Matching Livestock Systems with Available Resources |
Citation of this paper |
Fifteen Mong Cai gilts weighing 46±3.9 kg at service were used in a Randomized Complete Block Design (RCBD), with five replications of three treatments: TW0, 100% of supplementary protein supplied by soybean meal; TW50, 50% of supplementary protein supplied by soybean meal and 50% by a mixture of ensiled taro leaves and water spinach, and TW100, 100% of supplementary protein supplied by a mixture of ensiled taro leaves and water spinach. In the pregnancy period the feed was restricted to 1.5% of live weight. In the lactation period the gilts were fed increasing amounts of the same diet up to five days after farrowing, and from then onwards feeds were offered ad libitum.
Total dry matter intake decreased slightly with increased amounts of the mixture of taro leaf silage and water spinach. Live weights at farrowing and weaning declined as the amount of the mixture of taro leaf silage and water spinach increased. The feed conversion ratios for treatments TW0, TW50 and TW100 were 3.09, 3.96 and 5.02 kg feed/kg gain, respectively. Live weight loss and percentage live weight loss in lactation, and interval from weaning to oestrus were not affected by diet (P>0.05). The number of piglets born alive and at weaning did not differ among treatments (P>0.05). However, live weights of the litter at birth and weaning, and weights of individual piglets declined as the foliages replaced soybean meal. Mortality to weaning ranged from 10.2 (TW0) to 7.5% (TW50) and was not affected by the treatments.
It is concluded that reproduction in the Mong Cai breed, measured as numbers of piglets born alive and weaned, and the interval from weaning to estrus was satisfactory when taro leaf silage and water spinach replaced soybean meal. However, weights of piglets at weaning decreased, with a linear trend from 35.9 to 25.1 kg as the soybean was replaced by the forages.
Key words: Mong Cai gilts, piglet performance, reproduction, Taro leaf silage, water spinach
Pigs are widely kept throughout the country of Laos, with 64 percent of all households involved in pig production (Kaufmann et al 2003). The number of pigs kept by smallholders varies between an average of 1.4 and 3.7 animals per household, depending on the region (Kaufmann et al 2003). Pigs are normally raised in a free-range system, and supplemented by rice bran and other household waste products (http://clubs.uow.edu.au/clubs_websites/oxfam/Why%20Pigs.pdf). There are some large-scale pig enterprises close to Vientiane, but these use purchased concentrate feeds, which are not a viable option for small-scale farmers in the remote upland areas.
In view of the increasing costs of concentrate feeds, and especially protein concentrates such as soybean and fish meal, recent research in Vietnam, Cambodia and Laos has been directed to the use of leaves from crops such as cassava (Hang and Preston 2005; Chhay et al 2005), sweet potato (An 2004; Chittavong and Preston 2006), and mulberry (Phiny et al 2003), and water plants such as water spinach (Men 2000; Ly 2002) and duckweed (Rodriguez and Preston 1996; Hang 1998). Recent research has shown that fresh water spinach was more palatable than cassava leaves for growing pigs, as reflected in higher total DM intake, and the proportion of the diet (47%) provided by the leaves. Digestibility of dry matter, organic matter, N and crude fibre was higher in diets with water spinach than in those with cassava leaves (Chhay and Preston 2005).
Taro leaves (Colocasia esculenta (L.) Schott) are rich in vitamins and minerals. They are a good source of thiamin, riboflavin, iron, phosphorus and zinc, and a very good source of vitamin B6, vitamin C, niacin, potassium, copper and manganese (http://en.wikipedia.org/wiki/Taro). Leaves from Taro (Colocasia esculenta (L.) Schott and Alocacia macrorrhiza) and New Cocoyam (Xanthosoma sagittifolium) are traditionally used in pig diets by small-scale farmers in many tropical countries. A report from Colombia (Rodríguez et al 2006) showed that weight gains in young pigs fed a sugar cane juice diet were the same when the supplementary protein was from a 50:50 mixture of fresh leaves of New Cocoyam and soybean meal compared with soybean meal as the only protein source. Recent research from Vietnam (Duyet et al 2006) showed that there were no differences in live weight loss during lactation and days to first oestrus in Mong Cai sows when all the soybean meal was substituted by a mixture of forage protein sources (water spinach, cassava and sweet potato leaves). In contrast, Large White sows showed a significant deterioration in these traits when all the supplementary protein came from the leaves (Duyet et al 2005).
