e c o l o g i c a l e n g i n e e r i n g 3 2 ( 2 0 0 7 ) 110

available at www.sciencedirect.com

journal homepage: www.elsevier.com/locate/ecoleng

Improving multi-soil-layer (MSL) system

remediation of dairy efuent
R. Pattnaik a , R.S. Yost a, , G. Porter a , T. Masunaga b , T. Attanandana c
a
b
c

Department of Tropical Plant and Soil Sciences, University of Hawaii, 3190 Maile Way, 102 St. John, Hawaii 96822, USA
Faculty of Life and Environmental Science, Shimane University, Matsue 690-8504, Japan
Department of Soil Science, Faculty of Agriculture, Kasetsart University, Jatujak, Bangkok 10900, Thailand

a r t i c l e

i n f o

a b s t r a c t

Article history:

Dairy efuent disposal is a serious problem in the Hawaiian Islands. Dairies often estab-

Received 23 December 2006

lish multiple settling lagoons to accumulate and store efuent. Occasionally, the overow

Received in revised form 5 July 2007

of lagoons leads to the transfer of nutrients, such as nitrogen (N) and phosphorus (P), and

Accepted 16 August 2007

other contaminants, to hydrologically associated surface, subsurface, and coastal waters.

This study was conducted to assess the removal of inorganic N and phosphate in dairy
efuent using multi-soil-layer (MSL) systems. Four MSL systems were constructed with two

Keywords:

replications of two treatments, which were Perlite and the Leilehua soil. Both materials were

Dairy efuent

used separately for forming an aerobic layer in the MSL systems, whereas an anaerobic layer

Dairy wastewater treatment

was formed from a mixture of charcoal, sawdust, iron lings and Honouliuli soil. The results

Multi-soil-layer system

of this study revealed that the removal of inorganic N was similar for the Leilehua and Perlite

Soil dynamics

MSL system, which was 2293% and 2196%, respectively. Phosphate removal was higher

Tropical soils

in the Leilehua MSL system (6499%) compared to the Perlite MSL system (997%). Addi-

Inorganic nitrogen removal

tional aeration increased the removal of phosphate by the Leilehua MSL system. Sucrose

effectiveness

application with a constant rate of aeration increased the removal of inorganic N both

Phosphate removal effectiveness

in the Leilehua and Perlite MSL systems and increased phosphate removal in the Perlite
MSL system. The study demonstrated that MSL systems have the potential to remove high
percentages of inorganic N and phosphate in dairy efuent enabling reuse of the water.
2007 Elsevier B.V. All rights reserved.

1.

Introduction

The dairy industry generates wastewaters characterized by

high concentrations of nutrients, organic contents, and
pathogens (USDA-SCS, 1992). The organic and nutrient content of dairy wastewaters depends upon the size, lactation,
and diet of the cow. In addition, dairy wastewater composition
is signicantly inuenced by the wastewater management,
climate, operating conditions, and types of ushing. Table 1
shows the levels of major nutrients in dairy wastewater. The
dairy industry is one of the major sources of waste efu-

Corresponding author. Tel.: +1 808 956 7066; fax: +1 808 956 3894.
E-mail address: rsyost@hawaii.edu (R.S. Yost).
0925-8574/$ see front matter 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.ecoleng.2007.08.006

ents in Hawaii and in the Continental U.S. (USDA-SCS, 1992).

Dairy efuent disposal is a serious problem in Hawaii and
other Pacic Islands (Farrell-Poe, personal communication,
2007). The problem is due to the conned aquifers and limited availability of water in Pacic Island environments. The
current method used in Hawaii to dispose dairy efuent is
large settling lagoons. Dairies often establish multiple lagoons
to accumulate and store efuent. Occasionally, the lagoons
overow, leading to the transfer of nutrients, such as nitrogen (N) and phosphorus (P), and other contaminants, which
can pollute surface, subsurface, and coastal waters. Efuents

e c o l o g i c a l e n g i n e e r i n g 3 2 ( 2 0 0 7 ) 110

Table 1 Dairy wastewater characteristics (Wright, 1996)

Potential pollutant
source
Milking center waste
Silage leachate
Barnyard runoff
Dairy manure

