu/10 g and the Enterobacter counts in sediments of the river, were 2.36 × 10(4) ± 703, 1.98 × 10(4) ± 530, 9.92 × 10(4) ± 3839, 6.80 × 10(4) ± 2230, 8.76 × 10(4) ± 3066 and 2.34 × 10(4) ± 732 cfu/10 g at the sampling Site #1, Site #2, Site #3, Site #4, Site #5, and Site #6, respectively. The assay could be used for the regular monitoring of potable water and other water reservoirs to check waterborne outbreaks.

Citing Articles

Waterborne pathogens detection technologies: advances, challenges, and future perspectives.

Oon Y, Oon Y, Ayaz M, Deng M, Li L, Song K Front Microbiol. 2023; 14:1286923.

PMID: 38075917 PMC: 10708915. DOI: 10.3389/fmicb.2023.1286923.

Characterization of Bacterial Communities from the Surface and Adjacent Bottom Layers of Water in the Billings Reservoir.

Marcondes M, Nascimento A, Pessoa R, Victor J, Duarte A, Clissa P Life (Basel). 2022; 12(8).

PMID: 36013459 PMC: 9409723. DOI: 10.3390/life12081280.

Bacterial Removal Efficiency of and From Surface Water: Laboratory-Based Cross-Sectional Study.

Tagele Haligamo D, Ejeso A, Beyene E Environ Health Insights. 2022; 16:11786302221111842.

PMID: 35846165 PMC: 9280837. DOI: 10.1177/11786302221111842.

Molecular Diagnostic Tools Applied for Assessing Microbial Water Quality.

Paruch L Int J Environ Res Public Health. 2022; 19(9).

PMID: 35564522 PMC: 9105083. DOI: 10.3390/ijerph19095128.

Propionic Acid and Sodium Benzoate Affected Biogenic Amine Formation, Microbial Community, and Quality of Oat Silage.

Jia T, Yun Y, Yu Z Front Microbiol. 2021; 12:750920.

PMID: 34819922 PMC: 8606646. DOI: 10.3389/fmicb.2021.750920.

References

Fish J, Pettibone G . Influence of freshwater sediment on the survival of Escherichia coli and Salmonella sp. as measured by three methods of enumeration. Lett Appl Microbiol. 1995; 20(5):277-81. DOI: 10.1111/j.1472-765x.1995.tb00445.x. View

Whitman R, Shively D, Pawlik H, Nevers M, Byappanahalli M . Occurrence of Escherichia coli and enterococci in Cladophora (Chlorophyta) in nearshore water and beach sand of Lake Michigan. Appl Environ Microbiol. 2003; 69(8):4714-9. PMC: 169104. DOI: 10.1128/AEM.69.8.4714-4719.2003. View

Ram S, Singh R, Shanker R . In silico comparison of real-time PCR probes for detection of pathogens. In Silico Biol. 2008; 8(3-4):251-9. View

Ristuccia P, Cunha B . Enterobacter. Infect Control. 1985; 6(3):124-8. DOI: 10.1017/s0195941700062810. View

Shrivastava R, Upreti R, Jain S, Prasad K, Seth P, Chaturvedi U . Suboptimal chlorine treatment of drinking water leads to selection of multidrug-resistant Pseudomonas aeruginosa. Ecotoxicol Environ Saf. 2004; 58(2):277-83. DOI: 10.1016/S0147-6513(03)00107-6. View

Sletvold H, Johnsen P, Simonsen G, Aasnaes B, Sundsfjord A, Nielsen K . Comparative DNA analysis of two vanA plasmids from Enterococcus faecium strains isolated from poultry and a poultry farmer in Norway. Antimicrob Agents Chemother. 2006; 51(2):736-9. PMC: 1797720. DOI: 10.1128/AAC.00557-06. View