The present experiment aimed to evaluate the effect of protein-rich leaves as replacement for soybean meal on the performance of Mong Cai gilts during gestation and lactation.
The experiment was carried out from June 2006 to January 2007 at the Faculty of Agriculture, National University of Laos (NUOL), situated about 32 km from Vientiane city, Lao PDR. The mean daily temperature in the area at the time of the experiment was 27 oC (range 22-32 oC).
Fifteen Mong Cai gilts, with an average live weight at service of 46±3.9 kg, were used in the experiment, and were followed for one complete reproductive cycle. The gilts were imported from Vietnam.
There were three treatments:
TW0:100% of supplementary protein supplied by soybean meal (no green foliage supplied).
TW50:50% of supplementary protein supplied by soybean meal and 50% by a mixture of ensiled taro leaves and water spinach (equal parts of each foliage on DM basis).
TW100:100% of supplementary protein supplied by a mixture of ensiled taro leaves and water spinach (equal parts of each foliage on DM basis).
The experiment was done according to a Randomized Complete Block Design (RCBD) with five replications of each treatment. The gilts were from five litters, with three gilts from the same litter randomly allocated to each treatment. All the gilts were of similar initial body weight.
The pigs were housed in individual pens made from wood, in an open-sided building with a thatch roof of Imperata grass. The size of each pen was 2.0*1.5m.
The energy component of the diet was broken rice and cassava root silage. The composition of the vitamin-mineral premix is shown in Table 1. The data on chemical composition of ingredients and diets are in Tables 2 and 3.
Leaves of taro were purchased in Nonveng village, Hatchaifong district, Vientiane City, and made into silage, using molasses (4%, fresh basis). This was found to be the most suitable level of molasses from the results of a previous study (Chittavong, 2007). The silage was stored for 21 days in plastic bags to ensure complete fermentation. Some of the water spinach was grown in the Faculty farm; but most was purchased from farmers in the area around the Faculty of Agriculture, National University of Laos. Prior to feeding it was chopped into small pieces and mixed with taro leaf silage and the other ingredients of the diet (cassava root silage and broken rice). The mixed feeds were given to the pigs in 3 meals daily (7:30, 11:30 and 16:30h). Water was permanently supplied through drinking nipples.
The Mong Cai gilts were mated at third estrus by natural mating with the same Mong Cai boar. The feeding level was at 4% of live weight (DM basis) until pregnancy was confirmed, after which it was restricted to 1.5% of live weight. In the lactation period the gilts were fed increasing amounts up to five days after farrowing, and from then onwards feeds were offered ad libitum.
The gilts were weighed at intervals of 2 weeks during pregnancy. They were weighed after farrowing and then every 2 weeks until the end of lactation (42 days). Piglets were weighed at birth and then every 2 weeks to weaning. Feed intake and live weight gain were recorded during the gestation and lactation periods. Litter size at birth and at weaning, birth weight and weaning weight, mortality of the piglets at birth, live weight changes of the sows during lactation, and interval from weaning to oestrus were also calculated.
Samples of taro leaves were taken at the time of ensiling. Samples of water spinach, cassava root and of broken rice were taken for analysis when they were purchased.
Feed samples were analysed for dry matter by micro-wave radiation (Undersander et al., 1993). Nitrogen and crude fibre were determined according to AOAC (1990).
The data were analyzed using the General Linear Models procedure of ANOVA in
the MINITAB 13.31 program (2000). Sources of variation were blocks,
treatments and error.
