IJERD – International Journal of Environmental and Rural Development (2010) 1–1

Research articleerd

Management of Manure Taking into Account of E.coli Lossfrom Farmland
YU SAITO
Graduate School of Agriculture, Tokyo University of Agriculture,
Tokyo, Japan
Email:saichu0136@yahoo.co.jp

MACHITO MIHARA
Faculty of Regional Environment Science, Tokyo University of Agriculture,
Tokyo, Japan
Email:m-mihara@nodai.ac.jp
Received 9 January 2010

Accepted 5 March 2010

Abstract Manure fermented insufficiently includes pathogen such as E.coli threatens theenvironment and human health. Thus, a proper management of the excrement is an
important issue. In addition, E.coli can be easily transported by surface runoff orpercolation from farmland where manure had been applied. So, the loss of E.coli fromfarmlands and its survival limit in manure are big concern from the viewpoints ofenvironmental conservation. Model experiments were conducted under artificial rainfall
simulator to investigate E.coli loss. Slope plots were filled with soil and then applied 3
types of manure; fresh cow dung, manures fermented for 14 days and 60 days. It wasproven that E.coli losses occurred even from the manure which was fermented for 60 days.
In addition, to acquire a proper treatment of manures, the experiments were conducted toinvestigate the survival limit of E.coli in the manure. It was observed that E.coli vanishedat the temperature higher than 60 degrees Celsius. Furthermore, the reduction of watercontent was not enough to eliminate E.coli. So, it became clear that controlling
temperature of manure is important for vanishing E.coli existing in manures.
Keywords E.coli, surface runoff, manure, temperature, water content

INTRODUCTION
Large amounts of livestock excrements are being produced in Japan. The Ministry of Agriculture,Forestry and Fisheries of Japan estimated 9,000,000 tons of livestock excrements are being
produced per year, and 60% of total livestock excrements are cow dung. At present, these large
amounts of excrements are being utilized as manure for improving soil condition. However, large
amounts of manure production lead to insufficient fermentation of cow dung. In developing
countries, it is difficult to ferment cow dung sufficiently, because of the lack of tools or equipments.
There were reports that insufficient fermented manure includes pathogens such as E.coli, amicroorganism which is normally present in intestinal tract of animals (Chun-Ming Gong et al.,2005). This bacterium will easily propagate in the surrounding environment and can be a threat tosanitation once being discharged with the excrements (Saito et al., 2006). Also, E.coli can be easilytransported by surface runoff along with soil sediments during heavy rainfall or by percolation intogroundwater (Yagura et al., 2006). Thus, a proper treatment of cow dung before applying tofarmlands became big concern. Therefore, research interests have been paid to the survival limits ofE.coli in manure for preventing the loss of E.coli from farmlands. So, the objective of this study
was to investigate the proper treatment of manure from the viewpoints of environmental
conservation.

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EXPERIMENTAL METHODS
Experiment on E.coli loss
Model experiments were conducted to investigate the E.coli loss from farmlands employing slope
plots under artificial rainfall simulator as shown in Fig. 1. Cow dung was utilized as the material
source of E.coli in this study, as it includes more E.coli than other animal excrements. The
properties of each material are shown in Table 1. The cell numbers of E.coli in fresh cow dung, 14days fermented manure and 60 days fermented manure were 4.1 x 107, 1.6 x 105 and 8.0 x 104 cfu/g
dry matters, respectively.
Three stainless slope model plots were prepared at 8 degrees as it is the maximum slope limiton handling agricultural machineries in sloping lands. Soil was compacted into the slope plots at
1.00 ± 0.06 g/cm3 in dry density (Kawai et al., 2007) and then cow dung or 2 kinds of manure wereapplied. Fresh cow dung was spread at Plot 1, while Plot 2 and Plot 3 were spread with fermented
manures for 14 days and 60 days, respectively. Both cow dung and fermented manures werebroadcasted into each plot uniformly. In order to observe and compare the losses of E.coli andorganic matter from 3 types of plot, artificial rainfall at 60mm/h intensity was simulated for 2 hours.
Discharge of surface runoff was measured at certain interval and sampled from each plots. TheE.coli in each sample was cultured by the spread plate method on the XM-G agar at 37 degrees
Celsius. After 20 hours, the number of blue spots showing E.coli colonies appeared on the agar wascounted. On the other hand, organic matter was analyzed by means of the ignition loss method.
Plot 1

Artificial rainfall simulator

Plot 2Plot 3

ManureSoil

Plot 1: Cow dung
Plot 2: Manure fermented for 14 days (Manure I)
Plot 3: Manure fermented for 60 days (Manure II)

Fig. 1 Slope model experiment

Table 1 Properties of materials
SampleCow dung
Manure IManure IISub-material
SawdustSawdustSawdustFermentation term

E.coli(day)(cfu/g dry matter)

01460

Water content
7

4.1 x 10
1.6 x 105
8.0 x 104

(%)
82
73
72

Experiment on survival limit
Another experiment was conducted to investigate the survival condition of E.coli in the manure. As
shown in Tables 2 and 3, cow dung and manure fermented for 14 days were used in thisexperiment. The change in E.coli cell number was observed under different temperature or watercontent.
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Cow dung or manure fermented for 14 days was enclosed with plastic wrap to avoid thereduction of water content and kept in the incubator under the different temperature at 30, 40, 50
and 60 degrees Celsius. Additionally, the changes in E.coli cell number with water content wereobserved. Cow dung or manure fermented for 14 days was kept in the greenhouse for enhancing
evaporation, and then certain amounts of samples were collected occasionally for analyzing E.coli
cell number as well as water content.
Table 2 Properties of cow dung or manure used in experiment at different temperature
SampleSub-material

