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Evaluation of Water and Sediment Quality in

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Article in Journal of Water and Environment Technology · June 2015

DOI: 10.2965/jwet.2015.207

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 Journal of Water and Environment Technology, Vol.13, No.3, 2015

Evaluation of Water and Sediment Quality in Open

Channels that Receive Effluent from Johkasou Facilities

Joni A. FAJRI1), Toshiro YAMADA2), Ahmad S. SETIYAWAN3), Fusheng LI4)

1) Graduate School of Engineering, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan
2) Faculty of Engineering, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan
3) Department of Environmental Engineering, Bandung Institute of Technology, Ganesha 10,
Bandung 40132, Indonesia
4) River Basin Research Center, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan

ABSTRACT
Treated water disposed from an alternative wastewater treatment system, named johkasou,
contains several contaminants that may affect the quality of the local water environment. Thus,
the evaluation of water quality, along with sediment content, may provide quantitative
information about the effects of johkasou effluent in the open channels. Here, the
physicochemical and microbial parameters were comprehensively examined in both the water
and sediments of a johkasou drainage channel and open channels. Throughout the 3-year study
period, the concentrations of physicochemical parameters were lower in the open channels
compared to the johkasou drainage channel because of the high mixing ratio of the flow rate
between the two channels. However, microbial parameters were consistently high in both the
water and sediments of open channels. Microbial concentrations were not significantly different
between upstream and downstream channels in any season, except for Escherichia coli (E. coli)
in winter (the concentrations of which increased in the downstream channel that received
johkasou effluent). Heterotrophic plate count bacteria and E. coli were positively correlated
between water and sediments, indicating that microbial indicators may interact with the water
and sediments in open channels.

Keywords: johkasou, microbial indicators, open channel

INTRODUCTION
Johkasou has been applied as an alternative wastewater treatment system in the rural
areas of Japan, in addition from other areas that do not have wastewater treatment plants
(Yang et al., 2010). Johkasou differs to septic tank and wetland systems because it
consists of a primary anaerobic and aerobic biological treatment unit, a sedimentation
tank, and a disinfection chamber, in which chlorine tablets are used (Ichinari et al.,
2008). The guidelines state that good quality of johkasou effluent must have less than
20 mg/L biochemical oxygen demand (BOD) (Nakajima et al., 1999). This type of
facility is considered an effective way of treating household wastewater before it is
discharged into a drainage or stream channel in residential areas.

The treated water of onsite domestic systems, including johkasou, may contain several
pollutants, such as organic substances, chemicals, nutrients, and microorganisms
(Savichtcheva et al., 2007; Wihters et al., 2011). However, many studies have
documented unsatisfactory quality of treated water from johkasou systems. For instance,
tandoku-shori johkasou has been reported as a major pollution source (e.g., organic
matter, and nutrients) of natural stream water in residential areas by discharging grey
water (Gaulke, 2006). In addition, insufficient removal of fecal indicators and
pathogenic bacteria in gappei-shori johkasou has been recorded at low temperatures

Address correspondence to Toshiro Yamada, Faculty of Engineering, Gifu University

Email: ymd@gifu-u.ac.jp
Received August 1, 2014, Accepted November 9, 2014.
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(i.e., during winter) (Kaneko et al., 2001). Furthermore, Setiyawan et al. (2014)
detected high levels of fecal indicators in the drainage channels of various types of
johkasou systems. Therefore, water quality monitoring in residential areas plays an
important role in evaluating the influence of johkasou effluent entering the local water
environment.

Analysis of water quality alone may underestimate the risk of human exposure to
potential pathogenic microorganisms in sediments. This is because sediments constitute
an important reservoir of microbial indicators in the water environment, including
stream channels in decentralized areas. A previous study documented a high density of
bacteria in the sediments of channels containing johkasou effluent (Helard et al., 2012).
Sediment may contain 100 – 1,000 times more fecal coliforms than the overlying water
(Bai and Lung, 2005). Compared to water, sediments contain organic substances and
optimal nutrient conditions for the development of microorganisms, particularly fecal
indicators, resulting in their long survival (Garzio-Hadzick et al., 2010).

Despite the importance of onsite domestic wastewater treatment systems in the water
cycle of local communities to protect public health, the physicochemical and microbial
quality of water and sediments in open channels that receive johkasou effluent are rarely
measured because most investigations to date have mainly focused on the treatment
performance of johkasou. Therefore, in the current study, we aimed to (i) evaluate how
johkasou effluent contributes to the quality of physicochemical and microbial
parameters in the water and sediments of open channels, and (ii) identify whether these
parameters are influenced by seasonal variation in the water flow of the open channels.

