Livestock Research for Rural

Volume 10, Number 3, 1998

Development

Biodigester effluent versus manure, from pigs

or cattle, as fertilizer for duckweed (Lemna
spp.)
Le Ha Chau
Institute of Agricultural Sciences, Ho Chi Minh City, Vietnam

Abstract

The research had the following objectives: to document the observed improvements in nutritive
value (protein content) and yield of duckweed when biodigester effluent, rather than original
manure, is used to fertilize duckweed ponds; to determine optimum fertilizing level of manure
and effluent for duckweed; to determine if there are differences between manure and effluent
from cows versus pigs as fertilizer for duckweed ponds.

The treatments were:

 Source of manure / effluent: From cows (C) or pigs (P)

 Processing: Fresh manure (M) or effluent (F)
 Concentration of nitrogen in the medium (10, 20 and 30 mg N/litre)

The design was a factorial arrangement (2*2*3) and there were 4 replications in a completely
randomized layout. The experiment was conducted for 24 days in the rainy season (August
1998).  Plastic baskets (n=48) lined with plastic film were used as the experimental ponds. The
surface of the water in each basket had an area of 0.145 m² and with 10 cm depth the volume was
14.5 litres. Each container was inoculated with 40g (250g/m2) of duckweed (Lemna spp.). The
yield of duckweed was calculated by subtracting the inoculum from the total biomass production
measured every 24 hours and was expressed as fresh duckweed (DW) yield g/m2/d. Manure and
effluent were taken from two plug-flow tubular plastic biodigesters: one charged with cow
manure the other with pig manure. Manure and effluent were added daily in quantities calculated
to maintain pond nitrogen levels at approximately10, 20 and 30 mg/litre.

The results of the experiment showed that  with the same input of nitrogen, plants nutrients
derived from biodigester effluent supported higher concentrations of crude protein in duckweed,
than nutrients from raw manure. Manure and effluent from pigs tended to support higher
concentrations of crude protein in duckweed than when cows were the source of these inputs.
The optimum level of nitrogen in the pond water was in the range of 20 to 30 mg/litre. Root 
length of duckweed was inversely related with protein content.  Higher pH of the pond water in
the range pH 6.4 to 7.2 was associated with duckweed of higher protein content.
Key word: Biodigesters, effluent, manure, duckweed, biomass, nitrogen, protein

Introduction

About 80% of the Vietnam population is engaged in the farm sector. They are trying very hard
every day to maintain the productivity of their farm plots which are getting smaller every year.
Of course their initiatives are limited by poverty. It seems that farmers have themselves
considered everything in a good order that fits their needs for income, marketability of the
product, the climate and the available feed resources.

Social and natural changes require that people adapt to new activities. It is clear that while basing
these activities on the traditional practices, it is necessary to have improvements, eliminate
outdated practices and establish an improved system so that a better harmony can be reached. For
the farmers, the most suitable production system would appear to be that which continues to
satisfy the increasing needs of food for their own needs, sending the surplus to the market to buy
other commodities required by their families.

There are many animal feed resources in the tropics such as sugar cane, cassava roots and
foliage, leaves from leguminous and other multi-purpose trees, and water plants. With
appropriate balancing of the locally available feed resources it is argued that it is possible to have
higher levels of production per unit area compared with when only the conventional feeds are
used (Preston 1998). To achieve this goal, of optimizing the use of available of feed resources in
a locality to secure maximum income, the chosen feeding systems do not aim at achieving
maximum daily weight gain, but more importance is given to local availability and price, and to
efficiency of overall resource utilization.

Duckweed is one of the outstanding local resources in the tropics. It is probably the fastest
growing of all multi-cellular plants. It grows naturally on waste water and can double its weight
in 24 hours. It is unique amongst plants in that its protein content can be manipulated according
to the nitrogen content of the water in which it is growing (Leng et al 1995; Rodriguez and
Preston 1996a). This is important because it facilitates integration of duckweed ponds with
biodigesters. It is the ideal water plant to introduce into an integrated farming system because it
can use the nitrogen in the effluent coming from the biodigester to enrich its protein content to a
level only slightly lower than soya bean, approaching 40% (in dry matter).