Biochemical oxygen
demand (mg L1 )
40010,000
12,00090,000
100010,000
20,000

high in N and P concentration can cause eutrophication of

the receiving waters, degrading water quality (Smith et al.,
1999). The Environmental Protection Agency (EPA) and State
Department of Health (DOH) have rules and regulations for
the disposal of dairy efuent (Hawaii State Department of
Health Wastewater Branch, 1996). Proper management of dairy
efuent is currently a serious problem in Hawaii, which has
increased the operation costs and reduced protability of
many island dairies. The inability of many dairy operators
to properly manage the efuent has forced more than 50%
of them to close during the last 10 years (C.N. Lee, personal
communication, 2006).
With rising environmental concerns and tighter governmental regulations, managing animal wastes in an
environmentally responsible and economically feasible way
can be a challenge. It is becoming imperative that new ways
of waste treatment be found that reduce excessive nutrients from dairy efuent and yet are efcient and reliable.
Some of the methods of dairy waste treatment include land
application (Caro-Costas et al., 1972; Valencia-Gica et al.,
2004), vegetative lter strips (Ikenberry and Mankin, 2000),
constructed wetlands (Schaafsma et al., 2000), aerobic and
anaerobic processes (Manariotis and Grigoropoulos, 2003), and
bioremediation (Prochaska and Zouboulis, 2003) which have
performed well, but their widespread use is limited because
they are either costly, require regular maintenance, require
large areas of land, or the wastewater must be pre-treated. The
multi-soil-layer (MSL) system is a promising alternative with
potential for reducing contamination associated with dairy
efuent.
The MSL system is a technology that uses natural soil in
a unit to facilitate wastewater treatment (Wakatsuki et al.,
1993). This has been successfully developed in Japan and Thailand to treat domestic and restaurant wastewater as well as
polluted river water (Wakatsuki et al., 1993; Luanmanee et
al., 2001). The system reduces levels of inorganic contaminants such as nitrate, ammonium, and phosphate, as well

Nitrogen
(mg kg1 )

Phosphorus
(mg kg1 )

80900
4400
502500
5600

25170
500
5500
900

Volume gallons
(100 cows y1 )
73,000
105,000
80,000
660,000

as organic contaminants as measured by high COD (chemical oxygen demand) and BOD (biological oxygen demand).
This is a biphasic layered system that uses locally available
materials such as soil, iron particles, jute or sawdust, charcoal, and zeolite or alternative materials (Attanandana et al.,
2000; Luanmanee et al., 2001). Two layers that comprise MSL
systems are aerobic and anaerobic. Aerobic layers consist of
zeolite or Perlite alternated with anaerobic layers of soil mixture blocks. The efciency of the MSL system in purifying
wastewater depends on the relative effectiveness of aerobic
and anaerobic layers (Wakatsuki et al., 1993; Attanandana et
al., 2000). The aerobic layer enhances nitrication, oxidation
and precipitation of mobile ferrous iron to high-surface area
ferric oxide, enhancing phosphorus sorption (Wakatsuki et al.,
1993). In the anaerobic layer of the soil mixture block, nitrate
is transformed into nitrous oxide and nitrogen gas (denitried) and ferric iron is reduced to the more mobile ferrous
iron, which moves out of the anaerobic layer (Wakatsuki et
al., 1993). Although an appropriate amount and timing of aeration is necessary (Luanmanee et al., 2002), the maintenance
of an MSL system is simple and the effective life of such systems was estimated to be longer than 10 years (Luanmanee et
al., 2002).
Although various types of wastewater treatments have
been treated successfully using the MSL system in Japan and
Thailand, to-date, no MSL system has been tested or adapted
for the remediation of dairy efuent. In addition, there is
not much information available on the reliability, consistency,
and nutrient removal efciency of MSL systems. Thus, it is of
interest to determine whether the MSL system can remediate dairy efuent. This study was conducted to (a) investigate
the potential of the MSL systems in remediating dairy efuent, (b) compare the removal of inorganic N and phosphate,
between MSL systems with the aerobic layers made from
Leilehua soil or Perlite, and (c) evaluate the effect of aeration
and sucrose additions on inorganic N and phosphate removal
efciency.

Table 2 Selected physical properties of Leilehua and Honouliuli soils

Series

Clay (<0.002)
(% of <2 mm
mineral soil)

Silt (0.0020.05)
(% of <2 mm
mineral soil)

Sand (0.052)
(% of <2 mm
mineral soil)

Water holding
capacity (% of <2 mm
mineral soil)
33 kPa

Leilehua
Honouliuli

58.5
58.4

Source: Soil Survey Staff (2006).

