Citation |
Two experiments were conducted at CelAgrid, Kandal Province for a period of 80days. In experiment 1, 12 ponds each with area of 10m2 were used to compare four treatments arranged in a 2 x 2 factorial with a completely randomized design with 3 replicates. The first factor was fertilizer with effluent and no effluent; the second factor was stocking density of 3 or 5 fish per m2. The fish in each pond were provided with feed at 5% of the fish biomass (DM basis). The feed contained 25% of water spinach, 25% of duck weed and 50% of rice bran (DM basis). The effluent from a bio-digester was applied every 7 days, at rates equivalent to 150 kg N/ha.
Survival rate was higher in ponds fertilized with effluent and in ponds with lower fish density. There were no differences for gain in weight and length due to treatment, and no interaction between fertilizer and stocking density. However, these data were derived from random samples of fish taken at 20 day intervals and the results were partially confounded by differences among treatments in survival. Net fish yield was higher for the higher stocking density but there was no effect of fertilization with effluent. Feed conversion (DM offered/net fish yield) did not differ among treatments but this measurement was also confounded by differences in survival as amounts of feed offered were based on initial numbers of fish and the average weight estimated from the sampling at 20 day intervals. Final fish weight and net fish yield were negatively correlated with survival rate.
In experiment 2, 12 plots in a paddy field each with an area of 209m2 were used to compare 4 treatments in a 2 x 2 factorial in a completely randomized design with 3 replicates. The first factor was with or without feed supplement; the second factor was different stocking densities of 3 or 5 fish per m2. In each plot of paddy there was a trench 11m wide x 1m in length x 1m deep along one side of each plot. The feed supplement was the same as in experiment 1. All paddy plots were fertilized with effluent from a bio-digester every 7 days at the rate of 150 kg N per ha.
Survival rate was not affected by supplementation but there was a tendency (P = 0.10) for it to be lower on the higher stocking rate. Both final weight of fish and the net fish yield were increased by supplementation and by stocking rate with no interaction between the treatments. The FCR (for those paddies that received feed supplementation) was not affected by stocking rate.
Cambodian people prefer their protein to come from fresh water fish that is eaten fresh, salted, smoked or made into fish sauce and paste. Tonle Sap, Tonle Mekong and Basak rivers are the main capture fisheries in Cambodia (Agriculture in Cambodia 2010).
Tilapia has become popular for farmers as it is easy to culture and there is a good demand in the market. Moreover, tilapias (Oreochromis spp.) adapt well to the local environment and local feed, and have high productivity. The fish are usually kept in a pond near to their houses, as in addition to having the fish as protein source, farmers can grow vegetables and use the water from the pond to water the vegetables. The feeds used for the fish depend on the resources available in the area. Duckweed and water spinach are available almost everywhere in the villages; while rice bran is the byproduct from the rice milling.
The combination of rice and fish can be a very profitable system, since it was observed that the fish feed on organisms, such as insects and larvae which grow and live in the rice fields. This system provides both rice and fish. Besides economic benefits, the biological benefit is also a factor. Weeding and use of chemical fertilizers and pesticides are reduced when this system is practiced. Moreover, the movement of the fish stirs the water, which increases the oxygen level and improves the development of the roots of the rice. Rice-fish culture improves the income of farmers in the rural areas, as the system requires very little inputs and farm labor. Farmers can harvest rice or fish at the same time or harvest only the rice and keep the fish, or alternatively harvest the fish before the rice (Mackay 1995).
In pond culture the net fish yield of tilapia will be increased by fertilizing with bio-digester effluent and by increasing the stocking density.
When tilapia is raised in the rice paddy there will be advantages from supplementation with duckweed, water spinach and rice bran.
Two experiments were carried out to study the effects of stocking density and fertilization/supplementation on the growth performance of Tilapia raised in ponds and in a paddy field.
The experiments were carried out at the Center for Livestock and Agriculture Development (CelAgrid) experimental farm, located in Prah Theat village, Rolous commune, Kandal Steung district, Kandal Province, approximately 19 km from Phnom Penh City.