MATERIALS AND METHODS

Study site
The study site is located in Gifu, Japan, near a residential area that has a population of
around 250 inhabitants (Fig. 1). A total of 52 households use the johkasou facility in
this area (39 households [75%] use gappei-shori johkasou and 13 households [25%] use
tandoku-shori johkasou). Water samples were collected at six sampling points (SP)
along 1-m-wide open channels surrounding the residential area. The open channels
consisted of five sites and a small drainage channel, an outlet of 35 cm width, which
was the core sampling site (SP.4) that received effluent from 16 gappei-shori johkasou
facilities. The open channels were divided into upstream and downstream channels of
SP.4, of which three sampling sites were located upstream (SP.1, SP.2, and SP.3) and
two sampling sites were located downstream (SP.5 and SP.6). Sampling point 1 was
located in the outlet of another open channel, surrounded by a paddy field, which
received effluent from 10 tandoku-shori johkasou facilities and two gappei-shori
johkasou facilities from another residential area. Sampling point 2, which was located
25 m downstream of SP.1, was located in the channel that connected the SP.1 open
channel with another open channel that received ground water mixed with effluent from
23 gappei-shori johkasou facilities. Sampling point 3 was located 30 m after SP.2 and
received effluent from three tandoku-shori johkasou facilities. Sampling point 5 was
located in downstream of SP.4. Sampling point 6 was located 30 m after SP.5, which
was surrounded by a paddy field.

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Fig. 1 - Sampling site map in a residential area using johkasou facilities in Gifu
Prefecture, Japan. (Black dots represent the open channel sampling sites
(SP.1, 2, 3, 5, and 6) and a black square indicates the johkasou effluent
collected from the johkasou drainage channel (SP.4)).

Sample collection
Water samples were always collected in new 2 L polypropylene bottles that were rinsed
several times with the water sample of each site before sampling. The samples were
then placed in an ice box before being transported to the laboratory. All water samples
were placed in the refrigerator at 5°C and were immediately analyzed. The water
samples were collected on 14 occasions from November 2010 to January 2013.
Sampling was conducted on November 17 and December 20, 2010; March 15, August 8,
September 15, October 14, October 26, November 16, and December 15, 2011; March 8,
May 24, August 28, and November 23, 2012; and January 18, 2013.

Sediment samples were always collected in new 1 L polypropylene bottles that were
rinsed several times with the mixed liquor sediment of each site before sampling. The
samples were placed in an ice box before being transported to the laboratory. Sediment
samples were collected on 10 occasions from all sampling sites. Sampling was
conducted on November 17, 2010; March 15, September 15, October 26, November 16,
and December 15, 2011; March 8, May 24, and November 23, 2012; and January 18,
2013. Sediment samples were collected as sediment/water mixed liquor using a tube
with an inner diameter of 30 cm for SP.1, 2, 3, 5, and 6, and a small tube with an inner
diameter of 10 cm for SP.4. The mixed liquor from each site was then collected by
placing the sampling tube on the sediment bed, mixing the sediment with the overlying
water, and collecting the sediment mixed with the overlying water in the tube.

Analytical method
The following parameters were analyzed in the water samples: flow rate, water
temperature (WT), dissolved oxygen (DO), dissolved organic carbon (DOC), suspended

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solids (SS), biochemical oxygen demand (BOD), chemical oxygen demand (COD), total
nitrogen (TN), total phosphorus (TP), total chlorine, heterotrophic plate count bacteria
(HPC), total coliform (TC), Escherichia coli (E. coli), and DNA-based total bacteria
(DNA-based TB). The following parameters were analyzed in the sediment samples:
organic content, HPC, TC, E. coli, and DNA-based TB. In-situ measurements were
conducted for flow rate, WT, DO, and total chlorine.

Physicochemical analysis. The flow rate was calculated by multiplying the flow
velocity measured using an electromagnetic velocity meter (AEM1-D, JFE Advantech
Co. Ltd, Hyogo, Japan) in addition to water depth and channel width. The WT and DO
were measured directly at each site using the corresponding portable meters
(DKK-TOA, Tokyo, Japan). Total chlorine was also measured using the corresponding
pocket colorimeter (HACH, Dusseldorf, Germany). Measurements of SS, DOC, BOD,
COD, TN, TP, and total chlorine were conducted according to the standard method
(APHA-AWWA-WEF, 2005).

