Journal of Environmental Management 90 (2009) 730e739

www.elsevier.com/locate/jenvman

Influence of organic waste type and soil structure on the bacterial

filtration rates in unsaturated intact soil columns
M.R. Mosaddeghi a,*, A.A. Mahboubi a, S. Zandsalimi a, A. Unc b
b

a
Department of Soil Science, College of Agriculture, Bu-Ali Sina University, Hamadan 65174, Iran
Plant and Environmental Sciences, New Mexico State University, Box 30003 MSC 3Q, Skeen Hall Room N336, Las Cruces, NM 88003-8003, USA

Received 11 February 2007; received in revised form 18 October 2007; accepted 13 January 2008
Available online 19 March 2008

Abstract
Organic wastes are considered to be a source for the potentially pathogenic microorganisms found in surface and sub-surface water resources.
Following their release from the organic waste matrix, bacteria often infiltrate into soil and may be transported to significant depths contaminating aquifers. We investigated the influence of soil texture and structure and most importantly the organic waste properties on the transport and
filtration coefficients of Escherichia coli and total bacteria in undisturbed soil columns. Intact soil columns (diameter 16 cm and height 25 cm)
were collected from two soils: sandy clay loam (SCL) and loamy sand (LS) in Hamadan, western Iran. The cores were amended with cow manure, poultry manure and sewage sludge at a rate of 10 Mg ha1 (dry basis). The amended soil cores were leached at a steady-state flux of
4.8 cm h1 (i.e. 0.12 of saturated hydraulic conductivity of the SCL) to a total volume of up to 4 times the pore volume of the columns.
The influent (C0) and effluent (C ) were sampled at similar time intervals during the experiments and bacterial concentrations were measured
by the plate count method. Cumulative numbers of the leached bacteria, filtration coefficient (lf), and relative adsorption index (SR) were calculated. The preferential pathways and stable structure of the SCL facilitated the rapid transport and early appearance of the bacteria in the
effluent. The LS filtered more bacteria when compared with the SCL. The effluent contamination of poultry manure-treated columns was greater
than the cow manure- and sewage sludge-treated ones. The difference between cow manure and sewage sludge was negligible. The lf and SR
values for E. coli and total bacteria were greater in the LS than in the SCL. This indicates a predominant role for the physical pore-obstruction
filtration mechanisms as present in the poorly structured LS vs. the retention at adsorptive sites (chemical filtration) more likely in the better
structured SCL. While the results confirmed the significant role of soil structure and preferential (macroporous) pathways, manure type was
proven to have a major role in determining the maximum penetration risk of bacteria by governing filtration of bacteria. Thus while the numbers
of bacteria in waste may be of significance for shallow aquifers, the type of waste may determine the risk for microbial contamination of deep
aquifers.
2008 Elsevier Ltd. All rights reserved.
Keywords: Adsorption; E. coli; Breakthrough curve; Filtration; Waste; Preferential flow; Soil structure

1. Introduction
Livestock manures have been long considered as beneficial
amendments that enhance soil physical and chemical conditions relevant for plant growth. However, they also release
bacteria such as strains and variants of Salmonella, Shigella

* Corresponding author. Tel.: 98 811 4223367; fax: 98 811 4227012.

E-mail address: mosaddeghi@basu.ac.ir (M.R. Mosaddeghi).
0301-4797/$ - see front matter 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jenvman.2008.01.009

and Escherichia coli which are important human pathogens

(Landry and Wolfe, 1999; Mawdsley et al., 1995). In general,
from more than 1010 bacteria found in 1 g of (dry) manure,
pathogenic bacteria might represent a very small percentage
rarely exceeding 105 bacteria per gram of (dry) manure (Unc
and Goss, 2004).
The fate and survival of organic waste bacteria in soils can
be affected in three ways: (1) adsorption to soils, (2) transport
through soils to the deeper layers and/or towards groundwater,
and (3) die-off due to unfavorable environmental conditions.