33.6
34.8

7.9
6.8

n/a
30.2

Bulk density
(g cm3 )

Particle
density
(g cm3 )

1500 kPa
33.2
22.3

0.97
1.31

2.88
2.93

e c o l o g i c a l e n g i n e e r i n g 3 2 ( 2 0 0 7 ) 110

2.

Materials and methods

2.1.

Experimental site and design

essary (Fig. 1). An array of efuent emitters was installed on

the top (80 cm from the bottom) of the aerobic and anaerobic layers through which the dairy efuent was discharged
into the system (Fig. 1).

The experimental site was located in Waianae, latitude 21 27 ,

longitude 158 11 on the west shore of the island of Oahu,
Hawaii. Average maximum and minimum daily temperatures
of the area are 28 C (83 F) and 16 C (61 F) (Hobo Weather
Station, 20012003). The experiment was conducted using
dairy efuent from the third settling lagoon of an efuent
waste management system. Four MSL systems were constructed, comprising two treatments with two replications
each, arranged in a completely randomized design (CRD). Perlite or Leilehua soil (A horizon, Typic Kanhaplohumult) was
used for the aerobic layer in the two treatments. The anaerobic
layer for both treatments consisted of a mixture of charcoal,
sawdust, iron lings and the Honouliuli soil (A horizon, Typic
Chromustert). The physical and chemical properties of both
the Leilehua and Honouliuli soils used for the experiment are
given in Tables 2 and 3.

2.2.

2.2.1.

Application rates

The dairy efuent was directly pumped from the lagoon, ltered using a 0.0254 m plastic disc lter (140 mesh) to remove
the larger particles, and discharged into the MSL system.
Three application rates of efuent were applied to the system according to the performance of the system. An initial
ow rate of 80 L day1 (505 L m2 day1 ) was applied to each
of the system from 18 April to 3 November 2005. Efuent
was applied through drip irrigation emitters during approximately 20 h day1 . The ow rate was reduced to 40 L day1
(252 L m2 day1 ) on 3 November 2005 and continued until 20
April 2006. Then from 20 April to 10 July 2006 the ow rate was
again reduced to 28 L day1 (178 L m2 day1 ).

2.2.2.

Aeration rates

Different aeration rates were applied to the systems. Aeration was not applied to the systems until the 10th month of
the study. The systems were aerated at a rate of 28 L min1
from 10 February to 13 April 2006. The aeration was increased
to 31 L min1 from 14 to 27 April 2006. The aeration rate was
decreased to 17 L min1 for 1 week, from 28 April to 4 May 2006
followed by a rate of 11 L min1 until 18 May 2006. The aeration rate was increased again to 23 L min1 from 19 May until
10 July 2006.

MSL systems and operations

Cross-section composition of the overall MSL systems is presented in Fig. 1. Each of the MSL system consisted of a
high-density polyethylene (HDPE) corrugated sewage pipe
with 45.7 cm interior diameter by 1 m in height with a crosssectional area of approximately 0.1648 m2 (Fig. 1). A 25.4 mm
PVC pipe was installed at the base of each of the upright HDPE
pipe to discharge the MSL treated efuent from the system.
A layer of gravel (5 cm) was placed at the bottom of the
upright pipes to facilitate system discharge. Each system was
assembled from seven alternating layers of soil mixture blocks
(anaerobic layers) and eight layers of Leilehua soil or Perlite
(aerobic layers) (Fig. 1). Each of the soil mixture blocks consisted of Honouliuli soil mixed with nely ground charcoal,
ne sawdust, and approximately 1 mm diameter iron lings at
the ratio of 7:1:1:1 by dry weight. The soil mixture was evenly
mixed using an concrete mixer and packed into two sizes of
pre-stitched burlap bags, approximately 5 cm 10 cm 22 cm
and 5 cm 10 cm 38 cm. The particle sizes of both the Leilehua soil and the Perlite ller were less than or equal to 4 mm.
An aeration pipe was installed approximately 50 cm from the
bottom for the subsequent infusion of air whenever it was nec-

2.2.3.

