Long-Term Monitoring of Noxious Bacteria for Construction of Assurance Management System of Water Resources in Natural Status of the Republic of Korea
1Department of Biotechnology, College of Biomedical and Health Science, Konkuk University, Chungju 27478, Republic of Korea
2Department of Life Science, Graduate School, Kyonggi University, Suwon 16227, Republic of Korea
3DK EcoV Environmental Microbiology Lab., Cheonan 31075, Republic of Korea
4Environmental Infrastructure Research Department, Water Supply and Sewerage Research Division, National Institute of Environmental Research, Incheon 22689, Republic of Korea
5Department of Public Health Science, Graduate School of Public Health, Seoul National University, Seoul 08826, Republic of Korea
6Department of Parasitology and Tropical Medicine, School of Medicine, Inha University, Incheon 22212, Republic of Korea
J. Microbiol. Biotechnol. 2020; 30(10): 1516-1524
Published October 28, 2020
Copyright © The Korean Society for Microbiology and Biotechnology.
Life on Earth are greatly affected by the dynamics of climate system, especially the Earth’s surface climate. In particular, infectious pathogens are emerging as a source of issue as many aspects of public health accompanying the climate change are widely recognized [1, 2]. The term pathogen covers a wide range of disease agents, such as virus, bacteria, parasitic germs, and fungi that can affect human beings either directly or indirectly through influencing the habitat, environment, or by competing with other pathogens. Climate change is a global phenomenon and is expected to accelerate in the future, especially in situations where the extent of climate change on Korean peninsula is relatively large (
Campylobacterota, formerly identified as Epsilon proteobacteria, are a whole bunch of gram-negative motile bacteria found in manifold ecological habitats .
Climate change is causing water scarcity not only through increased temperatures and prolonged drought times but also through the degradation of water resources caused by increasing levels of pathogens and other contaminants posing significant health risks . Thus, there is clearly a strong need for establishing management strategies and constant monitoring the water resources based on the results of testing water for contamination from relevant sources. Herein, we report the results on long-term (six-year) surveillance of noxious bacteria (
Material and Methods
A catchment scale investigation of the prevalence of
Testing of Indicator Bacteria
Water samples were collected from each location.
Quality of the Collected Water Samples
Physiochemical parameters such as
Sample Collection and Analysis
The targeted bacteria were
Real-time PCR analysis was conducted with 10 ul of SYBR green master mix (Thermo Fisher Scientific) and 10 pM specific primer sets in a reaction volume of 20 ul using CFX96 Real-time PCR system (Bio-Rad Laboratories, USA). The primer set sequences and reaction conditions for each targeted noxious bacterium and the amplified target sizes represent in Table 2. The specificity of the primers was confirmed using a BLAST search in GenBank database from NCBI. For each bacterium tested in this monitoring, the BLASTn searches yield no solid match to any of the other identified bacterium reference sequences. Specificity tests were performed using conventional PCR techniques for each species or subspecies primer set against DNA samples from various bacterium strains . Matches between the cyclic quantification (Cq) value and each of noxious bacteria detection was verified, and positive estimation was determined for a single peak using the Cq value. The positive samples were analyzed and confirmed by sequencing the 16s rDNA fragments by Macrogen Inc. (Korea). Analysis of the derived nucleotide sequences was performed for matching genotypes using the NCBI-BLAST service to target the noxious bacteria.
Physicochemical Parameters in the Water Samples
There were slight variations in physiochemical parameters among the water sample collecting sites. The water temperature at the Guui region and Lake Juam tended to increase slightly, while no definite trend was observed for precipitation (Data not shown). The other collection sites did not show a definite tendency in the parameters of precipitation or water temperature. Table 3 summarizes the physiochemical parameters of water samples from the five sample collecting sites during the periods from August 2013 to February 2019. All the parameters other than precipitation and water temperature fluctuated continuously through the year. The
Indicator Bacteria and Water-Quality Monitoring Stations
The monitoring points in this study were selected for the water quality measurements due to the needs for long-term monitoring and management of Korean rivers and lakes and the links between the nearby water quality measuring network points according to the prediction scenario for climate change: Lake Soyang in the exceptional subtropical zone and the Guui region on the Han River in Seoul, Hyundo region (near Hyundo Bridge of Geum River at Shintanjin-dong in Daejeon), Lake Juam, and the Moolgeum region on Nakdong River in the subtropical zone.