Microbiological analysis. Direct measurement of water and sediment samples was

conducted based on the standard method (APHA-AWWA-WEF, 2005) for each
microbial indicator related to HPC, TC, and E. coli. Heterotrophic plate count bacteria
were analyzed based on the plate count method using tryptone glucose yeast extract
(TGYE) agar and incubated at 20°C for 7 days. Fecal indicator bacteria-related TC and
E. coli were enumerated based on multiple tube fermentation method using ES
Colicatch 1000 (Eiken Chemical, Tokyo, Japan) and incubated at 37°C for 24 h.

Total number of bacteria was quantitatively measured based on the measurement of

DNA-based TB using Thermal Cycler Dice™ Real Time System TP800 (TaKaRa Bio
Inc., Shiga, Japan). The determination involved the following processes: DNA-based
TB extraction using Power Soil™ DNA Isolation Kit (MO BIO Lab. Inc., Carlsbad, CA,
USA), amplification with the universal 16S rDNA primer set (SYBR® Premix Ex Taq™,
Takara Bio Inc., Shiga, Japan), and detection by measuring the increase in fluorescence
caused by binding SYBR Green dye to double stranded DNA-based TB in a real-time
PCR (Zhou et al., 2007; Helard et al., 2012). The specificity of the assay was assessed
by the analysis of the melting curve (Fey et al., 2004). Melting was performed from 60
to 95°C at increments of 0.2°C/s. The concentration of DNA-based TB was then
converted to the bacterial number concentration by using E. coli as the surrogate. The
calibration curve for total bacteria in real-time PCR was obtained from extracted DNA
of E. coli according to the extraction method of E. coli 12F+ (No. 13965, NITE
Biological Resource Center, Japan) (Zhou et al., 2007). The DNA of E. coli was
quantified with a NanoVue UV/Visible Spectrophotometer (GE Healthcare,
Buckinghamshire, UK), and was then used to generate a standard curve using a 10-fold
dilution series in the range of 101 to 10–6 ng/µL. The range of melting temperature (Tm)
for the standard DNA was 86.5 – 87.5°C.

Statistical analysis
Significant differences in the physicochemical and microbial parameters of sampling
sites and seasons were calculated using one-way analysis of variance with Tukey’s
post-hoc test. The log-transformed was used as preparation data for all microbial
concentrations to avoid incorrect statistical analysis. The seasons were tabulated and

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categorized from all data, and were defined using a solar calendar for Gifu City area
(i.e., spring begins in mid-March, summer begins in mid-June, autumn begins in
mid-September, and winter begins in mid-December). The Spearman rank correlation
test was used to evaluate the correlations between microbial contamination in the water
and sediment. The statistical analyses were performed using IBM® SPSS® Statistic
version 21 and Excel 2010 with significance at the 95% confidence level.

RESULTS
Spatial variation in physicochemical and microbial parameters
The measurement results of the physicochemical parameters at six sampling sites are
shown in Table 1. The flow rate in the johkasou drainage channel (SP.4) was lower
(0.41 L/s) than that in the open channels (range: 11.7 – 22.6 L/s). Johkasou effluent
contained relatively higher levels of DOC, BOD, COD, TN, and TP than the open
channels (Tukey’s test for each parameter, P < 0.001), whereas DO was detected at a
low level. The concentrations of the physicochemical parameters were similar among
open channel sampling sites (P > 0.05).

The concentrations of HPC, TC, and E. coli significantly varied in both the water and
sediment of the johkasou drainage channel (Tukey’s test P < 0.01 and P < 0.05,
respectively). The geometric mean concentrations of microbial indicators were up to
two orders of magnitude higher in the johkasou drainage channel than in the open
channels (Table 2). However, the microbial concentrations were not significantly
different among the open channel sampling sites for both the water and sediment
samples (Tukey’s test P > 0.05), with concentrations being relatively higher in the
sediment samples (Table 3). The geometric mean concentrations of HPC, TC, E. coli,
and DNA-based TB in the water and sediments were similar among the open channel
sampling sites. The sediments at SP.4 had higher organic content than any of the open
channel sampling sites.