Sucrose additions

An additional source of carbon in the form of a sucrose solution was applied to the MSL system beginning at the end
of the 12th month of the study in attempt to improve system performance. The sucrose solution was calculated based
on the amounts needed for a stoichiometric reduction of the
expected oxygen content of the MSL system.
The percent pore space was calculated rst from the bulk
densities and particle densities of Leilehua and Honouliuli soil,
and Perlite. Then the amount of air space was calculated from
the volume of each system. The amount of oxygen was calculated from the amount of air space and the amount of oxygen
in the air. The amount of sucrose was calculated based on
the stoichiometric reaction equation, which shows how much
sucrose is needed for the microorganisms to consume the spe-

Table 3 Chemical properties of Leilehua and Honouliuli soils

Soil
Leilehua
Honouliuli

pH
(H2 O, 1:1)
4.8
6.9

Source: Soil Survey Staff (2006).

a
b
c
d
e

Organic carbon.
Total nitrogen.
Dithionite-citrate extractable iron.
Ammonium oxalate iron.
Guo and Yost (1998).

OCa (% of
<2 mm)

TNb (% of
<2 mm)

Dithionitec
(% of <2 mm)

Oxalated (% of
<2 mm)

2.61
0.74

0.233
0.11

6.4
7.5

1.04
n/a

P sorbede
(mg kg1 )
1400
100

e c o l o g i c a l e n g i n e e r i n g 3 2 ( 2 0 0 7 ) 110

Fig. 1 Cross-sections of the MSL systems (Leilehua and Perlite).

cic amount of oxygen:

C12 H22 O11 + 12O2 12CO2 + 11H2 O

The calculated concentration of sucrose solution was 19 g

(0.055 moles per 500 mL) for the Leilehua system and 22 g
(0.064 moles 500 mL) for the Perlite system.

2.3.
Finally, the application of sucrose was made as a solution
mixed with the incoming efuent and applied based on the
efuent retention time of the MSL systems.

Analytical methods

Samples were taken every week except during the period

December 2005 to 14 January 2006. After ltering samples

e c o l o g i c a l e n g i n e e r i n g 3 2 ( 2 0 0 7 ) 110

Table 4 Dairy efuent used in this experiment, in comparison with data from other dairy lagoons in Hawaii
Source

pH

Dairy aa
Dairy bb
This experimentc
This experimentd
This experimente

8.2
8.1
8.4
7.77
8.90

EC
TSS
(mS cm1 ) (mg L1 )
6.4
3.0
3.6
3.22
3.62

n/a
1000
400
n/a
320

TN
(g mL1 )
395.7
119
44.6
20
5.39f

NO3 -N
(g mL1 )

NH4 + -N
(g mL1 )
183.8
108
43.16
17.05
5.26

n/a
5.76
1.30
2.86
0.12

TP
(g mL1 )

IP
(g mL1 )

14.5
16.8
21.39
17.76
NA

n/a
n/a
13.21
5.72
6.08

COD
(mg L1 )
n/a
n/a
482
447
710

EC: electrical conductivity; TSS: total suspended solid; TN: total nitrogen; TP: total phosphorus; IP: inorganic phosphate; COD: chemical oxygen
demand.
a
b
c
d
e
f

Analysis of lagoon efuents from various nutrient streams (Fukumoto et al., 2000).
Valencia-Gica et al. (2004).
One month before running the experiment, 3 March 2005.
The beginning of the experiment, 2 May 2005.
The end of the experiment, 10 July 2006.
Total inorganic N (summation of NH4 + -N and NO3 -N).

were analyzed for the following: ammonia nitrogen (NH4 + -N),

nitrate nitrogen (NO3 -N), and inorganic phosphate. Ammonia nitrogen was measured using the salicylate method
(Mulvaney, 1996a). Nitrate nitrogen was measured using the
cadmium reduction method (Mulvaney, 1996b). Total efuent
nitrogen consisted of 98% ammonium and about 1% nitrate.
The total inorganic nitrogen (Inorganic N) was approximated
as the summation of NH4 + -N and NO3 -N. The ascorbic acid
method was used to measure total inorganic phosphate (Kuo,
1996).

2.4.

(3) the combination of these two improvements.

3.1.

The analysis of the efuent was compared with other dairy

efuents in Hawaii (Fukumoto et al., 2000; Valencia-Gica et
al., 2004) (Table 4). The concentration of total N, NH4 + -N, and
NO3 -N was lower in the efuent use in this experiment than
the other dairy efuent. This might be a result of using efuent
from the third and last settling of the lagoon system, which
was more diluted than that from the rst lagoon.

Statistical analysis
3.2.

The percentage removal of inorganic N and phosphate

between the Leilehua and Perlite MSL systems were compared using Sigma Plot version 9 (Sigma Plot, 2004). Data for
selected intervals of time corresponding to specic treatments
were also analyzed using the Statistical Analysis Software, SAS
PROC MIXED Repeated Measures ANOVA and Least Square
means (LSmeans) (SAS, 2004) (Littell et al., 1996, 1998; SAS,
2004).