The taro leaf silage and fresh water spinach had DM contents of 20.2 and 8.19 %, respectively, and crude protein (CP) contents of 19.0 and 18.8 % on a DM basis, respectively. The chemical composition of the diets and planned ration formulation are shown in Tables 1 and 2.
Feed and nutrient intakes are presented in Table 3 and 4. The proportion of the diet CP derived from the individual ingredients during pregnancy is shown in Figure 1. The protein was supplied almost entirely by soybean meal and the mixture of taro leaf silage and water spinach. Total DM intake decreased slightly with increased amount of the mixture of taro leaf silage and water spinach. In pregnancy, the daily DM intake was highest in diet TW0 (946 g/day), which was higher (P<0.05) than in the other two diets (910 and 794g/day for diet TW50 and TW100, respectively) (Figure 2). The CP intake was highest in diet TW0 (85 g/day) (P<0.05) (Figure 3). In the lactation period daily DM and CP intakes were not different among diets (P>0.05).
Table 1. Composition of the vitamin-mineral premix supplied
|
|
Item |
Amount (per kg) |
Vitamin premix |
|
Vitamin A, million IU |
10.0 |
Vitamin D3, million IU |
2.50 |
Vitamin E, IU |
5000 |
Vitamin K3,g |
1.60 |
Vitamin B1, g |
1.20 |
Vitanin B2, g |
3.20 |
Vitamin B6, g |
1.20 |
Niacin, g |
5.00 |
Pantothenic acid, g |
4.00 |
Folic acid, g |
5.00 |
Biotin, g |
0.12 |
Vitanmin C, g |
30.0 |
Additives and preservatives, g |
12.0 |
Other, kg |
1.00 |
Mineral premix |
|
Manganese, g |
5.40 |
Iron, g |
14.2 |
Copper, g |
1.00 |
Zinc, g |
2.90 |
Sodium, g |
3.90 |
Iodine |
19.0 |
Potassium, mg |
0.90 |
Cobalt, mg |
1.10 |
Other, g |
1.00 |
Table 2. Chemical composition of ingredients (% dry basis)
|
||||||
Ingredient |
DM |
CP |
CF |
Ash |
Oxalic acid, % |
HCN, mg/kg DM |
Water spinach |
8.19 |
18.8 |
16 |
15.1 |
|
|
Taro leaf silage |
20.2 |
19 |
13.2 |
11.6 |
0.30 |
|
Soybean meal |
87.8 |
41.8 |
8.02 |
7.4 |
|
|
Cassava root silage |
43.7 |
1.12 |
4.12 |
3.5 |
|
325 |
Broken rice |
86.9 |
5.74 |
2.78 |
0.81 |
|
|
Salt |
95.1 |
|
|
|
|
|
Premix |
98.2 |
|
|
|
|
|
Table 3. Formulation of diets, % of DM
|
|
||
Ingredient |
TW0 |
TW50 |
TW100 |
Broken rice |
37.0 |
33.9 |
18.5 |
Cassava root silage |
49.5 |
44.6 |
48.0 |
Taro leaf silage |
0.00 |
7.00 |
16.0 |
Water spinach |
0.00 |
7.00 |
16.0 |
Soybean meal |
12.0 |
6.00 |
0.00 |
Premix |
1.00 |
1.00 |
1.00 |
Salt |
0.50 |
0.50 |
0.50 |
Total |
100 |
100 |
100 |
% CP |
10.0 |
10.0 |
10.0 |
Table 4. Effect of replacing soybean meal by a mixture of taro leaf silage and water spinach on daily feed and nutrient intakes of Mong Cai gilts during pregnancy and lactation
|
|||||
Parameter |
Dietary treatment* |
SEM |
P-value |
||
TW0 |
TW50 |
TW100 |