In Cambodia, the rainy season is from June to October, while the dry season is from November to May. This climate provides good conditions for the animal and rice production system. Average temperature is around 25oC, with a maximum of about 40oC in April, while the coldest month is January, when the temperature is around 21oC, with a maximum of about 31oC. Average annual rainfall in Cambodia varies from 1,500 mm or less in the central plain and 1,500 to 2,500 mm in the surrounding mountains. Over most of the South West coastal region, average annual rainfall is in excess of 3,000 mm. The rainfall in the East of the Mekong River is generally between 1,800 mm and 3,000 mm, while the lower Mekong valley and basin of the Tonle Sap Lake is relatively dry, with rainfall averaging between 1,200 and 1,500 mm.
Materials and methods
The experiment was conducted from 12 January to 1 April 2010.
A total of 12 ponds were prepared, each with an area of 10 m2 (4 m in length x 2.5 m wide) and a depth of 1.5 m at the CelAgrid centre. The ponds were lined with plastic to avoid filtration of water, and then water was pumped in from nearby canals and pond. Lime (CaO) at 200 g/m˛ was applied before stocking the fish, in order to kill parasites and pathogenic organisms and also to increase water pH.
The experiment was conducted 80days and designed as 2 x 2 factorial arrangements: the factors were with or without application of biodigester effluent, and two stocking densities (3 and 5 fish per m2) (Table 1). A Complete Randomized Design (CRD) was used. Each treatment was replicated 3 times. In total 640 fingerlings were bought from a commercial fish farm and randomly distributed into the ponds. Weight and length of a sample of the fish were recorded as the initial weight and length.
Table 1: Experimental layout |
|||||||||||
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
5F-NE |
3F-E |
5F-E |
3F-E |
3F-NE |
5F- NE |
3F-NE |
3F-E |
5F-NE |
5F-E |
5F-E |
3F-NE |
F = Feed, E = Effluent, NE = Not fertilized with effluent; 3 and 5 = density 3 or 5 fish/m2 |
The fish were fed a mixture of rice bran, water spinach and duckweed as the basal feed. All ponds were provided with the same feed (Table 2).
Table2: Ingredient composition of the diet, % DM basis |
|
Water spinach |
25 |
Duckweed |
25 |
Rice bran |
50 |
Total |
100 |
% Crude protein (in DM) # |
18.5 |
# Calculated on basis of observed composition of the ingredients (Table 3) |
Duckweed was cultivated in CelAgrid, while water spinach was bought from a local market. In order to reduce the moisture content, duckweed was collected in the evening and wilted for the morning feeding, while the duckweed collected in the morning was wilted and fed to the fish in the evening. Water spinach was chopped into small pieces and mixed with duckweed and rice bran before feeding. The feed was provided twice daily at 8:00h and 16:00h. The amount of feed was 5% (DM basis) of the fish body weight. The amount of feed was adjusted by the average of fish sampling multiply with the initial number of the fish at the beginning. The feed was mixed and put in a floating feeding frame to avoid the feed spreading in the pond (Photo 1).
Ponds were fertilized with effluent from a plastic bio-digester loaded with pig manure, at a rate of 150 kg of N/ha/year (San Thy et al 2006). The effluent from the bio-digester was pumped into containers (Photo 2). Before applying it to the fish pond, a sample was taken to determine N. The amount of effluent applied to the ponds was calculated according to the concentration of N.
Photo 1: Individual pond with floating feeding frame |
Photo 2: Effluent storage |
The oxygen and pH of the water in the fish pond were measured every 5 days. Each measurement was taken 2 times at 06:00h and 16:00h using a pH meter (pHep by HANNA) and a DO2 meter (Model 9150), respectively. The water temperature was measured 3 times every 5 days at 06:00h, at 12:00h and at 16:00h using thermometers. A thermometer was placed permanently in each pond. Water transparency was measured at 12.00h every 2 days using a Secchi disk.
Every 20 days a sample of the fish was caught with a seine net and ten individuals chosen at random. These were weighed using an electronic scale and measured with a ruler from the mouth tip to the caudal fin. Survival rate was measured at the end of the experiment by the following equation:
X (%) = (Nt / N0) x 100
Where: N0 : initial number of the fish; Nt : final number of the fish
The data were subjected to analysis of variance (ANOVA) by using the General Linear Model (GLM) of the Minitab software (version 2000 release 13.1). Sources of variation were: effluent, density, effluent * density interaction and error. Gain in weight and length were measured as the linear regression of weight (or length) on days in the experiment, using the SLOPE command in the Minitab software.