3.

Characteristics of dairy efuent

First phase (year 2005)

The effectiveness of the MSL systems in removing inorganic N

and phosphate was compared over a 6-month period (5 May5
October) (Figs. 2 and 3).
The MSL systems were not signicantly different in percentage removal of inorganic N (P > 0.1) (Table 5). However,
the percentage removal of inorganic N was signicantly differ-

Results

The percentage removal of inorganic N and phosphate are

discussed in two phases. The rst phase data occurred from
May to October 2005, when the system was operated at constant conditions and the second phase data from January to
July 2006, where specic treatments were applied. Although
samples were collected in the rst phase from October to
December 2005, the data were not included in the analysis
because of system malfunction. There was a 6-week pause (2
December 2005 to 14 January 2006) between the two phases
due to mechanical problems and also due to a suspected build
up of biolms. In the second phase aeration and sucrose additions were compared in an attempt to increase the efciency
of the MSL systems. Three possible improvements were tested
in the second phase:
(1) effect of increased aeration,
(2) effect of sucrose addition with a constant rate of aeration,
and

Fig. 2 Removal of inorganic N in the Leilehua and Perlite

MSL systems as affected by time. In this gure 2D
indicates a 2-day pause; 3D a 3-day pause; 6D a 6-day
pause; 8D a 8-day pause; 12D a 12-day pause.

e c o l o g i c a l e n g i n e e r i n g 3 2 ( 2 0 0 7 ) 110

Fig. 3 Removal of phosphate in the Leilehua and Perlite

MSL systems as affected by time. In this gure 2D
indicates a 2-day pause; 3D a 3-day pause; 6D a 6-day
pause; 8D a 8-day pause; 12D a 12-day pause.

Table 5 A comparison of the effect of time and MSL

system on inorganic N removal as analyzed by SAS Proc
Mixed repeated measures analysis
Factor
MSL systems
Time
MSL systems time

Probability of signicance (P)

0.8303
<0.0001
0.9474

ent over time for both the MSL systems (P < 0.0001) (Table 5).
The non-signicant interaction indicates that the MSL systems behaved similarly in percentage removal of inorganic
N (P > 0.1) (Table 5). The inorganic N removal by the Leilehua
MSL system and the Perlite MSL system ranged from 22 to 93%
(LSmean of 61.94) and 21 to 96% (LSmean of 63.40), respectively
(Fig. 2).
The percentage removal of phosphate was signicantly different by both the MSL systems (P < 0.05) (Table 6). The Leilehua
MSL system was more effective in removing phosphate than
the Perlite MSL system. There was also a signicant difference in percentage removal of phosphate over time (P < 0.001)
(Table 6). The signicant interaction indicates that there was
a decrease in percentage removal of phosphate in the Perlite
MSL system (P < 0.1) (Table 6). The percentage removal of phosphate by the Leilehua MSL system (6499%) (LSmean of 92.70)
was greater than the Perlite MSL system (997%) (LSmean of
59.41) (Fig. 3).

Fig. 4 Removal of inorganic N in the Leilehua and Perlite

MSL systems as affected by sucrose addition and different
rates of aeration.

3.3.

Second phase (year 2006)

3.3.1.

The effect of aeration

A comparison was made between no aeration and two different rates of aeration (28 L min1 and 31 L min1 ) in removal
of inorganic N and phosphate during a sampling period of 19
January to 27 April 2006 (Figs. 4 and 5).
The percentage removal of inorganic N was not signicantly different between the MSL systems (P > 0.1) (Table 7).
There was no signicant difference in percentage removal of
inorganic N with aeration for both the MSL systems (P > 0.1)
(Table 7). The non-signicant interaction indicates that the
two MSL systems behaved similarly in percentage removal of
inorganic N (P > 0.1) (Table 7). The removal of inorganic N by the
Leilehua system and the Perlite system ranged from 8 to 61%
(LSmean of 29.34) and 10 to 73% (LSmean of 33.10), respectively
(Fig. 4).