Scott T, Rose J, Jenkins T, Farrah S, Lukasik J . Microbial source tracking: current methodology and future directions. Appl Environ Microbiol. 2002; 68(12):5796-803. PMC: 134426. DOI: 10.1128/AEM.68.12.5796-5803.2002. View

Gurtler J, Beuchat L . Performance of media for recovering stressed cells of Enterobacter sakazakii as determined using spiral plating and ecometric techniques. Appl Environ Microbiol. 2005; 71(12):7661-9. PMC: 1317380. DOI: 10.1128/AEM.71.12.7661-7669.2005. View

Singh K, Malik A, Mohan D, Sinha S . Multivariate statistical techniques for the evaluation of spatial and temporal variations in water quality of Gomti River (India)--a case study. Water Res. 2004; 38(18):3980-92. DOI: 10.1016/j.watres.2004.06.011. View

10.

Mullane N, Iversen C, Healy B, Walsh C, Whyte P, Wall P . Enterobacter sakazakii an emerging bacterial pathogen with implications for infant health. Minerva Pediatr. 2007; 59(2):137-48. View

11.

Jyoti A, Ram S, Vajpayee P, Singh G, Dwivedi P, Jain S . Contamination of surface and potable water in South Asia by Salmonellae: culture-independent quantification with molecular beacon real-time PCR. Sci Total Environ. 2009; 408(6):1256-63. DOI: 10.1016/j.scitotenv.2009.11.056. View

12.

Ishii S, Yan T, Shively D, Byappanahalli M, Whitman R, Sadowsky M . Cladophora (Chlorophyta) spp. harbor human bacterial pathogens in nearshore water of Lake Michigan. Appl Environ Microbiol. 2006; 72(7):4545-53. PMC: 1489363. DOI: 10.1128/AEM.00131-06. View

13.

Sinton L, Hall C, Braithwaite R . Sunlight inactivation of Campylobacter jejuni and Salmonella enterica, compared with Escherichia coli, in seawater and river water. J Water Health. 2007; 5(3):357-65. DOI: 10.2166/wh.2007.031. View

14.

Musgrove M, Northcutt J, Jones D, Cox N, Harrison M . Enterobacteriaceae and related organisms isolated from shell eggs collected during commercial processing. Poult Sci. 2008; 87(6):1211-8. DOI: 10.3382/ps.2007-00496. View

15.

Sherr E, Sherr B . Significance of predation by protists in aquatic microbial food webs. Antonie Van Leeuwenhoek. 2002; 81(1-4):293-308. DOI: 10.1023/a:1020591307260. View

16.

Byappanahalli M, Shively D, Nevers M, Sadowsky M, Whitman R . Growth and survival of Escherichia coli and enterococci populations in the macro-alga Cladophora (Chlorophyta). FEMS Microbiol Ecol. 2009; 46(2):203-11. DOI: 10.1016/S0168-6496(03)00214-9. View

17.

Hill D, Owens W, Tchounwou P . The impact of rainfall on fecal coliform bacteria in Bayou Dorcheat (North Louisiana). Int J Environ Res Public Health. 2006; 3(1):114-7. PMC: 3785686. DOI: 10.3390/ijerph2006030013. View

18.

Beuchat L . Ecological factors influencing survival and growth of human pathogens on raw fruits and vegetables. Microbes Infect. 2002; 4(4):413-23. DOI: 10.1016/s1286-4579(02)01555-1. View

19.

Kohn T, Nelson K . Sunlight-mediated inactivation of MS2 coliphage via exogenous singlet oxygen produced by sensitizers in natural waters. Environ Sci Technol. 2007; 41(1):192-7. DOI: 10.1021/es061716i. View

20.

Begum Y, Talukder K, Nair G, Qadri F, Sack R, Svennerholm A . Enterotoxigenic Escherichia coli isolated from surface water in urban and rural areas of Bangladesh. J Clin Microbiol. 2005; 43(7):3582-3. PMC: 1169094. DOI: 10.1128/JCM.43.7.3582-3583.2005. View