Table 3: Chemical composition of the diet ingredients |
||
|
DM, % |
CP, % of DM |
Water spinach |
13.6 |
26.0 |
Duckweed |
6.86 |
30.5 |
Rice bran |
90.2 |
8.65 |
Effluent from the biodigester was analyzed for N, and the amount calculated based on the rate of N application of 2.88 g N per pond per week (Table 4).
Table 4: Nitrogen requirement in each pond # |
|
Application level of N, kg/ha |
150 |
Area of pond, m2 |
10 |
N requirement, g/pond |
150 |
N requirement per pond per week, g |
2.88 |
# Source: San Thy et al 2006 |
The treatments had no effect on water quality, measured by pH, temperature or
dissolved oxygen levels (Table 5), all of which were within the normal range for
culture of Tilapia (Swingle 1969). There was an interaction between treatments
for water density (Table 6), which was more transparent (less phytoplankton)
with the lower fish stocking rate when effluent was applied. In contrast, in the
absence of effluent, the water was more transparent at the higher stocking rate.
.Table 5: Mean values for water quality in ponds stocked with Tilapia at different densities and fed supplements of rice bran, water spinach, and duckweed with addition of bio-digester effluent or none |
||||||||
|
Effluent (E) |
Density (D) |
Probability |
SEM |
||||
Effluent |
No effluent |
3 |
5 |
E |
D |
E*D |
||
pH |
||||||||
06:00h |
7.8 |
7.9 |
7.9 |
7.9 |
0.79 |
0.98 |
0.77 |
0.058 |
16:00h |
8.8 |
8.1 |
8.8 |
8.1 |
0.30 |
0.30 |
0.30 |
0.514 |
DO, mg/liter |
||||||||
06:00h |
3.1 |
3.0 |
3.0 |
3.1 |
0.74 |
0.63 |
0.75 |
0.104 |
16:00h |
4.2 |
4.2 |
4.1 |
4.3 |
0.77 |
0.28 |
0.73 |
0.155 |
Temperature, o C |
||||||||
06:00h |
28.9 |
28.8 |
28.8 |
28.8 |
0.49 |
0.94 |
0.31 |
0.106 |
12:00h |
31.5 |
31.4 |
31.4 |
31.4 |
0.68 |
0.85 |
0.12 |
0.199 |
16:00h |
33.1 |
33.0 |
33.1 |
33.0 |
0.72 |
0.68 |
0.38 |
0.201 |
Water transparency, cm |
||||||||
12:00h |
18.1 |
18.7 |
18.7 |
18.1 |
0.29 |
0.26 |
0.001 |
0.380 |
Table 6: Mean values for water density in ponds stocked with Tilapia at different densities and fed supplements of rice bran, water spinach, and duckweed with addition of bio-digester effluent or none |
||||||
Effluent |
Yes |
No |
SEM |
P |
||
Fish. m2 |
3 |
5 |
3 |
5 |
||
Water transparency, cm |
19.8a |
16.6b |
17.7b |
19.7a |
0.53 |
0.001 |
a, b Mean values without common superscript differ at P<0.05 |
Table 7: Mean values of live weight of tilapia fertilized with and without effluent |
||||||||
|
Effluent (E) |
Density (D) |
Probability |
SEM |
||||
|
Effluent |
No effluent |
3 |
5 |
E |
D |
E*D |
|
DWG, g/day |
0.700 |
0.607 |
0.698 |
0.608 |
0.223 |
0.238 |
0.891 |
0.049 |
DLG, mm/day |
0.102 |
0.092 |
0.097 |
0.097 |
0.320 |
1.000 |
0.733 |
0.006 |
W: L ratio |
0.044 |
0.039 |
0.041 |
0.042 |
0.211 |
0.793 |
0.544 |
0.002 |
Table 8: Mean values (main effects) for total weight gain, feeds offered and feed utilization for Tilapia (Oreochromis spp.) stocked at different densities and fed supplements of rice bran, water spinach, and duckweed, with addition of bio-digester effluent or none |
|||||||
|
Effluent (E) |
Density (D) |
Probability |
SEM |
|||
|
Effluent |
No effluent |
3 |
5 |