Table 6 A comparison of the effect of time and MSL

system on phosphate removal as analyzed by SAS Proc
Mixed repeated measures analysis
Factor
MSL systems
Time
MSL systems time

Probability of signicance (P)

0.0292
0.0004
0.0997

Fig. 5 Removal of phosphate in the Leilehua and Perlite

MSL systems as affected by sucrose additions and different
rates of aeration.

e c o l o g i c a l e n g i n e e r i n g 3 2 ( 2 0 0 7 ) 110

Table 7 A comparison of the effect of aeration and MSL

system on inorganic N removal as analyzed by SAS Proc
Mixed repeated measures analysis

Table 10 A comparison of the effect of sucrose and MSL

system on phosphate removal as analyzed by SAS Proc
Mixed repeated measures analysis

Factor

Factor

Probability of signicance (P)

MSL systems
Aeration
MSL systems aeration

0.8008
0.8214
0.4330

Probability of signicance (P)

MSL systems
Sucrose
MSL systems sucrose

0.0342
0.1325
0.0980

Table 8 A comparison of the effect of aeration and MSL

system on phosphate removal as analyzed by SAS Proc
Mixed repeated measures analysis

Table 11 A comparison of the effect of different rates of

aeration and MSL system on inorganic N removal as
analyzed by SAS Proc Mixed repeated measures analysis

Factor

Factor

Probability of signicance (P)

MSL systems
Aeration
MSL systems aeration

0.0088
0.0782
0.0141

The percentage removal of phosphate was signicantly

different between the MSL systems (P < 0.01) (Table 8). The
Leilehua MSL system was more effective in percentage
removal of phosphate than the Perlite MSL system. Changes
in aeration resulted in a signicant difference in percentage
removal of phosphate (P < 0.1) (Table 8). The signicant interaction indicates that the MSL systems behaved differently in
the removal of phosphate (P < 0.05) (Table 8). The removal of
phosphate by the Leilehua MSL system ranged from 42 to 91%
(LSmean of 66.64) and was greater than the Perlite MSL system
(1141%) (LSmean of 27.22) (Fig. 5).

3.3.2.

The combined effect of sucrose with aeration

A comparison was made between the non-sucrose and

sucrose applications with constant aeration in removal of inorganic N and phosphate from 16 February to 13 April and 25 May
to 10 July 2006 (Figs. 4 and 5).
The MSL systems were not signicantly different in percentage removal of inorganic N (P > 0.1) (Table 9). However,
there was a signicant increase in percentage removal of inorganic N with sucrose additions for both the MSL systems
(P < 0.1) (Table 9). The non-signicant interaction indicates that
the MSL systems behaved similarly in percentage removal of
inorganic N (P > 0.1) (Table 9). The inorganic N removal by the
Leilehua and the Perlite MSL system ranged from 9 to 89%
(LSmean of 48.77) and 10 to 92% (LSmean of 53.36), respectively
(Fig. 4).
The MSL systems differed signicantly in the percentage
removal of phosphate (<0.05) (Table 10). There was no overall signicant difference in percentage removal of phosphate
with the sucrose application (P > 0.1) (Table 10). However, the
signicant interaction indicates that the percentage removal

Probability of signicance (P)

MSL systems
Aeration
MSL systems aeration

0.8640
0.3546
0.3249

of phosphate was increased in the Perlite system (P < 0.1)

(Table 10). The removal of phosphate by the Leilehua MSL system ranged from 59 to 93% (LSmean of 76.31) and was more
effective than the Perlite MSL system (1175%) (LSmean of
46.13) (Fig. 5).

3.3.3.

The effect of different rates of aeration

Three different rates of aeration (11 L min1 , 17 L min1 , and

23 L min1 ) were compared in removal of inorganic N and
phosphate between a sampling period of 4 May to 10 July 2006
when sucrose was added (Figs. 4 and 5).
The MSL system performance was not signicantly different in the percentage removal of inorganic N (P > 0.1) (Table 11).
There was no signicant difference in percentage removal of
inorganic N with different rates of aeration between the MSL
systems (P > 0.1) (Table 11). The non-signicant interaction
indicates that the MSL systems behaved similarly in percentage removal of inorganic N (P > 0.1) (Table 11). The removal of
inorganic N by the Leilehua and the Perlite systems ranged
from 31 to 89% (LSmean of 60.33) and 43 to 92% (LSmean of
62.24), respectively (Fig. 4).
There was a signicant difference observed in percentage removal of phosphate between the MSL systems (P < 0.1)
(Table 12). There was no signicant difference in removal of
phosphate with different rates of aeration (P > 0.1) (Table 12).
The non-signicant interaction indicates that the MSL systems behaved similarly in removal of phosphate (P > 0.1)
(Table 12). The percentage removal of phosphate by the Leilehua MSL system ranged from 59 to 93% (LSmean of 73.71) was
greater than the Perlite MSL system (1775%) (LSmean of 46.77)
(Fig. 5).