M.R. Mosaddeghi et al. / Journal of Environmental Management 90 (2009) 730e739

Many factors may modify these including type of organic

waste, soil pore and particle size distribution, soil permeability, acidity and ionic strength of soil solution, water flow characteristics, climatic conditions and management practices
(Schafer et al., 1995; Gagliardi and Karns, 2000; Unc and
Goss, 2003).
In heterogeneous media such as soils, bacterial transport is
directly related to soil particle size distribution and texture
(Gerba and Bitton, 1984; Tate, 1978). Bacterial cells tend to
flocculate and associate with charged surfaces, and only few
cells remain suspended in the soil solution. Thus bacteria are
adsorbed by soil and organic waste colloids and form colloidebacterial complexes which may easily create bridges
between soil particles that clog soil pores and prevent the infiltration of other cells (Abu-Ashour et al., 1994, 1998).
Soil pore size distribution may clearly affect transport of
bacteria. Under conditions of tillage disturbance, coarsetextured soils have more macropores, and a low adsorption
capacity as well. Therefore, under certain disturbed soil conditions, bacterial transport may be facilitated by coarse-textured
soils (Tan et al., 1992). However, for more structured soils, the
preferential flow due to continuous macropores and cracks is
the main conduit for rapid downward movement of bacteria.
Experimental observations indicated that preferential flow is
the rule rather than the exception in most structured soils
(Flury et al., 1994), and continuous pores that are several times
larger than a bacterium allow bacterial transport over significant distances.
Hagedorn et al. (1978) reported bacterial penetration depth
of 1e83 m through the soil profile depending on soil permeability, degree of saturation and duration of manure leaching.
Continuity of macropores generates a free path for bacterial
movement in intact soil columns (Abu-Ashour et al., 1998;
McMurry et al., 1998; Paterson et al., 1993; Reddy et al.,
1981) and it is generally believed that bacterial transport to
be greater and faster in undisturbed soils as compared to tillage-disturbed soils (Smith et al., 1985). Darnault et al.
(2004) confirmed that in undisturbed soil columns, preferential
flow paths had an important role in the transport of Cryptosporidium parvum. Unc and Goss (2004) investigated the movement of bacteria through sandy loam and silt loam soils in
the field. The results revealed that soil water and bacterial
velocities were higher in the silt loam soil. The difference
was related to aggregation, structural stability and macropores
enhancing preferential flow in the soil with the greater clay
content. Clogging of soil pores depends on pore size, soil texture, water velocity and bacterial dimensions (Stevik et al.,
1999). In disturbed soils, when bacterial cell size was at least
5% larger than the particle size, the pores could filtrate and entrap bacteria (Warnemuende and Kanwar, 2002).
Filtration of manure-borne bacteria in soils depends on soil
and/or manure properties, surface electrochemical characteristics of bacteria and possibly bacterial shape (Unc and Goss,
2003). Organic wastes might affect soil conditions and bacterial transport (Unc and Goss, 2004). Soluble organic compounds may compete with bacteria for retention at positively
charged soil surface loci. This reduces the surfaces available

731

for bacterial sorption and enhances bacterial repulsion (Hall

et al., 2005; Unc and Goss, 2004). Soluble organic substances
in the organic waste suspensions diminish water repellency of
bacteria and might enhance suspension of otherwise hydrophobic bacteria; similarly adsorption onto the surface of suspended organic materials will facilitate bacterial transport
(Unc and Goss, 2004). Shelton et al. (2003) found that in intact
soil columns, the velocity of bacterial plume could be 10 times
higher than the pore water velocity. The large bacterial flux occurred in the presence of organic particulates. Moreover, bacterial survival may be extended in the presence of manure
suspensions (Bradford et al., 2006a,b). Conversely, suspended
organic particulates might clog the free pathways and reduce
the bacterial transport. Darnault et al. (2004) reported that application of cow manure to soil effectively decreased bacterial
flux due to blockage of transport pathways by organic particles. Therefore, bacterial transport studies conducted in the absence of organic waste components may not accurately
characterize the transport potential of bacteria in organic
waste-contaminated environments.
The objectives of this project were to: (i) investigate the influence of soil structure and especially type of organic waste
on bacterial filtration and contamination of effluent from intact
soil columns under unsaturated flow condition, and (ii) identify the factors of significance that may associate with the
risk for bacterial contamination of sub-surface water resources
located at variable depths.
2. Materials and methods
2.1. Soils and sampling procedure
The study was conducted in laboratory using intact soil columns of 16 cm diameter and 25 cm height under unsaturated
steady-state flow conditions. Eighteen undisturbed columns
from each soil were obtained from the 10e35 cm layer by using galvanized metallic core samplers with 3 mm wall thickness, 16 cm internal diameter and a height of 32 cm. Two
soils were sampled: (1) a sandy clay loam (SCL) soil from
the Feiz-Abad area, classified as Typic Xerocrepts, and (2)
a loamy sand (LS) soil from the Cheshme-Ghasaban area,
classified as Typic Xerortents (Soil Survey Staff, 1998), both
located in Hamadan Province. The region has a semi-arid climate, with long-term average annual precipitation of 328 mm.
The internal walls of the samplers were lubricated with liquid paraffin before sampling. The 0e10 cm soil layer was
carefully scraped before sampling to eliminate the effect of
natural surface residue and to include both topsoil (Horizon
A) and subsoil (Horizon B) in the sampled columns. In order
to avoid soil disturbance during sampling, the soil was deeply
watered. Then, the core samplers were pushed gently by hand
or, where required, by using a wooden hammer, into the soil.
The core samples were carefully excavated. The water content,
near the plastic limit, minimized the soil structural disturbance. The soil columns were sealed at the base with metal
screens, brought to the lab and preserved from drying. Disturbed and small undisturbed samples were also taken from

M.R. Mosaddeghi et al. / Journal of Environmental Management 90 (2009) 730e739

732

Table 1
Physical properties of the studied soils (average for 10e35 cm layer)
Sand (g g1) Silt (g g1) Clay (g g1) BD (Mg m3) PD (Mg m3) TP (cm3 cm3) Macro-P (cm3 cm3) Micro-P (cm3 cm3) MWD (mm) Ks (cm h1)
Sandy clay loam soil
0.55
0.22