E |
D |
|
Water spinach, g/pond/d |
15.2 |
14.8 |
11.4 |
18.6 |
0.914 |
0.035 |
2.34 |
Duck weed, g/pond/d |
15.2 |
14.8 |
11.4 |
18.6 |
0.914 |
0.035 |
2.34 |
Rice bran, g/pond/d |
9.47 |
9.19 |
7.12 |
11.54 |
0.90 |
0.051 |
1.55 |
Total DM, g/pond/d |
39.8 |
38.8 |
29.9 |
48.7 |
0.91 |
0.038 |
6.23 |
Total DM, g/pond in 80d |
3183 |
3103 |
2390 |
3895 |
0.841 |
0.005 |
273 |
Initial wt, g/pond |
142 |
162 |
123 |
180 |
0.203 |
0.004 |
10.2 |
Final wt, g/pond |
2010 |
2006 |
1589 |
2427 |
0.986 |
0.006 |
161 |
Net fish yield, g in 80 days |
1869 |
1845 |
1466 |
2248 |
0.918 |
0.009 |
160 |
FCR |
1.71 |
1.71 |
1.67 |
1.74 |
0.999 |
0.725 |
0.131 |
Survival, % |
74 |
64 |
78 |
60 |
0.045 |
0.003 |
3.03 |
FCR = Feed DM offered/net fish yield |
Table 9: Mean values for weight gain, feeds offered and feed utilization for Tilapia (Oreochromis spp.) stocked at different densities and fed supplements of rice bran, water spinach, and duckweed with addition of bio-digester effluent or none |
||||||
|
Effluent |
No effluent |
SEM |
P |
||
|
3 |
5 |
3 |
5 |
||
Water spinach, g/pond/d |
11.6 |
18.7 |
11.1 |
18.5 |
3.31 |
0.208 |
Duck weed, g/pond/d |
11.6 |
18.7 |
11.1 |
18.5 |
3.31 |
0.208 |
Rice bran, g/pond/d |
7.28 |
11.67 |
6.96 |
11.4 |
2.20 |
0.27 |
Total DM, g/pond/d |
30.5 |
49.0 |
29.2 |
48.4 |
8.81 |
0.221 |
Total DM, g/pond in 80d |
2444 |
3922 |
2336 |
3869 |
386 |
0.94 |
Initial wt, g/pond |
116 |
167 |
130 |
193 |
14.43 |
0.656 |
Final wt, g/pond |
1672 |
2348 |
1505 |
2507 |
228.74 |
0.496 |
Net fish yield, g |
1556 |
2182 |
1375 |
2314 |
227.3 |
0.511 |
FCR |
1.61 |
1.81 |
1.74 |
1.68 |
0.19 |
0.888 |
Survival, % |
87.7 |
60.0 |
68.0 |
59.3 |
4.29 |
0.058 |
FCR = Feed DM offered/net fish yield |
Figure 1. Relationship between survival rate and final weight per fish |
Figure 2. Relationship between survival rate and net fish yield per pond |
Figure 3. Relationship between survival rate and DM offered per pond |
Figure 4. There was no relationship between survival rate and FCR (DM offered /net fish yield per pond) |
The net fish yield in this experiment (a range of 1500 to 2200 kg/ha) was twice as high as the yields (760 – 1200 kg/ha) reported by San Thy and Preston (2003). These researchers also used Tilapia but in ponds of 6m2 and at a lower stocking rate of 2 fish/m2. They applied biodigester effluent to the ponds at a similar rate (160 kg N/ha) but the fish received no supplementary feed. Nguyen Duy Quynh Tram et al (2007) fertilized ponds with raw pig manure or bio-digester effluent (derived from the same manure) at 240 kg N/ha over 120 days. The net fish yield of a mixture of Tilapia, Silver carp and Hybrid Catfish was 1700 kg/ha with the effluent and 2100 kg/ha with the raw manure.
· The net yield of fish was not affected by addition of effluent to the ponds but was higher for the greater stocking density.
· Survival rate was higher in ponds fertilized with effluent and in ponds with lower fish density.
· As the survival rate increased the final weight per fish and the net fish yield both decreased
The experiment was conducted in the dry season, from 23rd January to 13th April 2010.
