1
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Evaluation of biochar soil amendments in
reducing soil and water pollution from total
and fecal coliforms in poultry manure
Shoieb A. Arief Ismail1, Shiv O. Prasher1*, Martin R. Chénier2 and Ramanbhai M. Patel1
Department of Bioresource Engineering, McGill University, Sainte Anne de Bellevue, Quebec H9X 3V9, Canada
2
Department of Animal Science and Department of Food Science, McGill University, Quebec H9X 3V9, Canada
Email: [email protected]
http://dx.doi.org/10.7451/CBE.2016.58.1.21
1
Received: 2016 July 21, Accepted: 2016 September 9, Published: 2016 December 15.
Arief Ismail, S.A., S.O. Prasher, M.R. Chénier and R.M. Patel.
2016. Evaluation of Biochar Soil Amendments in Reducing
Soil and Water Pollution from Total and Fecal Coliforms in
Poultry Manure.
Laboratory and field studies
were conducted to evaluate the adsorption behavior of a biocharamended sandy soil for Escherichia coli (E. coli). Two types of
biochars, produced respectively by either slow pyrolysis (SPB) or
fast pyrolysis (FPB), were used. In the laboratory study,
soil+SPB and soil+FPB treatments (99:1 soil:biochar by weight)
resulted in a significantly higher adsorption after 24 hours as
compared to soil alone (H0: p<0.05). After 7 days, the
concentration of E. coli in solution phase were 0.87±0.08 x 106,
<1.00 x 103, and 1.17±0.15 x 104 in soil alone, soil+SPB and
soil+FPB treatments, respectively. The effectiveness of
soil/biochar amendments was further tested on field lysimeters by
studying the fate and transport of total and fecal (E. coli)
coliforms from poultry manure. Four irrigation events were
simulated. Periodic soil samples at three sampling depths as well
as leachate samples were collected and analyzed. Total coliforms
and E. coli were found to move through the soil profile (0.10 m,
0.31 m and 0.52 m), although their concentrations decreased with
soil depth and time. In biochar-amended lysimeters, the
concentration of total coliforms was significantly lower (H 0: P<
0.05) than in control (soil alone) lysimeters, at most sampling
times. The concentrations of E. coli in the biochar treatments, at
the three soil sampling depths and in the leachate, were also
significantly lower than in the control (H0: P < 0.05). Therefore,
both biochar amendments were effective in reducing leaching of
E. coli through the soil profile and in providing a fecal coliformfree leachate. This study shows that biochar can play an
important role in improving drainage water quality and in
addressing the concerns for pathogen leaching as a result of the
use of poultry manure on agricultural land. Keywords: Biochar;
Total coliform; Fecal coliform; Escherichia coli; Poultry Manure.
Des études en laboratoire et aux champs ont été réalisées
pour évaluer la capacité d’adsorption de l’Escherichia coli (E.
coli) d’un sol sablonneux amendé avec du biochar. Deux types de
biochars ont été utilisés, soit celui produit par une pyrolyse lente
(SPB) et celui produit par une pyrolyse rapide (FPB). Dans
l’étude en laboratoire, les traitements sol+SPB et sol+FPB (ratio
99:1, sol:biochar sur une base massique) ont donné une
adsorption significativement plus élevée après 24 h
comparativement au sol seul (H0 : p < 0,05). Après 7 jours, la
concentration en E. coli de la phase solution était de 0,87 ± 0,08
x 106, <1,00 x 103 et 1,17 ± 0,15 x 104, respectivement pour le
sol seul, le traitement sol+SPB et le traitement sol+FPB.
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L’efficacité des amendements au biochar a aussi été testée aux
champs avec des lysimètres en étudiant les concentrations et le
transport des coliformes totaux et fécaux (E. coli) de fumier de
volailles. Quatre évènements d’irrigation ont été simulés. Des
échantillons périodiques de sol prélevés à 3 profondeurs ainsi que
des échantillons de lixiviat ont été recueillis et analysés. Les
coliformes totaux et E. coli se sont déplacés à travers le profil de
sol (0,10 m, 0,31 m et 0,52 m) toutefois, leurs concentrations ont
diminué avec la profondeur du sol et le temps écoulé. Dans les
lysimètres amendés avec du biochar, la concentration totale de
coliformes était significativement plus faible (H0 : P < 0,05) que
dans le lysimètre témoin (sol seul) la plupart des périodes
d’échantillonnage. Les concentrations en E. coli des traitements
avec du biochar, à trois profondeurs d’échantillonnage et dans le
lixiviat, étaient significativement plus faibles que dans le témoin
(H0 : P < 0,05). Par conséquent, les deux amendements au
biochar réduisaient efficacement la lixiviation de l’E. coli dans le
profil de sol et produisaient un lixiviat sans coliformes fécaux.
Cette étude démontre que le biochar peut jouer un rôle important
en améliorant la qualité des eaux de drainage et en offrant une
solution à la lixiviation de pathogènes résultant de l’épandage de
fumier de volailles sur les terres agricoles. Mots clés: biochar,
coliformes totaux, coliformes fécaux, Escherichia coli, fumier de
volailles.
INTRODUCTION
Poultry facilities harbour and shed bacteria, parasites, and
viruses, some of which are pathogenic to humans,
livestock, domestic animals, and wildlife. Many of these
pathogens are endemic in poultry and are difficult to
eradicate from production facilities. For instance, the
prevalence of Campylobacter jejuni and Salmonella spp.
in poultry operations has been reported to be as high as
100% (Rogers and Haines 2005). A survey of the literature
reveals that poultry manure is a reservoir of
Campylobacter jejuni, Escherichia coli (including
O157:H7), enteropathogenic Escherichia coli, and
Salmonella spp. (non typhi or paratyphi) (Rose et al. 1999;
Bolton et al. 1999; Nicholson et al. 2002; Chevalier et al.
2004; Furtula et al. 2010; Diarra and Malouin 2014;
Atidegla et al. 2016). The colon of poultry can harbour
pathogenic bacteria at levels that can reach millions to
billions per gram of feces. Once excreted in feces,
pathogenic bacteria may survive for weeks to months and
even grow in untreated animal manure and manure
slurries, where such microorganisms benefit from high
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concentrations of nutrients and protection from UV
radiation, desiccation, and temperature fluctuations (ASAE
Standards 2005).
Poultry manure has excellent fertilizing value, and
thus, large amounts are usually applied prior to planting in
the spring, at crop emergence, and to a lesser extent, in the
fall (van Bochove et al. 2010). Given the presence of
pathogens in these manures, surface waters are more
susceptible to being contaminated by coliforms carried in
runoff from agricultural land (Borst and Selvakumar 2003;
Stout et al. 2005; Jenkins et al. 2006; Omarova et al.
2016). Contamination can also occur through subsurface
drainage waters as the pathogens and coliforms can be
transported through soil and water (Pappas et al. 2008;
Megchún-García et al. 2015). The consequences of
contamination of water by pathogens and coliforms
include closure of water bodies for recreational use,
elevated costs for treatment of contaminated water,
interference with the expansion of the livestock industry,
and health issues in animals and humans. Thus, treatment
of surface and subsurface flow from agricultural land as
well as from livestock and poultry industries is necessary.