Table 9 A comparison of the effect of sucrose and MSL

system on inorganic N removal as analyzed by SAS Proc
Mixed repeated measures analysis

Table 12 A comparison of the effect of different rates of

aeration and MSL system on phosphate removal as
analyzed by SAS Proc Mixed repeated measures analysis

Factor

Factor

MSL systems
Sucrose
MSL systems sucrose

Probability of signicance (P)

0.6583
0.0539
0.8913

MSL systems
Aeration
MSL systems aeration

Probability of signicance (P)

0.0937
0.1373
0.2538

e c o l o g i c a l e n g i n e e r i n g 3 2 ( 2 0 0 7 ) 110

Fig. 6 The relationship between P sorbed and soil solution

P in Leilehua and Honouliuli soil.

4.

Discussion

4.1.
Performance of the MSL system in removing
inorganic N
The efciency of the MSL systems in removing inorganic N
from dairy efuent was not signicantly different throughout the study (years 2005 and 2006). Both MSL systems used
for the aerobic layers, whether Leilehua or Perlite, were similarly effective in removing inorganic N. However, the MSL
systems were signicantly different in removal of inorganic
N over time in 2005. The removal rate of inorganic N in efuent decreased over time. We hypothesized the decrease might
be due to inadequate aeration in the aerobic layer or decreased
microorganism-available carbon in the anaerobic layer and
tested this by adding supplemental aeration and sucrose (as
a carbon source) in 2006. The systems were not signicantly
different in removal of inorganic N with supplemental aeration. However, the removal of inorganic N was signicantly
increased in both the MSL systems with the application of
sucrose. The increased removal rate was likely due to the
additional carbon provided by sucrose applications, which
enhanced microbial activity and thus increased the denitrication in both MSL systems. The removal of inorganic N was
not signicantly different with different rates of aeration in
2006. There were some pauses in efuent delivery by the MSL
systems in 2005 and sudden drops in removal of inorganic N
seemed to related to these pauses.

4.2.
Performance of the MSL system in removing
phosphate
The efciency of the MSL systems in removing phosphate from
dairy efuent varied signicantly during the study (years 2005
and 2006). The Leilehua MSL system was consistently more
effective in removing phosphate than the Perlite MSL system.
This was probably because of the high P sorption capacity of
the Leilehua soil in the Leilehua MSL system (1600 g P g1 soil)
(Fig. 6) which adsorbs phosphate from the efuent. The
removal of phosphate was signicantly decreased over time
by the Perlite MSL system in 2005. We hypothesized that the
decrease in removal of phosphate by the Perlite system might

be a result of decreased microorganism-available carbon in

the anaerobic layer related to reduced iron movement into the
aerobic layer. Supplemental aeration and carbon (as sucrose)
were applied in 2006 to increase the efciency of the MSL
systems in removing phosphate. The percentage removal of
phosphate was signicantly increased with the application of
sucrose in the Perlite MSL system. This might be because the
additional sucrose carbon increased the activity of microorganisms resulting in more oxygen consumption and enhanced
reducing conditions in the anaerobic layer chemically reducing and moving iron into the aerobic layer where it could
precipitate as ferric iron and sorb the phosphate in efuent.
The hypothesized decrease in microorganism-available carbon in 2005 seems to be supported by the sharp increase
in percentage removal of phosphate by the Perlite MSL system with sucrose applications observed in 2006. The removal
of phosphate was signicantly increased with supplemental
aeration by the Leilehua MSL system. This might be because
of the sufcient aeration in the Leilehua MSL system oxidized ferrous iron to ferric iron in the aerobic layer, leading
to higher adsorption of phosphate by the soil colloids. The
sucrose application did not increase the already high removal
of phosphate in the Leilehua MSL system. The removal of
phosphate was not signicantly different with different rates
of aeration when comparing the two MSL systems in 2006.
The systems consistently removed phosphate with different
rates of aeration. Thus, from the results of supplemental aeration and sucrose applications it appears that the phosphate
removal mechanism is likely different between the two MSL
systems. The removal of phosphate in the Leilehua MSL system was mainly due to sorption by iron in the aerobic layer,
whereas in the Perlite MSL system it appears to be due to three
steps, solubilization in the anaerobic layer, movement into the
aerobic layer, and precipitation as ferric oxide.

4.3.