0.23

1.13

2.63

0.570

0.160

0.410

3.77

39.5

Loamy sand soil

0.84
0.08

0.08

1.46

2.60

0.441

0.143

0.298

2.33

70.6

BD bulk density; PD particle density; TP total porosity; Macro-P macro-porosity; Micro-P micro-porosity, MWD mean weight diameter of soil aggregates, Ks saturated hydraulic conductivity.

the topsoil and subsoil for the purpose of routine lab

measurements.
Particle size distribution was determined using the hydrometer method. Particle density was measured by the pycnometer
method. Bulk density was measured on the undisturbed soil
cores with 5 cm diameter and 7.5 cm height. Total porosity
was calculated from bulk density and particle density. Pore
size distribution was determined from the water retention
curve obtained with the pressure plate method. The relative
volume of the pores with effective diameter greater than
30 mm was defined as Macro-P. Micro-P was determined by
calculating the relative volume of pores with effective diameter smaller than 30 mm. Mean weight diameter (MWD) of soil
aggregates was determined by the wet sieving method. Saturated hydraulic conductivity (Ks) was measured by the constant-head procedure (Klute, 1986). The relevant physical
properties of the studied soils are given in Table 1.
Soil electrical conductivity (EC) and reaction (pH) were determined by EC-meter and pH-meter in 1:5 soil:water suspension and saturation paste, respectively. Soluble potassium and
sodium were determined with a flame photometer, and soluble
calcium and magnesium were measured by the EDTA titration
method in 1:5 soil:water suspension. Calcium carbonate content was measured using the back-titration method. Total organic carbon and dissolved organic carbon (DOC, in 1:5
soil:water suspension) were determined using the wet-digestion method. Cation exchange capacity was determined by
the ammonium-saturation and exchange method (Page et al.,
1992). The chemical properties of the soils are presented in
Table 2.
2.2. Unsaturated steady-state flow conditions
A funnel was mounted and sealed beneath each soil column. The soil columns were placed on tripods in a vertical position. They were saturated with tap water from the base for 2

days, to avoid air entrapment and slaking of the soil surface.

Then, Ks of the columns was measured by the constant-head
method (Klute, 1986).
In order to create similar flow rates on both soils, a constant
flux rate of 4.8 cm h1 (i.e. 0.12 of Ks of SCL) was applied on
the all soil columns. To avoid saturation of the lower boundary
of the columns and to insure steady-state flow condition, a suction was applied using a vacuum pump under the columns. By
initially measuring leaching rates at different suctions, the applied negative pressure was adjusted such that the inlet and
outlet fluxes were equal. Details of the established unsaturated
flow conditions for both soil types are given in Table 3. Waterfilled pore volume (PV) was calculated by multiplying soil
core volume and volumetric water content (measured on five
out of 18 columns). Photo of the experimental set-up is shown
in Fig. 1.
2.3. Sources of faecal indicator bacteria
Cow manure, poultry manure and sewage sludge were used.
Fresh cow manure was collected from traditional cowsheds in
the area of Sanandaj city, western Iran. The cows were fed on
hay and dried bread. Fresh poultry manure was obtained from
the wooden boxes in which free range chickens are sheltered at
night; chickens feed on rice and wheat seeds as well as on insects. Sewage sludge was collected from the wastewater treatment facility of Ghorveh city in western Iran. The sewage
treatment consists simply of the passage of sewage through
successive sedimentation pools without the use of chemical
or physical agents. Sewage sludge was collected from the bottom of these sedimentation ponds. After air-drying, samples
were screened through a 3-mm mesh to obtain uniform subsamples. Organic waste properties were determined by the
same methods described for the soils in Section 2.1. EC, pH,
DOC and soluble cations were determined in 1:20 waste:water
suspension. The water content of the sub-samples was

Table 2
Chemical properties of the studied soils (average for 10e35 cm layer)
pH

SAR

CaCO3
(g 100 g1)

OC
(g 100 g1)

DOC
(g 100 g1)

CEC
(cmolc kg1)

K
(mg L1)

Na
(mg L1)

Mg2
(mg L1)

Ca2
(mg L1)

Sandy clay loam soil

11.40
0.175

7.5

0.33

21.0

0.98

0.09

17.3

16.3

10.9

21.6

60.0

Loamy sand soil

7.80
0.120

7.5

0.13

4.0

0.81

0.36

10.3

6.3

4.0

21.6

48.0

I
(mmol L1)

EC
(dS m1)

I ionic strength of soil solution (I 0.013EC1:1); EC electrical conductivity; pH acidity; SAR sodium adsorption ratio; CaCO3 calcium carbonate content; OC organic carbon content; DOC dissolved organic carbon content; CEC cation exchange capacity; K, Na, Mg2 and Ca2 are soluble cations.