The fact that protein yields of duckweed can be as high as 10 tonnes/ha/year (Preston 1998),
compared with less than one tonne per year for soya bean protein, highlights the potential value
of this plant at the farm level. The critical factor appears to be the protein content, which in turn
depends on the nutrient status of the medium on which it is grown. This relationship was shown
by Leng et al (1995) when they analyzed duckweed (Spirodela spp) grown on sewage water. The
protein content rose from 20% to almost 40% in dry matter as the N content of the water was
increased from 5 to 40 mg/litre.
Duckweed has been known for a long time as a potential food for humans and animals, and as a
source of natural products. Duckweed has been used as the only source of supplementary protein
for fish (PRISM 1997), chickens (Haustein et al 1990), ducks (Bui Xuan Men et al 1995) and
pigs (Rodriguez and Preston 1996b).

In Vietnam, the most common farm manures are from cattle, buffaloes and pigs. Fresh or
anaerobically fermented farm manures are rich sources of nutrients which can be used for
growing duckweed economically. Duckweed is good for the environment because it doesn't
require artificial fertilizers; on the contrary it cleans up waste by removing organic and inorganic
nitrogen coming from decomposition of organic matter, contributing to the fight against
eutrophication. It doesn't need fungicides and has no significant natural pests.

The development of a manure-based duckweed production system and its utilization as livestock
feed is essential for sustainable livestock farming in this country. Therefore, the present research
programme has been undertaken with the following objectives.

Objective

 To document the observed improvements in nutritive value (protein content) and yield of
duckweed when biodigester effluent, rather than original manure, is used to fertilize
duckweed ponds
 To determine optimum fertilizing level of manure and effluent for duckweed
 To determine if there are differences between manure and effluent from cows versus pigs
as fertilizer for duckweed ponds

Materials and methods

Location

The experiment was done at the "Finca Ecologica" on the Campus of the College of Agriculture
and Forestry, Thu Duc district, Ho Chi Minh city. This is a small (3,000 m²) experimental farm,
established to demonstrate integrated farming systems with perennial crops, multi-purpose trees,
local breeds of livestock, low-cost plastic biodigesters and duckweed ponds
(www.hcm.fpt.vn/inet/~ecofarm). The area is close to sea level with ranges in temperature from
24 to 38 ºC, and relative humidity in the range 40 to 100%.

Treatments and design

The treatments were:

 Source of manure / effluent: From cows (C) or pigs (P)

 Processing: Fresh manure (M) or effluent (F)
 Concentration of nitrogen in the medium (10, 20 and 30 mg N/litre)

The treatments were arranged as a 2*2*3 factorial and there were 4 replications in a completely
randomized block design. The experiment was conducted for 24 days in the rainy season (August
1998) The data were analyzed using analysis of variance (GLM) in Minitab Version Release
10.2. The sources of variation in the ANOVA were: animal species (cow vs pig), processing
(manure vs effluent), interaction (species*processing), pond N levels and error.

Management and data collection

Plastic baskets (n=48) lined with plastic film were used as the experimental ponds. The surface
of the water in each basket had an area of 0.145 m² and with 10 cm depth the volume was 14.5
litres. Each container was inoculated with 40g (250g/m2) of duckweed (Lemna spp.). The yield
of duckweed was calculated by subtracting the inoculum from the total biomass production
measured every 24 hours and was expressed as fresh duckweed ( DW) yield g/m2/d.

Manure and effluent were taken from two plug-flow tubular plastic biodigesters: one charged
with cow manure the other with pig manure. An explanation and description of the biodigester
system can be found in the paper by Bui Xuan An et al (1997)
(http://www.cipav.org.co/lrrd/lrrdhome.html). Fresh samples of manure and effluent were
sampled for DM and nitrogen. The amounts of manure and of effluent to be added to the ponds
were determined by the content of N found in these materials.

Samples of water and duckweed were collected at each harvest for determination of nitrogen and
dry matter. Root length of the duckweed was measured by extending 10 individual plants from
each sample on millimetric paper and taking the average length. Dry matter was determined by
weighing before and after drying to constant weight in a micro-wave oven (Undersander et al
1993) and nitrogen by Kjeldahl (AOAC 1985) using a Tekator apparatus. The samples of
duckweed were analyzed immediately after harvest because when the samples were stored in a
refrigerator there were obvious physical changes in their appearance and it was considered this
might affect the results.

Results and discussion

Composition of the different types of manure used on duckweed

Table 1 shows that dry matter and total nitrogen contents of manure and biodigester effluent
from pigs were higher than from cows.