Use of MSL-treated efuent

The Hawaii State Department of Health has three different

categories of recycled waterR-1, R-2, and R-3 waterwhich
are listed in Table 13 with specic criteria (Hawaii State
Department of Health, 2002). R-1 is the highest quality recycled water. It has been ltered and disinfected. It can be used
in any form of irrigation served by xed irrigation systems supplied by buried piping for turf and landscape irrigation of golf
courses, parks, elementary schools, roadsides, and residential
property where managed by an irrigation supervisor (Hawaii
State Department of Health, 2002). R-2 is a slightly lower
quality recycled water. It is secondary (biologically) treated
wastewater that has also been ltered and disinfected (Hawaii
State Department of Health, 2002). Its use requires more caution and restrictive controls than R-1 water. R-3 is the least
pure class of recycled water. R-3 quality water is wastewater
that has been treated to the secondary level. It can only be
used for irrigation at places where people rarely go (Hawaii
State Department of Health, 2002).
The average concentration of NO3 -N and phosphate, and
fecal coliform colonies in MSL-treated efuent of our study
is given in Table 14. If we compare our study with the recycled water requirements of State Department of Health in
Hawaii, the MSL-treated efuent comes in as R-3 water. MSL-

e c o l o g i c a l e n g i n e e r i n g 3 2 ( 2 0 0 7 ) 110

Table 13 Recycled Water Standards (Hawaii State Department of Health, 2002)

Type of recycled water

Treatment

Recycled water quality

Recycled water monitoring

R-1

Oxidizeda
Filteredb
Disinfectedc

23 fecal coliform/100 mL
Nitrate 10 mg L1
Total phosphorus 1.0 mg L1

Coliform: no more than one sample in any 30-day period

R-2

Oxidized
Filtered
Disinfected

200 fecal coliform/100 mL

Nitrate 10 mg L1
Total phosphorus 1.0 mg L1

Coliform: no more than one sample in any 30-day period

R-3

Oxidized
Secondary
Undisinfected

a
b
c

Wastewater in which the organic matter has been stabilized.

The passing of wastewater through natural undisturbed soils or lter media such as sand.
The destruction, inactivation, or removal of pathogenic microorganisms by chemical, physical, or biological means. Disinfection may be
accomplished by chlorination, ozonisation, other chemical disinfectants, UV radiation, membrane processes, or other processes.

Table 14 Concentrations of MSL-treated efuent

NO3 -N (g mL1 )

Phosphate (g mL1 )

Fecal coliform (cfu/100 mL)

MayOctober 2005
Leilehua MSL system
Perlite MSL system

2.15 3.25
3.81 5.35

0.46 0.56
2.16 1.72

658 1321
459 674

MayJuly 2006b
Leilehua MSL system
Perlite MSL system

2.48 2.68
5.04 9.24

2.83 1.38
5.25 2.74

64 95
36 53

a
b

First phase of data without aeration and sucrose addition (mean S.D., n = 21).
Second phase of data with different rates of aeration and constant rate of sucrose, considered as the optimal management of the system
(mean S.D., n = 9).

treated efuent meets the criteria of nitrate and fecal coliform

(MayJuly 2006) of R-2 water and approaches the criteria for R1 water. Improvements in efciency of the type examined in
this study are needed to meet the phosphate criteria. In addition a process, such as chlorination is needed to disinfect the
treated efuent.

5.

The MSL-treated efuent approaches R-1 water criteria, with

improvements in P removal still needed.

Acknowledgement
We gratefully acknowledge the USDA T-STAR Program, University of Hawaii for the support of this research.

Conclusions

Results of this study showed that both MSL systems have the
potential to remediate dairy efuent. The percentage removal
of inorganic N was high and similar in both the MSL systems.
The percentage removal of phosphate was high to very high
in the Leilehua MSL system and it removed considerably more
phosphate than the Perlite MSL system. The supplemental
aeration, which was applied in the second phase of the study,
did not signicantly improve the removal of inorganic N. The
removal of phosphate, however, increased in the Leilehua MSL
system with additional aeration. Application of sucrose with
constant aeration was crucial for removing inorganic N and
phosphate. It appears that sucrose additions increased the
microbial activity in the MSL systems which helped to increase
the removal of inorganic N and phosphate. The sucrose applications have the potential to improve MSL systems treatment
efciency. The installation of MSL systems is simple and basically requires only electricity, freshwater, a constant supply
of efuent and a very small amount of land. The materials
used in the system are inexpensive and easily obtainable.

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