Table 1 - Results of physicochemical analyses in the water at six sampling points

during November 2010 – January 2013.
Upstream Johkasou* Downstream
Parameters
SP.1 SP.2 SP.3 SP.4 SP.5 SP.6
Water
Flowrate [L/s] 11.7 ± 14.4 (14) 13.7 ± 15.7 (11) 14.5 ± 15.7 (13) 0.41 ± 0.5 (14) 19.2 ± 20.5 (13) 22.6 ± 23.8 (14)
WT [oC] 16.3 ± 6.5 (14) 15.9 ± 6.1 (11) 16.3 ± 6.2 (13) 17.7 ± 6.9 (14) 16.5 ± 6.1 (13) 16.4 ± 6.2 (14)
DO [mg/L] 6.5 ± 0.5 (14) 6.5 ± 0.5 (11) 6.5 ± 0.5 (13) 4.4 ± 1.1 (14) 6.6 ± 0.5 (13) 6.5 ± 0.6 (14)
SS [mg/L] 8.0 ± 6.9 (14) 5.3 ± 6.9 (11) 6.2 ± 6.8 (13) 6.4 ± 7.6 (14) 9.3 ± 15.5 (13) 4.9 ± 4.0 (13)
DOC [mg/L] 3.8 ± 2.6 (14) 2.7 ± 2.3 (10) 2.4 ± 2.2 (13) 6.4 ± 6.8 (14) 2.1 ± 1.3 (12) 2.1 ± 1.1 (13)
BOD [mg/L] 2.6 ± 1.9 (7) 1.1 ± 0.4 (5) 1.4 ± 0.5 (7) 8.8 ± 2.9 (7) 1.4 ± 0.3 (7) 2.2 ± 2.1 (7)
COD [mg/L] 3.7 ± 3.3 (7) 1.4 ± 0.4 (5) 1.8 ± 0.7 (7) 9.1 ± 2.0 (7) 2.0 ± 0.8 (7) 2.6 ± 2.1 (7)
TN [mg/L] 3.1 ± 2.6 (12) 2.0 ± 2.1 (10) 1.4 ± 0.7 (12) 12.2 ± 7.2 (12) 1.7 ± 0.8 (12) 2.1 ± 1.3 (12)
TP [mg/L] 0.26 ± 0.26 (6) 0.16 ± 0.05 (4) 0.27 ± 0.23 (7) 1.68 ± 0.48 (7) 0.30 ± 0.29 (7) 0.25 ± 0.18 (6)
Total chlorine [mg/L] 0.05 ± 0.06 (9) 0.04 ± 0.03 (9) 0.03 ± 0.02 (9) 0.06 ± 0.04 (9) 0.06 ± 0.05 (9) 0.08 ± 0.1 (9)
The data is shown as "mean +/- standard deviation (number of samples)".
Asterisk mark (*) indicates the johkasou effluent collected in the johkasou drainage channel.

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Table 2 - Microbial concentrations in the water of six sampling points in the period of
November 2010 – January 2013.
HPC TC E. coli DNA-based TB
Sampling sites
CFU/mL MPN/100 mL MPN/100 mL cell/mL
Upstream
SP.1 8.0 × 104, 2.9 (14) 2.1 × 105, 2.9 (14) 3.3 × 103, 7.8 (14) 2.5 × 107, 10.6 (14)
SP.2 8.1 × 104, 3.4 (11) 2.3 × 105, 3.1 (11) 3.3 × 103, 5.1 (11) 1.1 × 107, 14.5 (11)
SP.3 1.0 × 105, 3.4 (13) 4.6 × 105, 3.6 (13) 1.9 × 103, 6.8 (13) 1.8 × 107, 13.7 (14)
Johkasou drainage
SP.4 4.8 × 105, 5.2 (14) 1.7 × 106, 3.9 (14) 6.7 × 105, 6.1 (14) 0.9 × 108, 14.0 (14)
Downstream
SP.5 1.2 × 105, 2.9 (12) 2.4 × 105, 4.1 (12) 5.0 × 103, 6.1 (12) 1.1 × 107, 9.8 (14)
SP.6 1.1 × 105, 3.5 (13) 2.2 × 105, 3.5 (13) 4.7 × 103, 9.3 (12) 2.3 × 107, 11.6 (13)
The data is shown as "geometric mean, geometric standard deviation (number of samples)".