Table 1. Dry matter (DM) and Nitrogen (N) content of different

types of manure were used on Duckweed

Type of manure Dry matter Total Nitrogen (% of DM)

(%)

Fresh cow dung (CM) 18.80 1.83

Fresh pig dung (PM) 24.05 2.40
Digested cow dung 3.12 1.65
(CF)
Digested pig dung 2.30 2.64
(PF)
 Nitrogen levels in pond water

Figure 1: Relationship between intended and Figure 2: Relationship between N in pond water
actual levels of nitrogen in pond water and biomass yield

On the whole, the intended levels of nitrogen in the pond water were achieved as demonstrated
by the mean values for this parameter during the trial (Figure 1). There was, however, a poor
relationship between measured levels of N in pond water and biomass yield (Figure 2). 

Effect of manure or effluent biomass yield of duckweed

It can be seen from Figure 3 that there was an interaction (P=0.001) between source of nutrients
(manure or effluent) and length of growing period. Manure supported higher biomass yield in the
first week but the rate of decline in yield with time was faster for manure than for effluent, thus
in the third week yields were higher for the effluent. These trends were the same for manure and
effluent from both cows and pigs. Increasing the application rate of the manure  / effluent (which
aimed to establish levels of 10, 20 and 30 mg N/litre) led to increased biomass yield with little
difference between the two higher levels (Figure 4). Overall, the manure supported slightly, but
significantly, higher yields than the effluent.
 Figure 4: Biomass yield of fresh duckweed fertilized
Figure 3: Biomass yield of fresh duckweed fertilized with biodigester effluent or manure from cattle or pigs
with biodigester effluent or manure from cattle or pigs at three intended levels of N in the water
averaged over three consecutive 7-day periods

Protein content of duckweed

Crude protein content of duckweed was higher (P=0.001) and root length shorter (P=0.001)
when the ponds were fertilized with effluent rather than manure (Figures 5 and 6). There were
increases (P=0.04) in crude protein content when the manure or effluent was from pigs rather
than cattle.  There was also a tendency (P=0.19) for root length to be shorter  with  manure and
effluent from pigs rather than cows. Root length was negatively correlated with protein content
of duckweed (Figure 7; R²=0.84) and with biomass yield (Figure 8; R²=0.40).

Figure 5: Effect of biodigester effluent or Figure 6: Effect of biodigester effluent or

manure from
manure from
cows and pigs on crude protein content of
cows and pigs on root length of duckweed
duckweed

Figure 7: Relationship between root length and Figure 8: Relationship between root length and
crude biomass
protein content of duckweed yield of duckweed

Discussion

The experimental procedure,  of varying input levels of manure and effluent so as to achieve
nitrogen concentrations in pond water in the range from 10 to 30 mg/litre, was largely successful.
However yield was poorly related with the nitrogen content of the pond water. This is in marked
contrast with results reported by Leng et al (1995), Rodriguez and Preston (1996a) and Nguyen
Duc Anh et al (1997) where the two elements were strongly and positively related.

The increase in protein content of duckweed when it was fertilized with biodigester effluent
compared with manure, from which the effluent was derived, is similar to what was observed in
cassava foliage from plants receiving similar treatments (Effluent or manure) (Le Ha Chau
1998).  This effect implies that during the process of anaerobic biodigestion the nitrogenous
compounds in the manure become more available to the plant (presumably the conversion from
organically-bound nitrogen to NH4+), and that the effect is similar in duckweed and in cassava. 
By contrast, the use of effluent as opposed to manure did not increase biomass yield of
duckweed but increased foliage biomass yield in cassava (Le Ha Chau 1998). There is no
apparent explanation for these contrasting effects. The indications of a relationship between pH
of the pond water and the crude protein content of the duckweed (r = +0.39)  and pH and the root
length (r = -0.32) could be interpreted as an effect of increased concentration of  NH4+ ions being
the determinant of the protein status of the duckweed.
The close relationship (R²=0.84) between root length of the duckweed and the protein content of
the whole plant is in agreement with earlier reports by  Rodriguez and Preston (1996) and
Nguyen Duc Anh et al (1997) and confirms the usefulness of this simple measurement to assess
the protein status of duckweed.  The tendency (R²=0.40) for biomass yield to be negatively
related with root length suggests that this latter parameter can also be used as an indicator of
yield, albeit an approximate one,  as well as quality.