M.R. Mosaddeghi et al. / Journal of Environmental Management 90 (2009) 730e739

Table 3
Characteristics of the established unsaturated flow conditions
Characteristics
Soil water content (g g1)
Soil water content (cm3 cm3)
Suction under soil columns (kPa)
Steady-state flux rate (cm h1)
Apparent pore water velocity (cm h1)
Soil column volume (cm3)
Pore volume (cm3)

Soil
Sandy clay loam

Loamy sand

0.300
0.342
6
4.8
14.0
5025
1720

0.208
0.311
4
4.8
15.4
5025
1557

determined gravimetrically. Relevant chemical properties of

the organic wastes are presented in Table 4. The air-dried organic wastes were applied at a rate of 10 Mg ha1 (dry basis)
on top of the soil columns.
2.4. Bacteria selection
E. coli is used as an indicator of faecal contamination. E.
coli is a Gram-negative, mobile, aerobic or facultative anaerobic bacterium with cell diameter of 1e6 mm (Jawetz et al.,
2001). The waste-borne E. coli and total bacterial concentrations were measured in both the columns influent and effluent
for all treatments. Here total bacteria refers to the Gramnegative facultative anaerobic and aerobic Enterobacteriaceae
that were recovered on Eosin Methylene Blue (EMB) media at
37  C for 18e24 h.
Initial concentrations of E. coli and total bacteria were also
determined in the waste suspension. The suspensions were
shaken manually, extracted through a thin layer of sterile cotton, diluted as necessary and sub-samples of 0.1 mL were
cultured on EMB plates. Plates were incubated at 37  C for
18e24 h. Differential growth media, the IMVIC test (Jawetz
et al., 2001), was also used to differentiate E. coli from other

733

Gram-negative bacteria. This test included the use of Urea

Agar, Citrate Agar, Lysine, Triple Sugar Iron Agar, Sulfide Indole Motility test, and Methyl Red and VogeseProskauer Reactions (Jawetz et al., 2001). Results were reported as colony
forming units (CFU) per milligram of dry waste. The average
concentration of E. coli in the wastes was 45  104, 88  102
and 25  102 CFU mg1 for poultry manure, sewage sludge
and cow manure, respectively. The same order was observed
for the total bacteria concentrations with average values of
48  104, 39  103 and 35  103CFU mg1, respectively.
Therefore, poultry manure is a source of significantly more
bacteria.

2.5. Leaching experiments

After establishing the steady-state flow condition, organic
wastes were spread homogeneously on the soil columns
(20.1 g dry basis for each column) and the leaching experiment started. The waste-amended soil columns were leached
with up to 4 PVs of water over a period of about 6.5 h, at
20  C. Same laboratory conditions were maintained for all
treatments. The effluent was sampled at 0.1, 0.2, 0.3, 0.4,
0.6, 0.8, 1, 1.3, 1.6, 1.9, 2.2, 2.6, 3, 3.5 and 4 PVs by sterilized
medical needles to determine effluent concentration (C ). In order to monitor the background concentration (Cb) for E. coli
and total bacteria, one undisturbed column of each soil was
leached under the same flow condition and effluent was sampled at the same intervals as for the treated columns. The
background E. coli was negligible but the background total
bacteria in the LS and SCL columns were estimated at
18  106 and 78  106 CFU per column, respectively.
In order to estimate the influent concentration (C0), similar
quantities of air-dried waste were placed over a sterile mesh
basket and leached with the same water flux rates as used

Fig. 1. Photo of leaching experimental set-up.

734

M.R. Mosaddeghi et al. / Journal of Environmental Management 90 (2009) 730e739

Table 4
Chemical properties of the tested wastes
Property

Cow
manure

Poultry
manure

Sewage
sludge

Electrical conductivity (dS m1)

Ionic strength, I 0.013EC1:1 (mmol L1)
Acidity (pH)
Sodium adsorption ratio (SAR)
Gravimetric water content (%)
Organic matter content (%)
Dissolved organic carbon content (%)
Soluble potassium (mg L1)
Soluble sodium (mg L1)
Soluble magnesium (mg L1)
Soluble calcium (mg L1)

3.00
780.0
8.4
2.5
8.1
77.7
5.4
4000
144.6
57.6
168

2.10
546.0
7.0
7.0
7.7
76.4
3.6
1950
128.9
43.2
720

0.26
67.6
7.7
7.7
6.8
24.6
0.72
7.8
4.4
100.8
96

columns. Growth/die-off parameters are also non-explicit

and thus are intrinsically integrated in the final results; however, given the uniform experimental conditions, die-off and
growth parameters are expected to be rather similar. It is difficult to differentiate physical straining and chemical adsorption. So relative adsorption index (SR) was calculated as
a relative quantity of bacteria retained in the soil columns during the leaching experiments by:
ZVmax
SR

C0  C  Cb dV

ZVmax
C0 dV
0

for the soil column experiments. Leachate was sampled at the

same intervals as for the column experiments. Concentrations
of E. coli and total bacteria in the samples were obtained by
the plate count method as described in Section 2.4 and the results were reported as CFU mL1.

where Vmax is the maximum effluent volume (mL). In Eq. (2),

the surface area between the influent curve and BTCs (after
subtraction of background concentration) was divided by the
surface area under the influent curve; both were calculated
by the mentioned algorithm.