Table 3 - Organic and microbial contents in the sediments at six sampling points in
the period of November 2010 – January 2013.
Organic content HPC TC E. coli DNA-based TB
Sampling sites
% CFU/g-dry weight MPN/g-dry weight MPN/g-dry weight cell/g-dry weight
Upstream
SP.1 10.2, 3.7 (10) 4.0 × 107, 4.8 (10) 1.3 × 106, 5.1 (9) 5.9 × 103, 2.4 (9) 2.3 × 108, 4.5 (9)
SP.2 15.3, 11.2 (9) 7
8.4 × 10 , 5.5 (9) 6
1.3 × 10 , 5.9 (8) 3
9.9 × 10 , 3.0 (8) 1.7 × 108, 4.4 (7)
SP.3 11.4, 4.7 (10) 7
9.4 × 10 , 3.0 (10) 6
1.3 × 10 , 3.5 (8) 3
9.5 × 10 , 2.2 (9) 1.2 × 108, 4.0 (8)
Johkasou drainage
SP.4 36.8, 16.5 (10) 2.2 × 109, 4.0 (10) 1.7 × 107, 58.3 (9) 3.0 × 105, 45.9 (9) 0.8 × 109, 12.0 (9)
Downstream
SP.5 19.8, 10.7 (10) 3.9 × 108, 5.7 (10) 1.2 × 107, 12.7 (9) 4.8 × 104, 9.6 (9) 8.3 × 107, 5.5 (8)
SP.6 14.2, 11.6 (10) 7
9.5 × 10 , 4.3 (10) 7
2.5 × 10 , 11.5 (9) 4
0.8 × 10 , 22.7 (9) 2.6 × 108, 3.6 (9)
The data is shown as "geometric mean (arithmetic mean for organic content), geometric standard deviation (standard deviation for organic
content) (number of samples)".

DISCUSSION
Spatial variation in physicochemical and microbial parameters
High BOD, COD, TN, and TP levels were recorded in the johkasou drainage channel,
because this site only received johkasou effluent from 16 houses using gappei-shori
johkasou. Based on the johkasou standard, the concentrations of BOD and TN at this
site were below the guideline limit (with less than 20 mg/L being recorded) (JECES,
2009). These results were consistent with a previous report stating that johkasou
effluent contained low concentrations of organic and nutrient pollution (Fujimura and
Nakajima, 1998).

Microbial parameters (HPC, TC, E. coli, and DNA-based TB) in the johkasou effluent
were consistently detected at high levels through study period, even though the
maximum total chlorine concentration was 0.11 mg/L in the accumulation johkasou
effluent (SP. 4) that flows through the drainage channel of 20 – 120 m in length. This
result indicates that total chlorine had insufficient contact time with johkasou effluent,
resulting in high microbial concentrations remaining in the johkasou drainage channel,
supporting a previous report (Nambu et al., 1996). In addition, TC concentration at the
johkasou drainage channel exceeded the guideline for johkasou effluent quality which
should be less than 3,000 CFU/mL (JECES, 2009). This result indicates that johkasou

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effluent may be a potential source of fecal contamination in local surface waters. High
microbial content was also detected in the sediment of the johkasou drainage channel,
which might be caused by the settling and growth of microbes due to the presence of
organic matter and nutrients in the sediments, combined with appropriate temperature
(Pote et al., 2009; Garzio-Hadzick et al., 2010).

Physicochemical concentrations at SP.1 were higher than the other open channel
sampling sites, whereas high concentrations of microbial parameters were detected
along the open channels. The high concentrations of the physicochemical and microbial
parameters at SP.1 may be caused by the introduction of untreated grey water from
tandoku-shori johkasou. Previous studies reported tandoku-shori johkasou as a major
source of water pollution in the local water environment because of the disposal of grey
water (Gaulke, 2006; Tanaka et al., 2007). This is because grey water originates from
household wastewater (excluding toilet water), which contains high levels of organic
matter, in addition to chemical, nutrient, and microbial contamination (Eriksson et al.,
2002; Ottoson and Stenstrom, 2003).

The TC concentrations along the open channels exceeded the Environmental Quality
Standard in Japan (MoE, 2014) which should be less than 5,000 MPN/100-mL.
Escherichia coli concentrations were slightly raised in the downstream channel after
receiving johkasou effluent (Table 2). Moreover, E. coli concentrations in this area were
one order of magnitude higher than in the major rivers of Gifu (Sasajima et al., 2008).
These results suggest that johkasou effluent may contribute to the fecal contamination
of the downstream channel rather than influencing physicochemical concentrations.
Furthermore, microbial contents were detected slightly higher in the sediments of the
downstream channel than in those of the upstream channel. The high concentrations of
microbes in the water may elevate the microbial content in the sediments by
resuspension, as well as increasing the settlement of certain microbes on the sediments
with settable particles.