Fermentation term

E.coli

Water content(day)(cfu/g dry)

(%)

782
72

Cow dung

Sawdust01.8 x 10

Manure I

Sawdust14

1.4 x 103

Table 3 Properties of cow dung or manure used in experiment at different water content
SampleSub-material

Fermentation term(day)

E.coli(cfu/g dry)

Water content(%)

Cow dung
Manure I

SawdustSawdust0
14

1.4 x 1088.6 x 105

7970

RESULTS AND DISCUSSION
E.coli loss from plots applied cow dung or manure
As shown in Fig. 2, E.coli loss from the plot applied fresh cow dung was the highest among allplots at 99% confidence interval. The loss of E.coli was observed not only from the plot applied
fresh cow dung but also from the plot applied manure fermented for 14 days or 60 days. Figs. 3, 4and 5 show the correlation between the amounts of E.coli and organic matter. It showed that E.coli
loss increased with organic matter in sediments.

84x10
4.0E+08abb

E.coli , cfu/LE.coli , cfu/m2

1.0E+121012

1.0E+061060
1.0E10+00R = 0.9735**
22x1082.0E+0800.0E+00
Cow dung

M anurefermented for
14 days

M anurefermented for
60 days0.00.5

1.0

1.5

2.0

2.5

Organic matter, g/L*There was a significant difference in total surface E.coli loss
at 99% between a and b.

Fig. 3 Relationship between E.coli andorganic matter in plot applied
cow dung

Fig. 2 E.coli loss by surface runoff during
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65.0E+05
5.0x105
E.coli , cfu/LE.coli , cfu/L

5.0x105.0E+06
2

R = 0.9006 **62.5x10
2.5E+06

2

R = 0.8011 **52.5x10
2.5E+05

00.0E+00

00.0E+00

0.0

0.5

1.0

0.0

1.5

0.4

0.8

1.2

Organic matter, g/L

Organic matter, g/L

Fig. 4 Relationship between E.coli andorganic matter in plot appliedmanure fermented for 14 days

Fig. 5 Relationship between E.coli andorganic matter in plot appliedmanure fermented for 60 days

Change in E.coli cell number
Based on the results of the experiment employing the incubator, the changes in E.coli cell number
at different temperature were summarized in Figs. 6 and 7. The cell number of E.coli started todecrease at 50 degrees Celsius in either cow dung or manure; however it did not vanish completely.
It was observed that E.coli vanished completely at 60 degrees Celsius in either cow dung or manure.
In opposition, there was no significant difference in E.coli cell number among various temperatures
under 50 degrees Celsius in either cow dung or manure.
The results of the experiment on the changes in E.coli cell number with water content weresummarized in Figs. 8 and 9. Water content decreased to 10% in either cow dung or manure for 9days. There was a tendency for E.coli cell number in either cow dung or manure to decrease withthe decrease in water content. Additionally, the fluctuation of E.coli cell number in cow dung wascomparatively smaller than that in manure. Although E.coli cell number at 28 days passed wassignificantly smaller than that at initial stage at 99% confidence interval, the reduction of watercontent was not enough for vanishing E.coli.

60℃

50℃

40℃

1.0E+04
10430℃
E.coli , cfu/g
dry matter

E.coli , cfu/gdry matter

1.0E+08108

1.0E+04
10460℃

50℃

40℃

30℃

1.0E+02102100
1.0E+00

1.0E+00
100
2H

4H

2H

6H

Fig. 6 Changes in E.coli with temperaturein cow dung

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4H

6H

Fig. 7 Changes in E.coli with temperature
in manure fermented for 14 days

IJERD – International Journal of Environmental and Rural Development (2010) 1–1E.coliaWater content
100bcccc105
1.0E+0550010
1.0E+00

Water content

E.coli , cfu/g
dry matter

1010
1.0E+100049152128day*The data followed by different letters differs significantly at 99% confidence interval.

Fig. 8 E.coli cell number with reduction of water content in cow
dung

E.coliE.coli , cfu/g
dry matter10
1.0E+06

Water content

ab80
bbbb103
1.0E+0340010
1.0E+00

Water content

6004

9152128

day*The data followed by different letters differs significantly at 99% confidence interval.

Fig. 9 E.coli cell number with reduction of water content in manure
fermented for 14 days

CONCLUSIONAs the loss of E.coli from farmlands is big concern from the viewpoints of environmental
conservation, this study dealt with 2 kinds of experiments, one is for observing E.coli loss using
slope model plots under artificial rainfall simulator and the other is for observing survival limit ofE.coli in the manure.
It was proven that the loss of E.coli was observed not only from the plot applied fresh cow
dung but also from the plot applied manure fermented for 14 days or 60 days. Also, E.coli loss
increased in proportion to the loss of organic matter in sediments. So, it was considered that
attentions should be paid more to soil erosion process, as it accelerates E.coli loss from uplandfields.
Also, it was concluded that E.coli vanished at the temperature higher than 60 degrees
Celsius.The reduction of water content was not enough to eliminate E.coli. So, it became clear that
controlling temperature of manure is important for vanishing E.coli existing in manures.
This work was supported by KAKENHI (C) of Grant-in-Aid for Scientific Research(21580301) of Japan Society for the Promotion of Science (JSPS).

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