2.6. Statistical design and data analysis

3. Results and discussion
Breakthrough curves (BTCs) of untreated and treated columns, and for influent concentrations of E. coli and total bacteria were drawn as Cb vs. PV, C vs. PV, and C0 vs. PV. The
experimental design was a two-factor factorial complete randomized blocks with three replicates. The treatments combined soil type (SCL or LS) and waste type (cow manure,
poultry manure or sewage sludge). Thus 24 waste-treated
soil columns and two untreated soil columns (in total 26)
were leached. The dependent variables were the cumulative
number of E. coli and total bacteria leached during the experiment, the calculated filtration coefficient (lf), and relative adsorption index (SR). Data were analyzed using SAS and the
mean comparisons were done by LSD (SAS Institute, 1990).
Cumulative numbers of the passed bacteria were calculated
by using the simulation algorithm in MATLAB code for the
same cumulated effluent volume (i.e. four PVs of LS). This
was done by calculating the surface area under the BTC after
subtraction of background bacteria (Cb).
The filtration coefficient (lf) was calculated according to
Mathess et al. (1988):


C0av
1

1
lf ln
Cav  Cbav
x
where lf is expressed in m1, C0(av), C(av) and Cb(av) are the
average influent, effluent and background concentrations
(CFU mL1), and x is the length of the soil column (m). In
order to determine C0(av), C(av) and Cb(av), the surface areas
below the influent curve and BTCs of treated and untreated
soil columns were calculated by using the mentioned algorithm and divided by the cumulative effluent volume of 4 PV.
The difference between the influent and effluent concentrations (after subtraction of background concentration) equals to
the bacteria that are filtered and/or adsorbed within the soil

3.1. Breakthrough curves of E. coli and total bacteria

BTCs of E. coli and total bacteria are illustrated in Figs. 2
and 3, respectively. The C0 variation during leaching was also
monitored. The C0 curves were above the BTCs showing filtering effects of the soils. In general, LS columns better filtered
bacteria due to weaker structure and discontinuity of pores.
The greater clay content (Table 1) in the SCL facilitated bacterial transport and early appearance of bacteria in the leachate
likely due to enhanced structure stability and continuity of the
macropores. Bacterial concentration of the effluent from poultry manure-treated columns was greater than the cow manureand sewage sludge-treated ones. The difference between cow
manure and sewage sludge was negligible.
The cow manure had a lot of decay-resistant straws consisting of lignin and cellulose, which are not suitable energy sources for Gram-negative bacteria such as E. coli. During initial
leaching of cow manure, C of the LS columns was greater
than for the SCL columns. When leaching continued beyond
0.3 PV, C of the LS decreased due to physical filtration
(Figs. 2a and 3a). However, C of the SCL had an increasing
trend during initial leaching. Parallel to decrease of C0 during
leaching, C also gradually decreased indicating a direct effect
of bacterial release from cow manure on bacteria leaching
from this soil (Figs. 2a and 3a).
The C values of poultry manure-treated columns were the
highest among all wastes (Figs. 2b and 3b). The fast movement
of bacteria along the macropores of the SCL columns diminished
the waste effects. As leaching continued (after 0.2 PV), the differences between C of the two soils diminished. Soil pore clogging
by suspended materials of poultry manure is suspected to have
caused bacterial filtration in the LS columns. Towards the end,
C increased due to partially delayed breakthrough.

M.R. Mosaddeghi et al. / Journal of Environmental Management 90 (2009) 730e739

7.5
7
6.5
6
5.5
5
4.5
4
3.5
3
2.5

a
LS
SCL
C0

0.5

Log Concentration
(CFU ml-1)

Log Concentration
(CFU ml-1)

1.5

2.5

3.5

LS
SCL
C0
0

0.5

1.5

2.5

3.5

4.5

Log Concentration
(CFU ml-1)

7.5
7
6.5
6
5.5
5
4.5
4
3.5
3
2.5

0.5

LS
SCL
C0

1.5

2.5

3.5

4.5

7.5
7
6.5
6
5.5
5
4.5
4
3.5
3
2.5

LS
SCL
C0
0

0.5

1.5

2.5

3.5

4.5

Pore volume (V/V)

c
Log Concentration
(CFU ml-1)

Pore volume (V/V)

Pore volume (V/V)

Log Concentration
(CFU ml-1)

Log Concentration
(CFU ml-1)

7.5
7
6.5
6
5.5
5
4.5
4
3.5
3
2.5

LS
SCL
C0

4.5

Pore volume (V/V)

7.5
7
6.5
6
5.5
5
4.5
4
3.5
3
2.5

735

7.5
7
6.5
6
5.5
5
4.5
4
3.5
3
2.5

LS
SCL
C0

0.5

1.5

2.5

3.5

4.5

Pore volume (V/V)

0

0.5

1.5

2.5

3.5

4.5

Pore volume (V/V)

Fig. 2. Breakthrough curves of E. coli concentrations released from different
manures and manure-treated soil columns: (a) cow manure, (b) poultry manure
and (c) sewage sludge. LS, SCL and C0 stand for loamy sand soil, sandy clay
loam soil and inlet concentration of E. coli, respectively.