Temporal variation in physicochemical and microbial parameters

Seasonal variation in physicochemical and microbial parameters was evaluated to
determine the presence of any seasonal effects on the johkasou drainage channel (SP.4),
and the open channels. Seasonal variations in water temperature (WT), flow rate, BOD,
and TN are shown in Fig. 2. Seasonal variation was recorded for WT, with WT being
high in summer (26.1°C) and low in winter (10.0°C) throughout the study period. Flow
rate along the open channels was high during spring (47.42 L/s) and low during winter
(4.85 L/s).

Out of all the monitored channels, the johkasou drainage channel had the lowest flow
rate all year round (less than 1 L/s). Compared to the open channels, this channel
contained high concentrations of BOD (range: 7.5 – 13.0 mg/L) and TN (range: 7.1 –
18.0 mg/L) (Fig. 2). The concentrations of BOD and TN in the open channels were
significantly different in winter compared to the other seasons (Tukey’s test P = 0.001
and P < 0.001, respectively). Minimum BOD and TN concentrations were recorded in
summer (range: 1.15 – 1.25 mg/L and 0.6 – 0.79 mg/L, respectively), while maximum
concentrations were recorded in winter (range: 2.61 – 2.92 mg/L and 3.15 – 3.3 mg/L,
respectively). This result indicates that the variation in seasonal flow rate affected

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physicochemical concentrations. Specifically, the mixing ratios (dilution factor)

between the johkasou drainage channel and the downstream channel in spring, summer,
autumn, and winter were 1:73, 1:63, 1:36, and 1:29, respectively.

Seasonal variation in microbial parameters (HPC, TC, E. coli, and DNA-based TB) in
the johkasou drainage channel and open channels is displayed in Fig. 3. The levels of
HPC, TC, and E. coli were slightly higher in winter compared to the other seasons in
both the effluent and sediments of the johkasou drainage channel. Seasonal variation of
microbial parameters in the johkasou effluent was only significant for E. coli
concentrations, with Tukey’s post-hoc test showed a significant difference between
winter and summer (P = 0.07). Concentrations of E. coli in the johkasou effluent were
recorded up to two orders of magnitude higher in winter compared to other seasons (Fig.
3c). This result indicates that low temperature in winter may negatively affect the
removal of fecal indicators at onsite domestic systems, including the johkasou facility
(Reinoso et al., 2008; Papadopoulos et al., 2011). In addition, the high concentrations of
fecal indicators in the johkasou effluent may cause high microbial sediment content in
the johkasou drainage channel due to settlement and growth.

As shown in Fig. 3, the concentrations of HPC, TC, and E. coli in the downstream
channel increased in winter. A significant difference for HPC and E. coli concentrations
along the open channels was obtained among seasons, with Tukey’s post-hoc test
showing significant variation in winter for both microbes (P = 0.042 and P = 0.017,
respectively). These results indicate that johkasou effluent influences conditions in the
downstream channel, particularly for E. coli during winter when the low mixing ratio
occurred. In addition, lower temperature inhibits the inactivation rate of fecal indicators
in natural water, enhancing the survival of fecal indicators (Flint, 1987).

Fig. 2 - Seasonal variations of (a) flow rate, (b) WT, (c) BOD, and (d) TN for the
upstream channel, the johkasou drainage channel, and the downstream
channel. Data in the graphs are means and standard deviations pooled from
the study period while the n and asterisk marks indicate the number of
samples and actual values of measurement, respectively.

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The contents of HPC, TC, E. coli, and DNA-based TB in the sediments of open
channels are presented in Fig. 3. The minimum and maximum concentrations of these
microbes were recorded in spring and autumn, respectively. Seasonal variation in the
microbial content of the sediment was statistically significant for HPC and TC levels.
Tukey’s post-hoc analysis showed that HPC and TC levels were significantly different
in spring compared to other seasons (P = 0.002 and P = 0.014, respectively). The
transportation and resuspension of microbes associated with sediment particles might be
caused by high flow rate during spring (Fig. 2a), and may explain the low microbial
content in sediment during this season.