Groundwater contaminated by sewage or animal
manure is evaluated in terms of the presence of fecal
coliforms, such as E. coli, which is widely recognized as
an “indicator organism”. Its presence is indicative of the
potential presence of pathogens, which cause diseases
(Health Canada 2011). In Canada, the maximum allowable
levels of E. coli in waters for recreational use (e.g.
swimming) and drinking purposes are ≤ 200 CFU dL-1 and
0 CFU dL-1, respectively (Health Canada 2006). Waters
having concentrations above these levels are considered
unsafe for these purposes. Hence, it is important to
disinfect water, containing pathogens before it reaches
water bodies.
Different water treatment methods for households,
such as distillation, ultraviolet light, chlorination,
ozonation, and ceramic candle filtration have been used
(Health Canada 2008; Shanon et al. 2008). However, these
methods are expensive and also cannot be applied to large
volume runoff waters; alternative and inexpensive
methods are necessary. Filtration of runoff water through
soil and other filtering media may provide a better option.
Biochar, the charcoal produced from pyrolysis of
biomass, is gaining global recognition as a soil amendment
due to its unique properties. It reduces leaching of nitrogen
through subsurface flow, reduces emission of nitrous oxide
into the atmosphere, and increases cation-exchange
capacity, thereby, improving soil fertility, moderating soil
acidity, increasing water retention in soil, and favoring
beneficial soil microbes (Pietikäinen et al. 2000; Lehman
et al. 2011; International biochar initiatives, 2016).
Biochar is highly porous and has a large surface area, up to
400 m2 per g, depending on its mode of production and
biomass of origin (Liang et al. 2006; Downie et al. 2009).
Microbial cells (bacteria, fungi, actinomycetes and
lichens) have a size range from 0.5 µm to 5 µm, while
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algal cells can vary from 2 µm to 20 µm (Lal 2006). The
dimensions of biochar macropores are above this range
and could serve as habitats for microbes, which protect
them from predation and desiccation (Thies and Rillig
2009; Saito and Muramoto 2002; Warnock et al. 2007).
Dalahmeh et al. (2012) reported that activated charcoal
reduces E. coli to some extent from greywater, and that
such water could be used for irrigation. Moreover, Moges
et al. (2015) recently reported that biochar was used to
remove total coliforms and E. coli from student dormitory
wastewater. The half-life of biochar is estimated to be
hundreds of years (Roberts et al. 2010) and therefore, it is
possible that its role in reducing agricultural pollution
could be long lasting and cost-effective. Biochar appears
to be a promising candidate for filtering pathogens from
large volumes of agricultural runoff or drainage flows.
However, the effect of biochar amendment on the fate and
transport of coliforms in soil-water systems is not clearly
known. Therefore, in the present work, studies were
conducted in the laboratory to evaluate the potential of
biochar to adsorb E. coli (fecal coliform). Also, a field
study using lysimeters was conducted to investigate its
effectiveness as a soil amendment for the mitigation of the
migration of E. coli (fecal coliform) and total coliforms
through the soil column and leachate.
MATERIALS AND METHODS
Laboratory studies
In order to compare the adsorption potential of a sandy
soil, SPB and FPB, a bacterial adsorption study was
conducted with Escherichia coli (E. coli) strain B (ATCC
# 11303, Cederlane, Burlington, ON, Canada), a biosafety
level 1 microorganism. The E. coli strain was grown in
Nutrient Broth (BD Scientific, USA).
The laboratory study was conducted in triplicate 50
mL sterile centrifuge tubes (Fischer Scientific, SaintLaurent, QC, Canada). Autoclave-sterilized materials in a
sterile biological safety cabinet environment were used to
prevent contamination. Sandy soil was collected from field
lysimeters. The study was conducted with 6 g of either
sandy soil, or SPB-amended sandy soil (99:1 = 5.94 g
sandy soil and 0.06 g SPB; soil+SPB), or FPB-amended
sandy soil (99:1 = 5.94 g sandy soil and 0.06 g FPB;
soil+FPB). Each sterile centrifuge tube was inoculated
with 1 mL of the E. coli culture described above, 30 mL of
autoclaved-sterilized deionized water were added, and
adsorption experiment was carried out at 28°C in a
controlled environment in a shaker (Max Q 4000, Thermo
Scientific, USA) operating at 300 rpm for 7 days.
The enumeration of E. coli was done by the
membrane filtration method using US EPA Method 10029
(APHA-AWWA-WEF 1998). A buffer solution for the
dilution of supernatant samples was prepared by adding
1.25 mL of stock phosphate buffer solution (0.25 M
KH2PO4 adjusted to pH 7.2 with 1 N NaOH) and 5.0 mL
of 0.4 M MgCl solution to 1 litre of deionized water. The
buffer solution was autoclaved for 15 min at 1300C and
stored in aliquots for dilution. Following the adsorption
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experiment, a 1 mL aliquot was aseptically collected from
each triplicate after 24 h, 3 days and 7 days. Under sterile
conditions, sample supernatants were diluted so that one
would expect 20 to 200 CFUs to be retained per filter.
Supernatant samples (1 mL) were each diluted in 9 mL of
buffer solution (1:10 dilution) and were subjected to
further serial dilutions (up to 10-7). Samples and dilutions
(10 mL each) were individually vacuum-filtered under
sterile conditions, and the bacteria-retaining filter (0.45 µm
pore size, 47 mm diameter) was rinsed with 30 mL of
sterile buffer. The filter was placed in a prepared, sterile
Petri dish containing liquid broth Hach m-Coli Blue 24,
evenly saturated into an adsorbent pad. The Petri dish was
inverted and incubated for 24 h at 35±0.5o C. The colonies
were counted under 10-15X magnification. E. coli (fecal
coliform) grew as blue colonies whereas total coliforms
grew as red colonies. A few varieties of non-coliform
bacteria (Pseudomonas, Vibrio, and Aeromonas spp.) may
grow as red colonies. Hence, confirmatory tests for E. coli
are not required.
Field study
Lysimeters The study was conducted in outdoor field
lysimeters. The PVC lysimeters (0.45 m I.D., 1 m tall)
were sealed at the bottom with PVC sheets (Fig.1). Sandy
soil was obtained from a field at the Macdonald Campus
of McGill University and was compacted to a bulk density
of 1,350 kg m-3 leaving a 0.05 m void space from the top.
Sampling ports were drilled radially in the lysimeter walls
at depths of 0.15 m, 0.36 m and 0.57 m from the top of the
lysimeter. Each sampling level had 4 sampling ports,
drilled radially, from which soil samples were collected to
make a composite sample for analysis. A 0.05 m diameter
perforated PVC pipe was installed at the bottom of each
lysimeter to allow leachate flow. The lysimeters were kept
outdoors in natural conditions. No crop was planted to
avoid uptake of water by plants; hand weeding was done
to remove weeds in the lysimeters. Meteorological data
were collected from the Ste-Anne-de-Bellevue station
(QC, Canada) of Environment Canada and the evaporation
rate was calculated by installing evaporation pans placed
at the lysimeter site. During the study period, the average
air temperature, humidity and evaporation rate were
19.1oC, 71.75%, and 2.1 mm/day, respectively.