|||
No. of gilts |
5 |
5 |
5 |
|
|
Pregnancy period |
|
|
|
|
|
DM, g/day |
946a |
910a |
794b |
34.5 |
0.022 |
CP, g/day |
84.8a |
82.0a |
72.2b |
3.05 |
0.031 |
CF, g/day |
39.6b |
52.0a |
58.0a |
1.82 |
0.000 |
CP, g/kg DM |
89.8 |
90.6 |
90.2 |
0.42 |
0.440 |
DM/kg LW |
14.6 |
15.2 |
14.8 |
2.80 |
0.340 |
Lactation period |
|
|
|
|
|
DM, g/day |
2549 |
2434 |
2111 |
261.4 |
0.492 |
CP, g/day |
244 |
236 |
203 |
25.4 |
0.493 |
CF, g/day |
142 |
171 |
182 |
17.2 |
0.277 |
CP, g/kg DM |
98.8 |
98.0 |
100 |
1.42 |
0.620 |
DM/kg LW |
38.0 |
37.4 |
29.8 |
4.08 |
0.340 |
a,b Mean values within rows with different superscript letters are significantly different (P<0.05); * See Table 3. |
|
Figure 1. Proportion of diet crude protein derived from individual ingredients* during pregnancy * WS= water spinach; TLS= taro leaf silage; SBM= soybean meal; BR= Broken rice and CRS= cassava root silage
|
|
Figure 3. Mean intakes of crude protein (g/kg DM) during pregnancy and lactation |
Live weight changes of the Mong Cai gilts are presented in Table 5 and Figure 4. There were no significant differences among treatments in live weight at service (P>0.05). However, live weights at farrowing declined as the proportion of the mixture of taro leaf silage and water spinach increased (P<0.05). The live weight increase of the gilts in treatment TW0 and average daily gain (ADG) throughout pregnancy were higher than in TW100 (P<0.01).
Table 5. Effect of replacing soybean meal by a mixture of taro leaf silage and water spinach on the performance of Mong Cai gilts in pregnancy |
|||||
Parameter |
TW0 |
TW50 |
TW100 |
SE |
P-value |
Live weight, kg |
|
|
|
|
|
At service |
47.1 |
46.8 |
44.7 |
1.71 |
0.579 |
After farrowing |
82.8a |
74.2ab |
63.1b |
3.84 |
0.012 |
Change in pregnancy |
35.7a |
27.4ab |
18.4b |
3.14 |
0.007 |
Pregnancy |
|
|
|
|
|
ADG, g |
313a |
242ab |
163b |
26.9 |
0.007 |
FCR, kg feed DM/kg gain |
3.09a |
3.96ab |
5.02b |
0.37 |
0.010 |
Length of pregnancy, days |
114 |
113 |
112 |
0.77 |
0.287 |
a, b Mean values within rows with different superscript letters are different at P<0.05 |
The length of pregnancy was 114.2, 113.4 and 112.4 days in treatments TW0, TW50 and TW100, respectively (P>0.05).
The mean feed conversion ratios (FCR) for treatments TW0, TW50 and TW100 were 3.09, 3.96 and 5.02 kg feed/kg gain, respectively (P<0.01).
Live weight at weaning of the sows was highest in diet TW0 (69.9 kg) and lowest in diet TW100 (56.5 kg) (P<0.05). Live weight loss and percentage of live weight loss in lactation were not affected by the diet (P>0.05) (Table 6). Live weight loss of the sows during lactation was positively and linearly related to the growth rates of their litters (Figure 4 and 5).