Fig. 3 - Seasonal variation of microbial parameters (HPC, TC, E. coli, and

DNA-based TB) in the water (a – d) and in the sediments (e – h) of the
upstream channel, the johkasou drainage, and the downstream channel.
Data in the graphs are geometric means and geometric standard deviations
pooled from the study period while the n and asterisk marks indicate the
number of samples and actual values of measurement, respectively.

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Relationship between microbial indicators in the water and sediments

The relationship among the log-transformed concentrations of microbial indicators in
both the water and sediments was examined using Spearman rank correlation (Table 4).
Heterotrophic plate count bacteria were positively correlated with TC and E. coli
concentrations in both the water (r = 0.31 and 0.28, respectively; p < 0.05), and
sediment (r = 0.55 and 0.56, respectively; p < 0.05). These results indicate that high
concentrations of heterotroph bacteria may also be present fecal coliform group in the
water environment. Positive correlations were also observed between TC and E. coli in
both the water (r = 0.47; p < 0.05) and sediment (r = 0.61; p < 0.05). However, there
was no correlation between HPC and DNA-based TB in either the water or sediment,
which indicates that bacteria other than HPC may contribute to the amount of
DNA-based TB.

Heterotrophic plate count bacteria in the sediments was positively correlated with HPC,
TC, and E. coli in the water (r = 0.33, 0.40, and 0.38, respectively; p < 0.05). Positive
correlations were also observed for HPC, E. coli, and DNA-based TB between the water
and sediments (r = 0.33, 0.27, and 0.43, respectively; p < 0.05). These positive
correlations indicate that bacteria may interact with both the sediment and overlying
water. The dynamic existence of fecal indicators in the sediment may be due to the
association of fecal indicators with the settable particles under normal flow condition
(Characklis et al., 2005). Thus, fecal indicators in the sediment may reflect those in the
overlying water due to the resuspension of fecal indicators from the sediments during
storm runoff (Droppo et al., 2009). Therefore, it is recommended that the fecal indicator
content of the sediments should be monitored to determine the microbial behavior
during stormwater runoff.

Table 4 - Spearman rank correlation coefficients between microorganisms in the

water and sediment.
Microorganims in water Microorganisms in sediment
HPC TC E.coli DNA-based TB HPC TC E.coli DNA-based TB
Water
HPC
TC 0.31 (77)
E.coli 0.28 (76) 0.47 (76)
DNA-based TB 0.22 (77) 0.34 (77) 0.09 (76)
Sediment
HPC 0.33 (54) 0.40 (54) 0.38 (54) 0.15 (54)
TC 0.21 (50) 0.15 (50) -0.05 (50) 0.16 (50) 0.55 (53)
E.coli 0.07 (50) 0.12 (50) 0.27 (50) 0.26 (50) 0.56 (52) 0.61 (53)
DNA-based TB 0.24 (48) -0.03 (50) 0.19 (48) 0.43 (48) 0.13 (50) -0.06 (44) -0.25 (44)
Data shown are coefficient correlations. Numbers of samples are shown in parentheses.
Highlighted values are significant correlation at p < 0.05

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CONCLUSIONS
This study comprehensively monitored the physicochemical and microbial parameters
in the water and sediments of open channels receiving johkasou effluent. The open
channels that received johkasou effluent contained low concentrations of
physicochemical parameters (BOD, COD, TN, and TP) through all years, with no
significant difference among sampling sites. This result was due to the high mixing ratio
of flow rate between the open channels and the johkasou drainage channel during the
study period, except winter. Thus, it is important to retain high water loading in the
downstream network, especially in winter, to decrease the physicochemical
contamination. However, higher concentrations of microbial parameters (HPC, TC, E.
coli, and DNA-based TB) were detected in the effluent and sediments of the johkasou
drainage channel, with microbial concentrations in the open channels being an order of
magnitude lower than those in the johkasou drainage channel. Significant differences of
microbial concentrations in seasons were not found between the upstream and
downstream channel, except for E. coli during winter wherein the levels increased in the
downstream channel after receiving johkasou effluent. Thus, an improvement in the
disinfection process is required to remove the microbial contamination and enhance
johkasou performance. Positive correlations of HPC and E. coli concentrations were
observed between those in the water and sediment, which indicates that these microbes
interact in these two phases. In conclusion, it is advised that the fecal indicators in the
sediments are monitored to determine microbial behavior and its transport during
periods of stormwater runoff.

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