Field experiment design A completely randomized
design with three replicate lysimeters of two different
biochar amendments (either SPB or FPB) and nonamended control was carried out in nine lysimeters. In the
biochar-amended treatments (soil+SPB and soil+FPB), the
top 0.10 m of this soil was amended with either SBP or
FPB in a proportion of 99:1 soil:biochar (214.6 g biochar
per lysimeter), which is the generally used ratio for
agricultural applications (DeLuca et al. 2006; Park et al.
2011). Raw bedding-free poultry manure was obtained
from the Donald McQueen Shaver Poultry Complex at the
Macdonald Campus of McGill University (Montreal, QC,
Canada). Immediately after the biochar was mixed into the
soil, 10.1 Mg ha-1 (161 g of manure per lysimeter) of
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Fig. 1. Schematic of the outdoor field PVC lysimeters.
poultry manure was applied on the lysimeter soil surface.
The lysimeters were irrigated by simulating the highest
total rainfall (173.4 mm) during the month of May in the
fifty-year period from 1962 to 2011; the amount was
calculated for May as a worst-case scenario (van Bochove
et al. 2010). Three irrigations (57.8 mm each) were applied
in 30 days, scheduled on days 0, 15, 30, and the same
amount in the fourth irrigation was applied 60 days after
the application of biochar and manure. All lysimeters were
placed under a canopy in order to prevent natural rainfall
and to apply simulated rainfall only.
Microbiological analysis of soil Prior to any biochar or
poultry manure application, and on days 2, 7, 14, 17, 21,
29, 32, 37, 59 and 62 after the application, composite
(from 4 ports at each depth) soil samples were drawn at
each of the three depths in each lysimeter and placed in
sterile Whirl-Pak bags (NASCO Corp., Fort Atkinson, WI,
USA) (3 depths = 3 composite soil samples from each
lysimeter). Care was taken to ensure all the sampling
materials were sterilized to prevent contamination.
Composite soil samples were transported to the laboratory
where they were analyzed for E. coli and total coliforms
on the same day. For each composite soil sample, 10 g of
soil were suspended in 90 mL of autoclaved saline
solution, and the diluted sample (10-1 dilution) was shaken
for 1 h on a rotary shaker operating at 250 rpm at room
temperature. Three replicates of 1 mL aliquots of the
diluted samples (10-1 dilution) were each added to 9 mL of
saline solution, and further 10-fold dilutions were done in
the same manner (up to 10-7). Each dilution was replicated
three times and further inoculated. A 100-µL aliquot of
each dilution was pipetted onto a Petri plate containing
Eosin Methylene Blue agar (EMB agar, Fischer Scientific,
Canada) and spread evenly using a sterilized disposable L-
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shaped hockey stick spreader (Fischer Scientific, Canada).
The Petri dishes were inverted and incubated for 24 h at
35±2oC. The EMB agar is selective and differential for
Gram-negative enteric bacteria such as E. coli and total
coliforms. E. coli appears as large, blue-black colonies
with a green metallic sheen, whereas total coliforms
appear as colorless to amber colonies. Colonies were
counted under 10-15 × magnification. The total coliforms
and E. coli loads were expressed in Log10 CFU g-1. The
results are presented, for each depth, as an average of all 3
lysimeters for each treatment group.
Microbiological analysis of leachates On days 1, 16, 31
and 61 after application or poultry manure and biochar,
leachate samples were collected from the drain of each
lysimeter in autoclaved glass bottles. Care was taken to
ensure all the sampling materials were sterilized to prevent
contamination. Leachate samples were transported to the
laboratory where they were analyzed for E. coli and total
coliforms on the same day. For each leachate sample, a 1
mL aliquot was serially diluted and, E. coli and total
coliforms were enumerated by membrane filtration as
described hereinabove.
Statistical analysis
The laboratory and leachate data from field lysimeters
were analyzed using the temporal repeated measures
analysis of variance (SAS institute Inc. 2009) and the
time-wise effect of treatments was evaluated. Soil data
from field lysimeters were analyzed by spatial (depth) and
temporal (day of sampling) repeated measures analysis of
variance; time and depth-wise effect of treatments was
evaluated.
RESULTS AND DISCUSSION
Characteristics of soil and biochar
Sandy soil was obtained from a field at the Macdonald
Campus of McGill University (Montreal, QC, Canada).
Physical properties of the sandy soil used in laboratory
experiments and used to fill the lysimeters are shown in
Table 1. The soil in the lysimeter was packed to a bulk
density equivalent to that observed in the field. The soil
had high sand content and thus, had relatively high
hydraulic conductivity. The soil was free from fecal
coliform (E. coli) but contained about 104 CFU of total
coliforms per g (Table 2). Two types of biochar procured
from BlueLeaf Inc. (Drummondville, QC, Canada) were
selected: slow pyrolysis biochar (SPB) and fast pyrolysis
biochar (FPB). SPB was produced from soft wood at
450°C for a duration of 2.5 hours; the pH was 9.35 and
moisture content was 8.9%. FPB was produced from hard
wood at >700°C over a period of a few minutes; the pH
was 8.65 and moisture content was 39.8%.
Fig. 2. Scanning electron microscope images of slow
pyrolysis biochar (SPB, A and B) and fast
pyrolysis biochar (FPB, C and D).
The particle size distribution of the two types of
biochar was determined by sieving (Table 3). The mineral
composition of the biochars was determined by proximate
analysis whereas hydraulic conductivity was determined
by the constant head method using 0.01 M CaCl2 solution
(Table 4). The saturated hydraulic conductivity of SPB and
FPB is relatively higher than that of the sandy soil. The
hydraulic conductivity is affected by the ash content of
biochar (Das et al. 2010); both biochars have relatively
low ash content, and thus, have reasonably high hydraulic
conductivity. The biochars were further characterized by
scanning electron microscopy (Hitachi S-3000N Variable
Pressure-SEM) (Fig. 2). It was observed that SPB has a
more distinct pore (≈200 µm mean diameter) structure
(Figs. 2A and 2B) as compared to FPB (Figs. 2C and 2D).
SPB also has a larger fraction of larger particles (Table 3).
Thus, SPB has a greater macropore space and surface area
than the FPB. From SEM imagery, it is evident that FPB
has a complex and fibrous structure with some void spaces
between fibres (Figs. 2C and 2D). The macropores appear
to be disintegrated but still they exist, especially those
visible on the top portion of the image (Figs. 2C and 2D).
The material looks very rugged.
Laboratory study
The E. coli loads in the solution (i.e., the non-adsorbed E.
coli) in the soil treatments are given in Table 5. It is clear
that the E. coli loads in the solution of the soil+SPB and
Table 1. Physical characteristics of the soil used in the laboratory studies and in the field study.