Figure 9: Relationship between pH of pond Figure 10: Relationship between pH of pond

water and water and
crude protein in duckweed root length of duckweed

Conclusions and recommendations

The results of the experiment strongly indicate that:

 With the same input of nitrogen, plant nutrients derived from biodigester effluent support
higher concentrations of crude protein in duckweed, than nutrients from raw manure.
 Manure and effluent from pigs tended to support higher concentrations of crude protein
in duckweed than when cows were the source of these inputs.
 The optimum level of nitrogen in the pond water is probably in the range of 20 to 30
mg/litre.
 Root  length of duckweed is inversely related with protein content and is therefore an
excellent indicator of the protein status of the plant.
 Higher pH of the pond water in the range pH 6.4 to 7.2 is associated with duckweed of
higher protein content.
Acknowledgments

I gratefully extend my sincere thanks to Dr. Thomas R Preston for his valuable help in this study
and his dedication to the sustainable agriculture in developing countries. I also thank Lylian
Rodriguez for support in applying the technology and advice on practical problems of using the
biodigester. My thanks to Nguyen van Lai for advice on analytical procedures and assistance in
the laboratory. I also wish to express my gratitude to Dr. La Van Kinh and especially the staff of
the Department of Animal nutrition of Institute of Agricultural Science of South Vietnam for
creating a pleasant environment and research facilities. The Danish Embassy in Hanoi is
acknowledged for the financial support to the UTA foundation which made it possible for me to
carry out this research as partial fulfillment of the requirements for the Master of Science degree
in Sustainable Use of Local Resources in Livestock-based Farming Systems.

References

AOAC 1985 Official methods of analysis. Association of Official Agricultural Chemists. 12th
edition, Washington DC

Bui Xuan An, Preston T R and Dolberg F 1997 The introduction of low-cost polyethylene
tube biodigesters on small scale farms in Vietnam. Livestock Research for Rural Development
(9) 2:27-35(http://www.cipav.org.co/lrrd/lrrdhome.html)

Bui Xuan Men, Ogle B and Preston 1995 Use of duckweed (lemna spp) as replacement for
soya bean meal in a basal diet of brocken rice for fattening ducks. Livestock Research for Rural
Development. Volume 8, number 2: 5-8(http://www.cipav.org.co/lrrd/lrrdhome.html)

Haustein A T, Gilman R H, Skillicorn P W, Vergara V, Guevara V and Gastanaduy A 1990

Duckweed. A useful strategy for feeding chickens: performance of layers fed with sewage-grown
Lemnacea species. Poultry Science. (69):1835-1844

Le Ha Chau 1998 Biodigester effluent versus manure, from pigs or cattle, as fertilizer for
production of cassava foliage (Cassava Esculenta). Livestock Research for Rural Development.
Volume 10, number 3: submitted

Leng R A, Stamboli J H and Bell R 1995 Duckweed - A potential high protein feed resource
for domestic animals and fish. Livestock Research for Rural Development (7)
1:36kb(http://www.cipav.org.co/lrrd/lrrdhome.html)

Nguyen Duc Anh, Huu Tao and Preston T R 1997 Effect of management practices and
fertilization with biodigester effluent on biomass yield and composition of duckweed. Livestock
Research for Rural Development (9) 1:46-51(http://www.cipav.org.co/lrrd/lrrdhome.html)

Preston T R 1998 Animal production and use of natural resources. Proceedings 8th World
Conference on Animal Production.(In press):
PRISM 1997 Economic feasibility of duckweed-based fish production: a few case studies.
Duckweed Research Project. Report 1097, June 1997

Rodríguez Lylian and Preston T R 1996a Use of effluent from low cost plastic biodigesters as
fertilizer for duckweed ponds. Livestock Research for Rural Development (8) 2:60-
69(http://www.cipav.org.co/lrrd/lrrdhome.html)

Rodriguez Lylian and Preston T R 1996b Comparative parameters of digestion and N

metabolism in Mong Cai and Mong Cai Large White cross piglets having free access to sugar
cane juice and duckweed Livestock Research for Rural Development (8) 1:72-81
(http://www.cipav.org.co/lrrd/lrrdhome.html)

Undersander D, Mertens D R and Thiex N 1993 Forage analysis procedures. National Forage
Testing Association. Omaha. pp: 154

Received 11 December 1998

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