Sewage sludge is a diluted and anaerobic medium. The C of

sewage sludge-treated LS columns was high when leaching
started (Figs. 2c and 3c). The C quickly decreased in parallel
with the decrease of C0. The trend was different for the SCL
columns; the C initially increased and, after 0.3 PV, gradually
decreased. Overall, little filtration occurred when sewage
sludge was used (Fig. 3c).
When the LS was leached, the C of bacteria was initially
lower in poultry manure treatment (Figs. 2 and 3). When leaching started, low ionic strength of sewage sludge suspension (Table 4) caused fast movement of bacteria towards the bottom of
the columns. Thus the C of sewage sludge-treated LS columns
was greater than cow manure-treated ones. However, when
leaching continued the reverse trend was observed, i.e. more
bacteria were retained in the soil due to pore clogging by suspended organic particulates of cow manure. In general, the cow
manure had greater effects on the transport of bacteria through

Fig. 3. Breakthrough curves of total bacterial concentrations released from different manures and manure-treated soil columns: (a) cow manure, (b) poultry
manure and (c) sewage sludge. LS, SCL and C0 stand for loamy sand soil,
sandy clay loam soil and inlet concentration of total bacteria, respectively.

the soil correlated to its high ionic strength, soluble cations, and
soluble and suspended organic particulates (Table 4).
The significance of wastes on bacterial filtration in the SCL
was diminished because of the dominant impact of macroporous pathways (Figs. 2 and 3). The early appearance of bacteria in the effluent was obvious for poultry manure-treated SCL
columns (Figs. 2b and 3b). The C of poultry manure-treated
SCL columns had an initially increasing trend and after 0.3
PV, it varied little. The C of sewage sludge-treated columns
considerably decreased after 0.3 PV. Sewage sludge had low
ionic strength and soluble and/or suspended organics so that
it quickly released the bacteria.
3.2. Cumulative number of leached E. coli and total
bacteria
The influences of soil type and organic waste type, and
their interactions on cumulative number of E. coli and total
bacteria passing through the soil columns were significant

736

M.R. Mosaddeghi et al. / Journal of Environmental Management 90 (2009) 730e739

Table 5
Mean comparisons of cumulative number (CFU) of leached E. coli and total bacteria as influenced by soil and manure treatments
Soil

E. coli
Sandy clay loam
Loamy sand
X
LSD (0.05)
Total bacteria
Sandy clay loam
Loamy sand
X
LSD (0.05)

Manure
Cow manure

Poultry manure

Sewage sludge

28  107 (81  105)

58  107 (17  105)
43  107
Soil (S )

72  109 (17  107)

38  109 (12  107)
55  109
Manure (M )

79  106 (9  105)

49  107 (15  105)
28  107
SM

17  108

16  108

11  108

49  107 (17  107)

31  107 (79  106)
40  107
Soil (S )

19  109 (10  109)

98  108 (28  108)
14  109
Manure (M )

37  107 (12  107)

40  107 (12  107)
39  107
SM

13  108

49  108

27  108

24  109
13  109

66  108
35  108

Figures in the parentheses are the standard deviations.

(P < 0.05). The total number of leached bacteria (Table 5) is

a suitable and reliable index to examine the impacts of the factors on bacterial transport in comparison to the instantaneous
concentrations (shown in Figs. 2 and 3).
There were significant differences between the means of E.
coli and total bacteria cumulative numbers for the two soil
types (Table 5). Despite the larger apparent adsorptive capacity (clay and organic particles) of the SCL, the LS filtered out
1.86 times more bacteria than the SCL. The greater Macro-P
(Table 1) and continuity of the macropores in the SCL facilitated bacterial transport. This implies that even under unsaturated conditions, preferential flow might be of significance.
Furthermore it seems that in the unsaturated fine-textured
soil, water films and menisci are mostly connected and effectively transmit bacteria. Smith et al. (1985) found that the
main way of bacterial transport in intact structured soils is
preferential flow. Abu-Ashour et al. (1998), Fontes et al.
(1991), Gerba and Bitton (1984), McMurry et al. (1998), Paterson et al. (1993) and Tate (1978) reported that the spatial
distribution and continuity of macropores (i.e. preferential
pathways) are the main means of bacterial transport in unsaturated structured soils.
The type of organic waste significantly affected the cumulative bacterial transport through the soil columns. The mean cumulative numbers of leached E. coli and total bacteria were in
the order: poultry manure > cow manure > sewage sludge
(Table 5). The relative magnitudes of number of leached
E. coli were 196.4:1.5:1 for poultry manure, cow manure and
sewage sludge, respectively. This means that, for the total
length of these soil cores, the effluent pollution caused by poultry manure was 196.4 and 130.9 times higher than the one
caused by the sewage sludge and cow manure, respectively.
The high C from the poultry manure treatment was directly
related to the great number of bacteria released from this
manure. The total numbers of E. coli released from the poultry
manure during leaching were 241.9 and 199.4 times higher than
sewage sludge and cow manure, respectively. The difference
between poultry manure and the other two organic wastes
was significant in terms of the cumulative number of leached