Soil type
Sandy
Sand
92.2%
Silt
4.3%
pH
5.5
* Standard deviation
1.24
Bulk density
1,350 kg m-3
Organic matter
Cation exchange capacity
Hydraulic conductivity
2.97%
4.9 cmol kg-1
1.0±0.3* mm min-1
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Table 2. Total coliforms and E. coli (fecal coliform)
loads in poultry manure and lysimeter soil at
different depths prior to manure application.
Bacterial load (CFU g-1)
Material
Poultry
manure
Soil
Soil
Soil
Fig. 3. Temporal evolution of the concentration of total
coliforms at A) 0.10 m, B) 0.31 m and C) 0.52 m
from the surface of the poultry manureamended soil in the outdoor field PVC
lysimeters. Error bars represent mean ±
standard deviation. Different letters denote the
concentrations significantly different (H0:
P<0.05) at the measurement depth and time.
Irrigations were applied to the lysimeters on
days 0, 15, 30 and 60.
soil+FPB treatments were significantly lower (about 100
times) than the E. coli load in the soil treatment after 24 h,
3 days and 7 days (H0: P<0.05). In the control treatment,
the slight increase in concentration after 3 days, as
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Depth (m)
Not
applicable
0.10
0.31
0.52
Total coliforms
1.50±0.33x10
8
E. coli
1.7±0.17x105
2.60±1.63x104
<1.0
1.43±0.91x10
4
<1.0
0.35±0.07x10
4
<1.0
compared to 24 h, is probably due to the release of E. coli
from soil. A marginal release of E. coli in soil+FPB is also
evident after 7 days. No such release is observed in
soil+SPB. It appears that the E. coli adsorption potential of
soil+SPB is slightly better than that of soil+FPB. This can
be attributed to the higher abundance of macropores in
SPB, which act as sites for adsorption, as seen in the SEM
images (Fig. 2). The particle size of SPB is larger as
compared to FPB and soil (Tables 3); thus, the holding
potential of the SPB pores is higher due to a larger surface
area of macropores in the latter type of biochar. For
practical purposes, it may be considered that SPB has
higher adsorption potential as compared to both soil and
FPB.
Field study
Total coliforms in soil The concentrations of total
coliforms in the lysimeters at three depths under all three
treatments monitored over the study period are shown in
Fig. 3. The concentration of total coliforms at three depths
in all treatments increased after the application of poultry
manure (Table 2 versus Fig. 3). At 0.1 m depth (Fig. 3A),
the concentration of total coliforms gradually increased in
all treatments until day 29. This could be due to the
migration of total coliforms from the manure applied on
the surface layer to the lower depths in the lysimeters after
irrigation. Although it was not apparent in the soil+SPB
and soil+FPB treatments, there was a decrease in
concentration of total coliforms in the soil treatment on
day 17 after irrigation was applied on day 15 (H0:
P<0.05), and it bounced back on day 21 (H0: P<0.05).
Crane et al. (1980) also observed slight re-growth after
initial decline. It is likely that greater loads of coliforms
adsorbed onto biochar in the soil+SPB and soil+FPB
treatments and therefore, there was minimal impact after
the first irrigation. A decrease in the concentrations of total
coliforms in all three treatments was observed on day 32
(H0: P<0.05), likely due to the effect of irrigation applied
on day 30. Then, the concentrations gradually decreased in
all three treatments (especially in the soil+SPB, H0:
P<0.05) and reached levels lower than the initial
concentrations observed on day 2. It appears that irrigation
water has some impact on the transport of total coliforms.
This could be due to dilution and leaching effects. The
results clearly indicated that the concentrations of total
coliforms in soil+SPB were lower than both the soil and
soil+FPB treatments throughout the study period (Fig.
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Table 3. Particle size distribution of slow pyrolysis
biochar (SPB) and fast pyrolysis biochar
(FPB).
Mesh size
(inch or #)
0.750”
0.625”
0.375”
0.250”
#10
#20
#40
#60
#80
#100
Pan
Total
Particle
diameter
(mm)
>19
16 to 19
9.5 to 16
6.3 to 9.5
2.0 to 6.3
0.85 to 2.0
0.425 to 0.85
0.25 to 0.425
0.18 to 0.250
0.15 to 0.18
<0.15
Biochar retained (%)
SPB
FPB
0.0
13.2
6.9
18.4
13.9
22.0
9.4
3.9
4.9
4.5
2.9
0.0
0.0
0.0
0.0
0.0
2.4
11.4
24.3
21.5
21.6
18.8
100.0
100.0
3A), as was observed in the laboratory studies (Table 5).
The statistical analysis revealed that at 0.1 m depth, the
concentrations of total coliforms in soil+SPB were
significantly lower than the concentrations measured in
both soil and soil+FPB (H0: P<0.05; Fig. 3A and Table 6).
The concentrations in the soil treatment were either higher,
or at par, with those of the soil+FPB (H0: P<0.05; Fig.
3A). The physical structure, greater surface area and
greater macro pore space of SPB might have caused
greater adsorption/retention of microbes in the upper layer
and/or decimated total coliforms.
At 0.31 m depth (Fig. 3B), the concentrations of total
coliforms increased in all of the treatments until 14 days:
this was likely due to the migration of total coliforms from
the upper layer after the application of poultry manure. A
sharp decrease in the concentrations was observed at this
depth on day 17 (H0: P<0.05) (Fig. 3B, after the irrigation
on day 15) as was observed in the soil treatment at 0.1 m
(Fig. 3A). This suggests that the concentration of total
coliforms may decrease temporarily after irrigation.
Fluctuations in the concentrations of total coliforms were
observed between days 17 and 37 in all three treatments at
0.31 m (Fig. 3B), whereas the concentrations dropped at
59 and 62 days (especially in the soil+SPB) and reached
levels lower than the initial concentrations observed on
day 2. Generally, the concentrations in soil+SPB were the
lowest whereas they were the highest in the soil treatment.
The statistical analysis showed that the concentrations of
total coliforms in soil+SPB were lowest, or at par, with the
other treatments.
Table 4. Mineral composition of slow pyrolysis biochar
(SPB) and fast pyrolysis biochar (FPB).
Constituent
Carbon (%)
Hydrogen (%)
Nitrogen (%)
Sulfur (%)
Oxygen (%)
Ash (total %)
Hydraulic conductivity (mm min-1)
SPB
FPB
77.00
2.20
0.56
0.02
3.00
17.30
1.60
74.00
3.20
0.42
0.02
12.60
9.80
2.60
At 0.52 m depth (Fig. 3C), the concentrations of total
coliforms followed cyclical patterns: the concentrations
gradually increased and then suddenly decreased at day 17
(after the day 15 irrigation) and day 32 (H0: P<0.05) (after
the day 30 irrigation). Again, the concentrations dropped at
59 and 62 days (especially in the soil+SPB) and reached
levels lower than the initial concentrations observed on
day 2 after the application of manure.