Twelve plots were prepared in a paddy field. The total area of each plot was 11m x 19 m with a trench 11m wide x 1m in length x 1m deep at one side of each plot. In total 528 fingerlings were purchased from a fish hatchery farm, Preak Phnov, near Phnom Penh City. They were raised in a nursery pond in CelAgrid for 15 days before being introduced into the plots of paddy rice, which was done 7 days after rice transplanting.
|
||
Photo 3: Rice transplant |
Photo 4: Rice shooting |
Photo 5: Rice maturity |
The experiment was carried out as a 2 x 2 factorial arrangement. The factors were: with and without supplementary feed; and stocking densities of 3 and 5 fish per m2 (Table 10). A Complete Randomized Design (CRD) was used. Each treatment was replicated 3 times.
Table 10: Experimental layout |
|||||||||||
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
3E-F |
3E-NF |
3E-F |
5E-NF |
5E-F |
5E-NF |
5E-F |
3E-NF |
5E-NF |
3E-NF |
3E-F |
5E-F |
E = Effluent, F = Feed supplement, NF = No feed supplement, 3 and 5 = density 3 or 5 fish/m2 |
Fish were fed on a mixture of rice bran, water spinach and duckweed (Photo 6, 7 and 8; Table 11).
Table 11: Ingredient composition of the diet, % DM basis |
|
Water spinach |
25 |
Duckweed |
25 |
Rice bran |
50 |
Total |
100 |
% Crude protein (in DM)# |
18.5 |
# Calculated on basis of observed composition of the ingredients (Table 12) |
The feed was provided twice daily at 8:00h and 16:00h at an estimated rate of 5% (DM basis) of the fish body weight. The amount of feed was adjusted by the average of fish sampling multiply with the initial number of the fish at the beginning. Duckweed was cultivated in CelAgrid, while water spinach was bought from the market. In order to reduce the moisture content, duckweed was collected in the evening and wilted for the morning feeding, while duckweed collected in the morning was wilted and fed to the fish in the evening. Water spinach was chopped into small pieces and mixed with rice bran and duckweed before feeding to the fish, in the feeding frame (Photo 1). During successive periods of 20 days, water spinach and duckweed were harvested at the same place to make sure that the nutrient contained in water spinach and duckweed were not so much different.
Photo 5: Duckweed |
Photo 6: Chopped Water spinach |
Photo 7: Rice bran |
Fish weight and length were measured on random samples (n=10) of fish taken at 08:00h every 20 days before they were given feed and before application of effluent. Dry matter and CP of feeds were analyzed every 20 days before the fish sampling. Survival rate was measured at the end of the experiment. Water quality was measured following the same procedures as in Experiment 1. The fish were harvested after 80 days. Total weight, length and the number of the fish were measured.
The data were subjected to analysis of variance (ANOVA) by using the General Linear Model (GLM) of the Minitab software (version 2000 release 13.1). Sources of variation were: feed, density, feed * density interaction and error.
The crude protein content in duckweed was higher than in water spinach (Table 12).
Table 12: Chemical composition of the diet ingredients |
||
|
DM, % |
CP % of DM |
Water spinach |
13.6 |
26.0 |
Duckweed |
6.86 |
30.5 |
Rice bran |
90.2 |
8.65 |
All the ponds in the paddy field were fertilized with effluent from a bio-digester at a rate of 150 kg N per hectare/year (San Thy et al 2006). The amount of effluent was calculated on the basis of its content of N (Table 13). It was supplied directly into each paddy at intervals of 7 days.
Table 13: Total Nitrogen requirement in each pond # |
|
Proportion of N, kg/ ha |
150 |
Area of pond, m2 |
11 |
N requirement, g |
165 |
N requirement per week, g |
3.17 |
# Source: San Thy et al 2006 |
Table 14: Mean values for indices of water quality (main effects) |
||||||||
|
Supplement (S) |
Density (D) |
Probability |
SEM |
||||
|
Feed supplement |
No feed supplement |
3 |
5 |
S |
D |
S*D |
|
pH |
||||||||
06:00h |
8.0 |
8.0 |
8.0 |
8.0 |
0.540 |
0.858 |
0.664 |
0.057 |
16:00h |
8.2 |
8.3 |
8.3 |
8.2 |
0.691 |
0.794 |
0.937 |
0.064 |
DO, mg/liter |
||||||||
06:00h |
3.2 |
3.2 |
3.2 |
3.2 |
0.659 |
0.792 |
0.642 |
0.090 |
16:00h |
4.3 |
4.2 |
4.3 |
4.2 |
0.668 |
0.477 |
0.775 |
0.104 |
Temperature, o C |
||||||||
06:00h |
27.9 |
27.9 |
28.0 |
27.9 |
0.9955 |
0.691 |
0.709 |
0.116 |
12:00h |
30.4 |
30.5 |
30.5 |
30.4 |
0.642 |
0.636 |
0.877 |
0.241 |
16:00h |
32.0 |
32.0 |
32.1 |
31.9 |
0.801 |
0.681 |
0.534 |
0.195 |
Water transparency, cm |
||||||||
12:00h |
11.6 |
10.7 |
10.8 |
11.5 |
0.102 |
0.149 |
0.541 |
0.353 |
Survival rate was not affected by supplementation but there was a tendency (P = 0.10) for it to be lower on the higher stocking rate (Table 15). Both final weight of fish and the net fish yield were increased by supplementation and by stocking rate with no interaction between the treatments. The FCR (for those paddies that received feed supplementation) was not affected by stocking rate.