In practice, it is impossible to enumerate all pathogens in water-source because of the absence of specific detection techniques. Thus, indicator bacteria including TC, FC and
Relationship between Physicochemical Parameters and Indicator Bacteria
Physicochemical parameters are among the major factors involved in the management and mitigation of non-point source pollution, and the effect of fecal contamination on the quality of water is a matter of quite concern. Information gained through the regular monitoring of water quality allows estimation of the likelihood pathogens-related waterborne disease . In general, indicator bacteria (TC, FC, and
Detection of Noxious Bacteria in Monitoring Sites
Most of the targeted bacteria were found in 77% of the samples and at least one of the target bacteria was detected (65%) (Fig. 1, Table 5). Among all the detected bacteria,
Figure 1. Total positive incidence cases of noxious bacteria surveillance in this study during the monitoring period.
Figure 2. Positive incidence cases of each noxious bacterium at each collecting site.
Statistical Analysis of Correlation between Indicator Bacteria and the Tested Noxious Bacteria
We performed correlation tests between the monitored six noxious bacteria and the tested three indicator bacteria (TC, FC,
The worldwide burden of infectious waterborne disease is considerable, and the bacterial pathogens are strongly resistant in the water environment and to most disinfectants. Some bacterial agents such as
Some of the noxious bacteria exhibited spatial and seasonal patterns at the collecting sites in this study. The presence of
According to the results of the monitoring in this study, the occurrences of noxious bacteria in water samples were dramatically decreased after April 2016. Although it is difficult to elucidate the specific cause, this could be attributed to the authorities’ effort, such as sewerage system management and social good-informed cognition, to improve the water quality. Korea achieved 92.1% penetration rate of sewage into the advanced countries through the first National Sewage Comprehensive Plan (NSCP) (2007-2015) through continuous expansion of sewage treatment facilities and sewage systems, improved sewage maintenance, enhancement of sewerage and sewerage management, establishment of water resource circulation utilization systems, and improved sewage treatment technology and sewage sludge treatment .
This study has a critical limitation. First of all, in some years during the study, the collection of surface water samples has limitations that have not been carried out as originally planned and thus we were not able to proceed with consistent sample collection and monitoring during the summer season. In addition, since this study was only based on the genetic analysis using PCR methods, we were not able to determine the infectivity and pathogenesis despite the positive detection. Nevertheless, this study was designed and practiced at these specific sites as a project of the National Institute of Environmental Research funded by the Ministry of Environment of the Republic of Korea. In fact, despite the growing interest in monitoring noxious microorganisms, it is difficult to find a case of research on their distribution and monitoring related to climate change at Korea or abroad. The Ministry of Environment of the Republic of Korea recognized the need for this research to provide public health security and secure drinking water stability because of water temperature rise, flooding, drought and heat waves due to climate change increase the prevalence of noxious microbes. The Ministry of Environment had set a Priority Management List (PML) of 20 noxious microbes in groups including TC bacteria, FC bacteria, pathogenic
In conclusion, it was not possible to determine the infectivity and pathogenicity on the six noxious bacteria examined in this study, and it was difficult to precisely identify any noticeable seasonal or regional effects. However, the fact that they were detected in the five Korea’s representative water environments comprising lakes, rivers, and drinking water collecting sites make it necessary to establish the chemical and biological analysis for noxious bacteria and sophisticated management systems in response to climate change. Thus, relying on predictive models and monitoring for timely warning can protect the health of the public.
This work was supported by a grant from the National Institute of Environmental Research (NIER-SP2018-309, TSK, OJR, and SSL) funded by the Ministry of Environment (MOE) of the Republic of Korea.
Conflict of Interests
The authors have no financial conflicts of interest to declare.
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