Figure 3 shows that between 0 and 37 days, the
concentration of total coliforms increased at 0.10 m,
marginally changed at 0.31 m, and slightly decreased at
0.52 m due to transport from surface soil. However, once
the mobility of total coliforms decreased or ceased, there
was a steep decline (between 57% at 0.10 m and 76% at
0.52 m) in their concentration from day 37 to day 62. A
laboratory study on the survival of total coliforms in dog
feces-amended soils at room temperature showed that at
lower moisture content, the half-life of total coliforms was
16.7 days, and 0.03% of total coliforms could survive 170
days (Adhikari et al. 2007): at a die-off rate of 0.041 day -1,
about 91% of total coliforms would die in 25 days in the
lysimeters used in the current study (Fig. 3). In the same
dog feces-amended soils but at a higher moisture content,
the half-life of total coliforms was 20.1 days and 0.09%
survival was observed after 170 days (Adhikari et al.
2007): at a die-off rate of 0.035 day-1, 87% of total
coliforms would die in 25 days in our lysimeters (Fig. 3).
Garcia-Orenes et al. (2007) conducted a survival study of
total coliforms in pots filled with three types of soils
amended with municipal sewage sludge in a greenhouse:
total coliforms disappeared in 33 to 40 days in the absence
of irrigation, but their survival extended to 85 to 90 days
when irrigation was applied. It is known that substrate
availability for soil microorganisms increases with
increased organic matter content of the soil, which in turn
extends the survival of coliforms (Klein and Casida 1967;
Mallmann and Litsky 1951). In the current study, sandy
soil with a relatively high organic matter content was used
Table 5. E. coli (fecal coliform) loads* in solution with slow pyrolysis biochar (SPB) and fast pyrolysis biochar (FPB)
treatments in laboratory study.
Treatment
24 h
3 days
a
a
Soil
1.50±0.13 x106
1.70±0.11 x106
b
4
b
Soil:SPB
<1.00 x10
<1.00 x104
b
b
Soil:FPB
3.00±2.00 x104
<1.00 x104
-1
*: E. coli loads (mean±standard deviation) are CFU mL .
a, b
: Values with different superscript letters are significantly different for the sampling day (H0: P<0.05).
1.26
LEGÉNIEDESBIOSYSTÈMESAUCANADA
7 days
a
8.70±0.80 x105
b
<1.00 x103
b
1.17±0.15 x104
AriefIsmailetal.
3
4
Table 6. Spatio-temporal repeated measures analysis of
total coliforms and E.coli (fecal coliform) loads
at three depths over ten measurement dates
after application of poultry manure.
Factor
Treatment
Depth
Time
Treatment x Depth
Treatment x Time
Depth x Time
Treatment x Depth x Time
Fig. 4. Temporal evolution of the concentration of total
coliforms in leachates from the outdoor field
PVC lysimeters. Error bars represent mean ±
standard deviation. Different letters with the
bars denote the concentrations significantly
different (H0: P<0.05). Irrigations were applied
on days 0, 15, 30 and 60.
(Table 1) and irrigation was applied, both of which would
have contributed to the presence of total coliforms in
lysimeter soil at all three depths after 62 days (Fig. 3).
The differences in concentrations and different rates
of change with treatments and depth over time reflected
the significant effect of treatment, depth, time and their
interactions (Table 6). Overall, SPB amendment in the soil
surface layer was the most effective in reducing the
concentrations of total coliforms in soil as compared to
non-amended soil as well as soil+FPB.
Total coliforms in leachate Total coliforms were detected
in the leachates collected from the three treatments during
all four irrigation events (Fig. 4). This indicates that total
coliforms are quite mobile (vertically) in sandy soil and
they could migrate at least 1 meter deep. It was previously
reported that the concentration of coliforms (Entry et al.
2000) and enteric bacteria (Paluszak et al. 2003) decreased
with depth of soil. In the present study, the concentrations
of total coliforms in soil gradually decreased with depth
down to 0.52 m (Fig. 3), and thus, it is possible that the
concentration in the bottom part of the lysimeters would be
quite low. However, the presence of total coliforms in the
lysimeter leachates suggests that they can migrate deeper
in the soil profile and reach the leachate. The transport of
microbes through porous medium by flow of water is
called passive transport. In such a case, microbial motility
itself does not play a significant role in movement, which
is primarily tied to flow (Robert and Chenu 1992). Smith
et al. (1983) reported that a high concentration of
coliforms could move downwards through 0.28 m in soil
columns whereas Stoddard et al. (1998) observed that total
coliforms could migrate vertically more than 0.90 m
Volume58
2016
Total
coliforms
*
*
*
*
*
*
*
E. coli
*
*
*
*
*
*
*
through soil. Smith et al. (1983) and Abu-Ashour et al.
(1998) observed that bacterial transport through soil was
greatly increased by the presence of macropores. Finally,
Unc and Goss (2003) corroborated that bacterial transport
was mainly due to water flowing through macropores.
In all three treatments, the concentration of total
coliforms in the leachates from the second irrigation (Fig.
4, day 16) was higher as compared to the first irrigation
(day 1) probably due to the cumulative leaching effect of
these first two irrigations. It can be seen from Fig. 3C that
there was a decrease in the concentration of total coliforms
at 0.52 m, especially in the soil+SPB treatment, after the
second irrigation at day 30. Thus, it is likely that total
coliforms were transported with water and detected in the
leachate. The concentrations of total coliforms in the
leachate were also high after the third irrigation (day 31),
although slightly lower as compared to day 16. In the later
stage of the experiment, the concentration of total
coliforms decreased both in soil at all three depths (Fig. 3)
and in the leachate (Fig. 4, day 61) (H0: P<0.05) perhaps
due to the natural die-off of these types of bacteria.
In the first three irrigations, the concentrations of total
coliforms in the control were significantly higher (H0:
P<0.05) as compared to the biochar treatments (Fig. 3).
The concentrations in soil+SPB were significantly lower
than, or at par, with the soil+FPB. The concentrations in
the leachates from the fourth irrigation (day 61) were low
and statistically not different between all treatments,
although numerically the concentration was the highest in
the control and the lowest in FPB. Thus, the concentration
of total coliforms was generally the lowest in the soil+SPB
treatment, whereas it was always the highest in the control
(soil alone). These results are consistent with the
observation that the concentration of bacteria in drainage
water is proportional to the bacterial concentration in soil
(Evans and Owens 1972). The results indicate that biochar
amendment in the surface layer, especially SPB, was
effective in mitigating the transport of total coliforms not
only in the soil but also in the leachates. Therefore, SPB
amendment would be an cost-effective method for
controlling total coliforms threats to surface and
subsurface water bodies.
E. coli in soil Analysis of lysimeter soil samples prior to
the poultry manure amendment showed that E. coli was
CANADIANBIOSYSTEMSENGINEERING
1.27
5
6
Fig. 6. Temporal evolution of the concentration of E.
coli in leachates from field lysimeters. Error
bars represent mean ± standard deviation.
Different letters with the bars denote the
concentrations significantly different (H0:
P<0.05). Irrigations were applied on days 0, 15,
30 and 60.