bacteria, but the difference was not significant between cow manure and sewage sludge (Table 5). Bradford et al. (2006a) compared the somatic Coliphage transport in the presence and
absence of manure suspension. The results revealed the survival
of pathogen and much higher effluent concentrations in the
presence of manure. The difference was attributed to lower inactivation and higher detachment rates. While inactivation
mechanisms for bacteria are different from the ones for Coliphages, the higher detachment rates in the presence of manure
components may be of significance for both groups of
microorganisms.
The suspended organic particulates offer appropriate loci
for bacterial sorption which eventually enhance bacterial
movement (Unc and Goss, 2003). Conversely, suspended organic particulates might clog the bacteria-transmitting pores
and capture more bacteria within the soil. The interactive effect on the effluent pollution was significant (Table 5). It appears that the organic waste effect on bacterial transport was
minimized in the SCL columns due to dominant role of bypass
flow. However, bacterial transport was retarded in the LS for
all organic waste treatments.
3.3. Bacterial filtration coefficient
Organic waste type had a significant impact on the filtration
coefficient (lf) for E. coli. However, the soil type had a significant effect on lf for total bacteria (Table 6). It is presumed
that variations in cell properties resulted in significant differences in the transport phenomena among microorganisms.
The mean lf values of E. coli and total bacteria for the LS
were 1.2 and 1.3 times higher than the corresponding values
for the SCL, respectively. However, the differences between
soils were only significant for the filtration coefficients of total
bacteria (Table 6). Generally, the LS retained more bacteria,
which is in agreement with Unc and Goss (2003). Foppen
et al. (2005) also found that straining in dead-end pores was
an important process in fine-grained sediment. For the structured SCL, the water velocity is very high in the cracks and
macropores which would limit the bacterial adsorption or

M.R. Mosaddeghi et al. / Journal of Environmental Management 90 (2009) 730e739

Table 6
Mean comparisons of filtration coefficient (lf, m1) of E. coli and total bacteria as influenced by soil and manure treatments
Soil

E. coli
Sandy clay loam
Loamy sand
X
LSD (0.05)
Total bacteria
Sandy clay loam
Loamy sand
X
LSD (0.05)

Manure
Cow
manure

Poultry
manure

Sewage
sludge

0.35 (0.38)
1.06 (0.13)
0.71
Soil (S )

2.37 (0.44)
3.17 (1.02)
2.77
Manure (M )

2.22 (1.14)
1.63 (0.40)
1.93
SM

0.96

1.54

1.79

2.26 (1.31)
3.40 (1.12)
2.83
Soil (S )

2.32 (0.42)
3.09 (1.1)
2.71
Manure (M )

2.06 (1.29)
2.36 (1.16)
2.21
SM

0.68

1.85

2.16

737

lowest lf value for E. coli. For the total bacteria filtration,

the highest and the lowest values were calculated for the
LSepoultry manure and the SCLesewage sludge combinations (Table 6).
3.4. Relative adsorption index for E. coli and total
bacteria

1.65
1.95

2.21
2.95

Figures in the parentheses are the standard deviations.

filtration in the fine pores. Therefore, the overall filtration was

interestingly lower in the soil with more total adsorptive sites
(i.e. SCL). This could also be inferred from the structural stability results (i.e. MWD) of the soils (Table 1), indicating that,
for particulate retention, the soil adsorptive capacity needs to
be evaluated in the context of the pore sizes available for particulate penetration and transport. Another reason might be the
continuity of the water films and menisci around the soil particles in the finer soil.
The properties of the bacteria-carrying suspension have
a great effect on the bacterial filtration. Gagliardi and Karns
(2000), Johnson and Logan (1996) and Unc and Goss (2003)
observed that presence of manure components might decrease
bacterial filtration through soil columns. Bradford et al.
(2006b) also reported higher effluent concentrations of Giardia in the presence of manure suspension. In our case, some
different results were obtained concerning the organic wasteinduced bacterial transport. The mean lf values for E. coli
were in descending order: poultry manure > sewage sludge >
cow manure. The ionic strength of poultry manure suspension
was higher than that of sewage sludge (Table 4) which favors
bacterial flocculation and retention to soil surfaces by minimizing the electrical repulsive effect of same-charged particles
(Unc and Goss, 2004). However, the same logic does not apply
when the comparison includes the filtration coefficient of
E. coli for the cow manure (Table 6). Nevertheless, lf was
inversely correlated to the DOC concentration of the three
organic wastes suggesting a role for the soluble carbon compounds in modifying the retention of E. coli in the soil columns. Filtration results were less obvious for the total
bacteria. The ratios of lf values of total bacteria for cow manure, poultry manure and sewage sludge were 1.28:1.23:1.00,
respectively (Table 6).
The interactive effects were significant for lf of E. coli, but
were not significant for total bacteria. The LS columns treated
with the poultry manure filtered the greatest amount of E. coli,
while the SCL columns treated with the cow manure had the