Table 15: Mean values for initial and final fish numbers, survival rate, feed offered, total weight gain and feed conversion for Tilapia in the paddy field at two densities and with and without supplements of duckweed, water spinach and rice bran |
|||||||
|
Feed supplement |
Prob. |
Fish/m2 |
SEM |
Prob. |
||
Supplement |
No supplement |
3 |
5 |
||||
Initial number |
44 |
44 |
|
33 |
55 |
|
|
Final number |
22.2 |
22.7 |
0.931 |
21.8 |
23.0 |
3.93 |
0.839 |
Survival, % |
51.2 |
56.8 |
0.686 |
66.2 |
41.8 |
9.4 |
0.104 |
Growth rate, g/day # |
0.677 |
0.458 |
0.010 |
0.573 |
0.562 |
0.046 |
0.863 |
Initial weight, g |
716 |
705 |
|
501 |
919 |
85.2 |
|
Final weight, g |
3256 |
2406 |
0.003 |
2174 |
3488 |
142.7 |
0.001 |
Net fish yield, g |
2540 |
1702 |
0.016 |
1672 |
2570 |
196 |
0.012 |
Net fish yield, kg/ha |
122 |
81.4 |
|
80 |
123 |
|
|
DM offered (80days) per paddy, g ## |
|
|
|
5802 |
9116 |
|
|
FCR, DM offered/net fish yield # |
|
|
|
2.85 |
3.02 |
|
|
# Based in weights of samples of fish taken at 20 day intervals ## Data are for the paddies that received the feed supplement |
Table 16: Reports from other research on rice-fish systems compared with data from the present study |
||||
Systems |
Treatments |
Growth rate, g/day |
Net fish production, kg/ha |
Authors |
Rice – fish (No feed supplement) with different seeding rate/ha |
100kg rice/ha |
0.80 |
32.5 |
Rothuis, 1998a |
200kg rice/ha |
0.89 |
23.8 |
||
300kg rice/ha |
0.80 |
16.7 |
||
Rice - fish Poly-culture with different stocking density (No feed supplement) |
6600fish/ha |
0.25 |
177.4 |
Rothuis 1998b |
5400fish/ha |
0.28 |
125.1 |
||
3400fish/ha |
0.33 |
110.5 |
||
1400fish/ha |
0.59 |
53.8 |
||
3800fish/ha |
0.25 |
62.4 |
||
Rice – fish (supplement rice bran) |
6000fish/ha |
0.48 |
132 |
Rasowo et al 2006 |
Rice – fish with different stocking density (rice bran 72%, copra meal 20% and soybean meal 8%) |
1500fish/ha |
|
508 |
Bocek |
3000fish/ha |
|
913 |
||
4500fish/ha |
|
1044 |
||
Rice-fish culture with different stocking density # |
Supplement |
0.67 |
122 |
This study |
No supplement |
0.45 |
81.4 |
||
# Mean values for densities of 1578 and 2631 fish/ha |
As the survival rate increased the final weight per fish and the net fish yield both decreased
The authors are grateful to the MEKARN program, financed by Sida (Swedish International Development Agency) for supporting this study.
Agriculture in Cambodia 2010: http://en.wikipedia.org/wiki/Agriculture_in_Cambodia#Rice_production
AOAC 1990: Official Methods of Analysis. Association of Official Analytical Chemists. 15th edition (K Helrick, editor). Arlington pp 1230