Fig. 5. Temporal evolution of the concentration of E.
coli at: A) 0.10 m, B) 0.31 m and C) 0.52 m from
the surface of the poultry manure-amended soil
in outdoor field PVC lysimeters. Error bars
represent mean ± standard deviation. Different
letters denote the concentrations significantly
different (H0: P<0.05). Irrigations were applied
on days 0, 15, 30 and 60.
not detected in the soil at all three depths whereas a high
concentration of E. coli was found in poultry manure
(Table 2). Therefore, any E. coli in the lysimeter soil
would originate from the poultry manure. The
concentrations of E. coli in the lysimeter soil over the
study period are shown in Fig. 5. Within 2 days of the
application of poultry manure, followed by irrigation, the
concentrations of E. coli in the soil at 0.1 m dramatically
1.28
increased (Table 2 versus Fig. 5A) due to the massive
transport of E. coli from the poultry manure to the soil.
Thereafter, the concentrations of E. coli in the soil
increased between day 2 and day 37 (H0: P<0.05) and
decreased slightly until the end of the monitoring period,
which is a reflection of either much lower transport rates
of E. coli from manure to soil or the depletion of E. coli
populations in the manure during these samplings. At 0.31
m (Fig. 5B) and 0.52 m (Fig. 5C) depths, it is noteworthy
that (i) the concentrations of E. coli in un-amended soil
followed the same pattern as the one observed at 0.10 m,
whereas (ii) increases in the concentrations of E. coli in
soil+SPB and soil+FPB were observed only after 14 days
at 0.31 m and after 29-32 days at 0.52 m (H0: P<0.05).
This shows that SPB and FBP amendments on the top soil
layer of the lysimeters played a significant role in retarding
the downward migration of E. coli. At all three depths
(Fig. 5A, 5B and 5C), (iii) the concentrations of E. coli
were consistently and significantly higher (H0: P<0.05) in
the control (un-amended soil) as compared to both biochar
treatments, except for the soil+FPB treatment at 0.10 m
between days 37 and 62; (iv) the concentrations of E. coli
in soil+SPB were either significantly lower than, or at par
with, those of soil+FPB (H0: P<0.05). This shows that
SPB is the most effective in reducing the migration of E.
coli in the soil matrix. However, in contrast to what was
observed with total coliforms (Fig. 3), the concentrations
of E. coli were maintained throughout the 62-day
monitoring period at all three depths and in all three
LEGÉNIEDESBIOSYSTÈMESAUCANADA
AriefIsmailetal.
7
8
treatments (Fig. 5). This demonstrates that E. coli may
remain viable for extended periods (few to several months)
once it has entered the soil-water system.
A laboratory study conducted by Crane et al. (1980)
showed that the die-off rate of fecal coliform varied from
0.032 to 0.342/day: from this data, it can be extrapolated
that the die-off of fecal coliform in these conditions in 60
days would vary between 98.6% and 100%. Zhai et al.
(1995) calculated die-off rates of 0.04/day to 0.88/day
depending on the type of soil and the incubation time; at
this rate, fecal coliform should die-off in 60 days. Estrada
et al. (2004) observed that the concentration of faecal
coliforms, and E. coli decreased considerably or was not
detected in soil/sludge mixtures after 80 days in both
outdoor and laboratory conditions. However, a
considerable increase in the concentration of
microorganisms was observed when calcium ammonium
nitrate was applied as a nutrient to the soil/sludge
mixtures. E. coli may die-off when soil conditions are
inadequate but can establish as a member of the soil
microbiota or even grow in soil if nutrients and moisture
are available at a suitable temperature (Byappanahalli and
Fujioka 1998; Gagliardi and Karns 2000). Given the soil in
the lysimeters was brought from an agricultural field, it
contained nutrients. Further more irrigation had been
applied to the soil; therefore, E. coli concentration would
have increased in the lysimeters.
The difference in concentrations and different rates of
change with treatments and depth over time reflected the
significant effect of treatment, depth, time and their
interactions (Table 6). Overall, SPB amendment on the
soil surface was the most effective in reducing the
concentrations of E. coli in soil as compared to nonamended soil as well as soil+FPB.
E. coli in leachate E. coli was present in the leachates
collected from the control lysimeters (soil) during all four
irrigation events, but concentrations were below detection
limit in the leachates from biochar-amended lysimeters
(soil+SPB and soil+FPB, Fig. 6). This is rather unexpected
since the concentrations of E. coli were maintained
throughout the entire monitoring period at all three depths
and in all three treatments (Fig. 5). However, since the
difference in the concentrations of E. coli between the
control lysimeters (soil) and the biochar-amended
lysimeters (soil+SPB and soil+FPB) increased with depth
(Fig. 5), it may be possible that E. coli did not migrate
down to the bottom of the soil+SPB and soil+FPB
lysimeters (i.e., 1 m) where the leachates were collected.
In accordance with what was observed in Fig. 5, the
concentrations of E. coli were stable throughout the 61-day
monitoring period in the leachate of the control lysimeters
(Fig. 6, Soil). Again, this demonstrates that E. coli may
remain viable for extended periods (few to several months)
once it has entered the soil-water system. For example, E.
coli concentrations of 101 to 105 per litre of drainage water
were observed during a 4-month period after animals had
stopped grazing in a pasture (Evans and Owens 1972).
Volume58
2016
CONCLUSIONS
In a laboratory study, higher adsorption of E. coli was
observed with soil+biochar, especially soil+SPB, than
sandy soil alone. In field lysimeters, with irrigation, E. coli
moved from the topsoil to depths of more than 0.5 m in the
soil treatment; however, minimal E. coli concentrations
were observed at depths below 0.1 m in the biocharamended soils. Although the concentration gradually
increased at all the depths in all treatments with time, the
concentrations were lowest in soil where the SPB was
added. There was minimal concentration of E. coli in the
leachate from both biochar treatments. This showed that
biochars, especially SPB, are suitable for reducing E. coli
concentrations in drainage waters. The results of the
lysimeter study also demonstrated that when poultry
manure was applied, total coliforms could travel to a
greater depth than 0.5 m, whether or not biochars were
applied at the surface. However, the coliform
concentrations were the lowest in the soil treated with
SPB. The concentration decreased with depth and time.
This showed that SPB was more effective in filtering total
coliform, moving deep into the soil, and flowing out in the
leachate. Thus, it appears that biochar amendment at the
surface can mitigate transport of manure-borne pathogens
in drainage water.
ACKNOWLEDGEMENTS
This work was supported by a National Sciences and
Engineering Research Council of Canada (NSERC)
research grant. Additional financial support was provided
by IC-IMPACTS (the India-Canada Centre for Innovative
Multidisciplinary Partnerships to Accelerate Community
Transformation and Sustainability).
REFERENCES
Abu-Ashour, J., D.M. Joy, H. Lee, H.R. Whiteley, and S.
Zelin. 1998. Movement of bacteria in unsaturated
soil columns with macropores. Transactions of the
ASAE 41(4): 1043-1050.
http://dx.doi.org/10.13031/2013.17267
American Society of Agricultural Engineers. 2005.