The E. coli and total bacteria adsorption and filtration were

affected by soil and organic waste treatments (Table 7). The
values of relative adsorption index (SR) were higher for the
LS. However, Huysman and Verstraete (1992), Lo et al.
(2002), Stevik et al. (1999) and Tan et al. (1992) all showed
that, in disturbed soil columns, bacterial straining increased
with the increase in the clay content. Working with intact
soil cores, Hekman et al. (1995) also observed that bacterial
filtration was 29% higher in a silt loam soil than in a loamy
sand soil. The bypass and macroporous flow pathways due
to aggregation and inter-aggregate pores as well as the higher
ionic strength of soil solution (Table 2) might be the reasons
for the lower SR values in the SCL. Similar results were recorded for the total bacteria. The LS filtered 1.24 times
more total bacteria than the SCL (Table 7).
There were significant differences between the waste treatments in terms of E. coli retention (Table 7). The mean values
of SR for the wastes were in order: 0.49 > 0.38 > 0.16 for
poultry manure > sewage sludge > cow manure, respectively.
These numbers reveal that 49% of the poultry manure-released
E. coli were retained in the soil columns. The soil columns
treated with the poultry manure filtered 129 and 306% more
E. coli than the sewage sludge- and cow manure-treated soil
columns. The high concentration of the influent as well as
ionic strength of the poultry manure suspension likely enhanced formation of the bacteria/organic matter complexes,
which are several times larger than individual bacteria. Straining and filtration of these complexes are much easier and this
may be the reason for higher relative filtration (SR) in poultry
manure-treated soil columns. The cow manure and sewage
sludge suspensions had lower initial concentrations of E.
Table 7
Mean comparisons of relative adsorption index (SR) of E. coli and total bacteria as influenced by soil and manure treatments
Soil

E. coli
Sandy clay loam
Loamy sand
X
LSD (0.05)
Total bacteria
Sandy clay loam
Loamy sand
X
LSD (0.05)

Manure
Cow manure

Poultry manure

Sewage sludge

0.08 (0.09)
0.23 (0.03)
0.16
Soil (S )

0.44 (0.06)
0.53 (0.13)
0.49
Manure (M )

0.41 (0.17)
0.34 (0.07)
0.38
SM

0.16

0.23

0.29

0.41 (0.20)
0.56 (0.11)
0.49
Soil (S )

0.44 (0.06)
0.53 (0.14)
0.49
Manure (M )

0.38 (0.21)
0.43 (0.17)
0.41
SM

0.04

0.58

0.31

Figures in the parentheses are the standard deviations.

0.31
0.37

0.41
0.51

738

M.R. Mosaddeghi et al. / Journal of Environmental Management 90 (2009) 730e739

coli. However, the colloidal and soluble organic substances of

cow manure (Table 4) facilitated the E. coli transport. Unc and
Goss (2004) stated that organic electrolytes might increase the
adsorption of bacteria on suspended charged organic compounds and consequently enhance bacterial movement. Guber
et al. (2005a,b) found that attachment of E. coli to soil was
much smaller in the presence of manure, and decreased with
the increase in manure contents. They related the reduced attachment to soil and increased survival of E. coli to slow manure colloid transport and high concentration in pore solution.
The results were different for the total bacteria (Table 7). The
relative values of SR for the wastes were 1.20:1.20:1.00 for
cow manure, poultry manure and sewage sludge, respectively.
It means that cow manure- and poultry manure-treated columns retained 1.20 times more total bacteria than sewage
sludge. It may be, however, of interest for latter studies, to
identify the significance of organic wastes in modulating the
numbers of total bacteria and thus their presence in leachate.
The interactive effects of the treatments on SR were not significant (Table 7). The highest values of SR for E. coli and total
bacteria were observed for the poultry manureeLS combination. The lowest SR values of E. coli and total bacteria were
calculated for the SCL columns treated with cow manure
and sewage sludge, respectively.
4. Conclusions
Results demonstrated that the LS filtered more bacteria due
to weaker structure and discontinuous pores. Structural stability
in the SCL facilitated bacterial transport and early appearance
of bacteria in the leachate. The contamination of effluent from
poultry manure-treated columns was greater than the cow
manure- and sewage sludge-treated ones. The effects of wastes
on bacterial filtration in the SCL were lower because of dominant impact of macroporous pathways. The findings illustrated
the significant role of the waste type in determining potential for
deep transport of bacteria. Cow manure although released less
total bacteria, it did prove to increase the maximum penetration
risk of bacteria. Thus, while soil porosity plays a significant role
in the vertical transport of bacterial cells through unsaturated
soils, the type of waste and thus the characteristics of the organic
matter associated with a specific organic waste may also govern
the maximum contamination depth. It is therefore essential to
understand the characteristics of the waste organic matter compounds that may govern bacterial transport through soils to
be able to improve our transport modeling capabilities and
also to target waste treatment strategies.
Acknowledgments
Appreciation is expressed to Bu-Ali Sina University of
Hamadan and Kurdestan Medical Sciences University of Sanandaj for the partial support of the research. The authors also
acknowledge the financial support of Iran National Science
Foundation (INSF) for Project No. 84147 which made the
study possible.

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