Manure Production and Characteristics. ASAE
Standard D384.2 MAR2005. St. Joseph, MI: ASAE.
Accessed 21 March 2013 at http://evo31.
ae.iastate.edu/ifafs/doc/pdf/ASAE_D384.2.pdf.
Bird, M. I., Moyo, C., Veenendaal, E. M., Lloyd, J., Frost,
P., 1999. Stability of elemental carbon in a savanna
soil. Global Biogeochemical Cycles 13: 923-932.
Atidégla, S.C., J. Huat, E.K. Agbossou, H. Saint-Macary,
and R.Gl. Kakai. 2016. Vegetable Contamination by
the Fecal Bacteria of Poultry Manure: Case Study of
Gardening Sites in Southern Benin. International
Journal of Food Science 2016: Article ID 4767453:
1-8. http://dx.doi.org/10.1155/2016/4767453
Bolton, D.J., C.M. Byrne, J. Sheridan, D.A. McDowell,
I.S. Blair, and T. Hegarty. 1999. The survival
characteristics of a non-toxigenic strain of
Escherichia coli O157:H7. Journal of Applied
Microbiology 86(3): 407-411.
http://dx.doi.org/10.1046/j.1365-2672.1999.00677.
CANADIANBIOSYSTEMSENGINEERING
1.29
0
9
Borst, M., and A. Selvakumar. 2003. Particle-associated
microorganisms in stormwater runoff. Water
Research 37: 215–223.
http://dx.doi.org/10.1016/S0043-1354(02)00244-0
Chevalier, P., P. Levallois and P. Michel. 2004.
Waterborne enteric infections potentially linked to
livestock production: a literature review. Vecteur
Environnement 37: 90-106.
Crane, S.R., P.W. Westerman, and M.R. Overcash. 1980.
Die-off of fecal indicator organisms following land
application of poultry manure. Journal of
Environmental Quality 9:531-537.
http://dx.doi.org/10.2134/jeq1980.004724250009000
30042x
Dalahmeh, S.S., M. Pell, B. Vinneras, L.D. Hylander, I.
Oborn, and H. Jonsson. 2012. Efficiency of bark,
activated charcoal, foam and sand filters in reducing
pollutants from greywater. Water Air and Soil
Pollution 223: 3657-3671.
http://dx.doi.org/10.1007/s11270-012-1139-z
Das, K.C., K. Singh, B.P. Bibens, R. Hilten, S.A. Baker,
W.D. Greene, and J.D. Peterson. 2010. Pyrolysis
characteristics of forest residues obtained from
different harvesting methods. Applied Engineering
in Agriculture 27(1): 107-113.
http://dx.doi.org/10.13031/2013.36215
Diarra, M.S. and F. Malouin. 2014. Antibiotics in
Canadian poultry production and anticipated
alternatives. Frontiers in Microbiology 5, article 2:115. http://dx.doi.org/10.3389/fmicb.2014.00282
Downie, A., A. Crosky, and P. Munroe. 2009. Chapter 2.
Physical properties of biochar. In: Biochar for
Environmental
Management
Science
and
Technology, 13-32, J. Lehmann and S. Joseph, eds.
London: Earthscan Publishers Ltd.
Entry, J.A., R.K. Hubbard, J.E. Thies, and J.J. Fuhrmann.
2000. The Influence of vegetation in riparian
filterstrips on coliform bacteria: II. Survival in soils.
Journal of Environmental Quality 29(4): 1215-1224.
http://dx.doi.org/10.2134/jeq2000.004724250029000
40027x
Estrada, I.B., A. Aller, F. Aller, X. Gomez and A. Moran.
2004. The survival of Escherichia coli, faecal
coliforms and enterobacteriaceae in general in soil
treated with sludge from wastewater treatment plants.
Bioresource Technology 93: 191-198.
Evans, M.R. and J.D. Owens. 1972. Factors affecting the
concentration of faecal bacteria in land-drainage
water. Journal of General Microbiology 71: 477-485.
Furtula, V., E.G. Farrell, F. Diarrassouba, H. Rempel, J.
Pritchard, and M.S. Diarra. 2010. Veterinary
pharmaceuticals and antibiotic resistance of
Escherichia coli isolates in poultry litter from
commercial farms and controlled feeding trials.
Poultry Science 89: 180–188.
http://dx.doi.org/10.3382/ps.2009-00198
1.30
Health Canada. 2006. Guidelines for Canadian Drinking
Water Quality: Guideline Technical Document —
Total Coliforms. Ottawa, Ontario: Water Quality and
Health Bureau, Healthy Environments and
Consumer Safety Branch, Health Canada. Accessed
21 March 2013 at http://www.hc-sc.gc.ca/ewhsemt/alt_formats/hecs-sesc/pdf/pubs/watereau/coliforms-coliformes/coliforms-coliformeseng.pdf.
Health Canada. 2008. Environmental and workplace
health. Accessed 21 March 2013 at http://www.hcsc.gc.ca/ewh-semt/pubs/water-eau/well-puitseng.php.
Health Canada. 2011. Escherichia coli in Drinking Water.
Document for Public Comment. Ottawa, ON: The
Federal-Provincial-Territorial
Committee
on
Drinking Water, and Health Canada. Accessed
March 21 2013 at http://www.hc-sc.gc.ca/ewhsemt/alt_formats/pdf/consult/_2011/ecoli/ecoli-draftebauche-eng.pdf.
International biochar initiatives. 2016. Biochar use in
soils.
Accessed
23
January
2016
at
http://www.biochar-international.org/biochar/soils.
Jamieson, R.C., R.J. Gordon, K.E. Sharples, G.W.
Stratton, and A. Madani. 2002. Movement and
persistence of fecal bacteria in agricultural soils and
subsurface drainage water: A review. CBE 44:1.11.9.
Jenkins, M.B.,D.M., Endale, H.H. Schomberg, and R.R.
Sharpe. 2006. Fecal bacteria and sex hormones in
soil and runoff from cropped watersheds amended
with poultry litter. Science of Total Environment
358 (1-3): 164-177.
http://dx.doi.org/10.1016/j.scitotenv.2005.04.015
Lal, R. (ed.). 2006. Encyclopedia of Soil Science, 2nd
edition. 2 vols. Boca Raton, FL: CRC Press.
http://dx.doi.org/10.1017/S0014479706214108
Lehmann, J. M.C. Rillig, J. Thies, C.A. Masiello, W.C.
Hockaday, and D. Crowley. 2011. Biochar effects on
soil biota – A review. Soil Biology and Biochemistry
42: 1812-1836.
http://dx.doi.org/10.1016/j.soilbio.2011.04.022
Liang, B., J. Lehmann, D. Solomon, J. Kinyangi, J.
Grossman, B. O'Neill, J.O. Skjemstad, J. Thies, F.J.
Luizao, J. Petersen, and E.G. Neves. 2006. Black
carbon increases cation exchange capacity in soils.
Soil Science Society of America Journal 70(5):
1719-1730.
http://dx.doi.org/10.2136/sssaj2005.0383
McDowell-Boyer, L.M., J.R. Hunt, and N. Sitar. 1986.
Particle transport through porous media. Water
Resources Research 22(13): 1901-1921. doi:
10.1029/WR022i013p01901
http://dx.doi.org/10.1029/WR022i013p01901
Megchún-García, J.V, C. Landeros-Sánchez, A. SotoEstrada, M.d.R. Casta-eda-Chávez, J.P. MartínezDávila, I. Nikolskii-Gavrilov, I. Galaviz-Villa, and
F. Lango-Reynoso. 2015. Total Coliforms and
LEGÉNIEDESBIOSYSTÈMESAUCANADA
AriefIsmailetal.
1
2
Escherichia coli in Surface and Subsurface Water
from a Sugarcane Agroecosystem in Veracruz,
Mexico. Journal of Agricultural Science 7(6): 110119.
http://dx.doi.org/10.5539/jas.v7n6p110
Moges, M.E., F.E. Eregno, A. Heistad. 2015. Performance
of biochar and filtralite as polishing step for on-site
greywater treatment plant. Management Of
Environmental Quality: An International Journal
26(4): 607-625.
http://dx.doi.org/10.1108/MEQ-07-2014-0101
Nicholson, F.A., S.J. Groves, M.L. Hutchison, N.
Nicholson, and B.J. Chambers. 2002. Pathogens in
animal manures: their survival during storage and
following land application. In: 10th International
Conference of the RAMIRAN Network (Štrbské
Pleso, Slovakia, May 14-18, 2002). 41-44, J.
Venglovský and G. Gréserová, eds. Košice, Slovak
Republic: University of Veterinary Medicine.
Consulted
21.03.2013
at
www.ramiran.net/DOC/A4.pdf.
Ogawa, M., Y. Yamabe, and G. Sugiura. 1983. Effects of
charcoal on the root nodule formation and VA
mycorrhiza formation of soy bean. The Third
International Mycological Congress (IMC3),
Abstract 578.
Omarova, M.N., L.Z. Orakbay, I.H. Shuratov, A.T.
Kenjebayeva, A.B. Zhumagalieva, and A.B.
Sarsenov. 2016. Ranking of the ecological disaster
areas according to coliform contamination and the
incidence of acute enteric infections of the
population in Kyzylorda region. International
Journal of Environmental and Science Education
11(11): 4093-4103.
Paluszak, Z., A. Ligocka, B. Breza-Boruta. 2003.
Effectiveness of Sewage Treatment Based on
Selected Faecal Bacteria Elimination in Municipal
Wastewater Treatment Plant in Toruń. Polish Journal
of Environmental Studies 12(3):345-349.
Pappas, E.A., R.S. Kanwar, J.L. Baker, J.C. Lorimor, and
S. Mickelson. 2008. Fecal indicator bacteria in
subsurface drain water following swine manure
application. Transactions of the ASABE 51(5):
1567-1573.
http://dx.doi.org/10.13031/2013.25313
Pietikäinen, J., O. Kiikkilä, and H. Fritze. 2000. Charcoal
as a habitat for microbes and its effect on the
microbial community of the underlying humus.
Oikos 89(2): 231–242. doi: 10.1034/j.16000706.2000.890203.
Robert, M., C. Chenu. 1992. Interactions between soil
minerals and microorganisms. In: Soil Biochemistry
(Vol. 7). 307-404, G. Stotzky and J.M. Bollag, eds.
New York: Marcel Dekker.
Rogers S. and J. Haines. 2005. Detecting and mitigating
the environmental impact of fecal pathogens
originating from confined animal feeding operations:
Review. National Risk Management Laboratory.
Office of research and development. United States
Environmental Protection Agency. Cincinnati. OH
45268
Volume58
2016
Rose, J.B., R.M. Atlas, C.P. Gerba, M.J. Gilchrist, M.W.
LeChevallier, M.D. Sobsey, and M.V. Yates. 1999.
Microbial Pollutants in Our Nation's Water:
Environmental
and
Public
Health
Issues.
Washington,
DC:
American
Society
for
Microbiology. Accessed 21 March 2013 at
http://www3.abe.iastate.edu/AE520/MicrobialPolluti
on.pdf
Saito, M. and T. Marumoto. 2002. Inoculation with
arbuscular mycorrhizal fungi: The status quo in
Japan and the future prospects. Plant and Soil 244(12): 273–279.
http://dx.doi.org/10.1023/A:1020287900415
Shanon, M.A., P.W. Bohn, M. Elimelech, J.G. Georgiadis,
B.J. Marinas, and A.M. Mayes. 2008. Science and
technology for water purification in the coming
decades. Nature 452: 301-310.
http://dx.doi.org/10.1038/nature06599
Smith, M.S., G.W. Thomas, R.E. White, and D. Ritonga.
1983. Transport of Escherichia coli through intact
and disturbed soil columns. Journal of
Environmental Quality 14 (1): 87-91.
http://dx.doi.org/10.2134/jeq1985.00472425001400
010017x
Stoddard, C.S., M.S. Coyne, and J.H. Grove. 1998. Fecal
bacteria survival and infiltration through a shallow
agricultural soil: Timing and tillage effects. Journal
of
Environmental
Quality
27:1516-1523.
http://dx.doi.org/10.2134/jeq1998.00472425002700
060031x
Stout, W.L., Y.A. Pachepsky, D.R. Shelton, A.M.
Sadeghi, L. Saporito, and A.N. Sharpley. 2005.
Runoff transport of faecal coliforms and phosphorus
released from manure in grass buffer conditions.
Letters in Applied Microbiology 41(3), 230–234.
http://dx.doi.org/10.1111/j.1472-765X.2005.01755.x
Thies, J.E., and M.C. Rillig. 2009. Characteristics of
Biochar: Biological Properties. In: Biochar for
Environmental
Management:
Science
and
Technology, 85-102, J. Lehmann and S. Joseph, eds.
London: Earthscan Publishers Ltd.
Unc, A. and M.J. Goss. 2003. Movement of fecal bacteria
through the vadose zone. Water Air and Soil
Pollution 149: 327–337.
http://dx.doi.org/10.1023/A:1025693109248
van Bochove, E., E. Topp, G. Thériault, G., J.T. Denault,
S. Allaire, F. Dechmi, and A.N. Rousseau. 2010.
Indicator of the Risk of Water Contamination by
Coliforms. In: Environmental Sustainability of
Canadian Agriculture: Agri-Environmental Indicator
Report Series - Report No. 3. Ottawa: Agriculture
and Agri-food Canada. posted 11 February 2010.
Accessed
21
March
2013
at
http://www4.agr.gc.ca/AAFC-AAC/displayafficher.do?id=1296059359221&lang=eng.
Warnock, D.D., J. Lehmann, T.W. Kuyper, and M.C.
Rillig. 2007. Mycorrhizal responses to biochar in
soil – concepts and mechanisms. Plant and Soil
300(1): 9–20.
http://dx.doi.org/10.1007/s11104-007-9391-5
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