Table 6. Effect of replacing soy bean meal by a mixture of taro leaf silage and water spinach on weight changes in lactation |
|||||
|
TW0 |
TW50 |
TW100 |
SE |
P-value |
Live weight, kg |
|
|
|
|
|
After farrowing |
82.8a |
74.2ab |
63.1b |
3.33 |
0.009 |
At weaning |
69.9a |
68.0ab |
56.5b |
3.26 |
0.040 |
Weight loss in lactation period |
13.0 |
6.20 |
6.80 |
2.20 |
0.111 |
Weight loss/day |
0.31 |
0.15 |
0.16 |
|
|
% weight loss |
15.0 |
8.00 |
10.2 |
2.59 |
0.209 |
Weaning to oestrus, days |
6.60 |
6.60 |
6.40 |
0.24 |
0.804 |
a, b Mean values within rows with different superscript letters are different at P<0.05 |
|
Figure 4. Relationship between ADG of the litter and the weight loss of the sows in lactation |
|
Figure 5. Live weight changes of the sows throughout the reproductive cycle |
All experimental sows returned to oestrus within around 6 days after weaning, and mean weaning to estrus interval did not differ among treatment (P>0.05).
Litter size and piglet live weights at birth, 28 days and weaning are shown in Table 7. The total mean number of piglets born per litter was 10.8, 10.8 and 10.6 in treatments TW0, TW50 and TW100, respectively (P>0.05). The number of piglets born alive was not different (P>0.05) among treatments. Percentage mortality at birth was numerically higher on the TW100 diet but the differences were not significant (P= 0.55, 0.78 and 0.85 at birth, 28 days and at weaning). The total number of piglets, live weight and percentage mortality at 28 days were not different among treatments (P>0.05). The highest litter live weight was for the diet TW0 (21.4 kg/litter), and lowest in diet TW100 (13.1 kg/litter) (P<0.05).
The total mean number of piglets per litter at weaning was 9.20, 9.20 and 8.60 in TW0, TW50 and TW100, respectively (P>0.05). The total weight of piglets weaned was highest in diet TW0 (35.9 kg/litter) and lowest in diet TW100 (25.1 kg/litter) (P<0.05).
Table 7. Effect of replacing soybean meal by a mixture of taro leaf silage and water spinach in the gestation and lactation diet on piglet performance |
|||||
|
TW0 |
TW50 |
TW100 |
SE |
P-value |
At birth |
|
|
|
|
|
Total litter size |
10.8 |
10.8 |
10.6 |
1.09 |
0.989 |
Total litter size live born |
10.2 |
10.0 |
9.40 |
1.06 |
0.860 |
% mortality |
5.56 |
7.87 |
11.0 |
3.41 |
0.546 |
Total litter weight, kg |
5.12 |
5.96 |
4.95 |
0.54 |
0.405 |
Mean live weight, kg |
0.47 |
0.55 |
0.46 |
0.03 |
0.164 |
At 28 days |
|
|
|
|
|
Total litter size |
9.60 |
9.40 |
8.60 |
1.00 |
0.759 |
Total litter weight, kg |
26.5 |
22.9 |
18.1 |
2.56 |
0.105 |
% mortality |
5.72 |
5.48 |
8.44 |
3.26 |
0.780 |
Litter weight change, kg |
21.4 a |
16.9ab |
13.1b |
2.08 |
0.048 |
Mean piglet live weight, kg |
2.81 |
2.54 |
2.01 |
0.23 |
0.071 |
At weaning |
|
|
|
|
|
Total litter size at weaning |
9.20 |
9.20 |
8.60 |
0.98 |
0.883 |
Total litter weight at weaning, kg |
35.9a |
30.9ab |
25.1b |
1.80 |
0.009 |
Litter weight change, kg |
30.8a |
24.9ab |
20.1b |
1.56 |
0.004 |
Mean piglet live weight, kg |
3.96 |
3.58 |
2.85 |
0.33 |
0.110 |
% mortality, birth to weaning |
10.2 |
7.5 |
8.4 |
3.43 |
0.851 |
a, b Mean values within rows with different superscript letters are different at P<0.05 |
The use of a high level of inclusion of a mixture of taro leaf silage and water spinach in the diet of Mong Cai gilts could be expected to reduce the dry matter feed intake, mainly due to the bulkiness and possibly also due to lower palatability of this feed, as was the case. This is in agreement with Rodríguez et al (2006), who reported that the DM intake was reduced when the percentage of New Cocoyam in the diet was increased to 100%. Recent research from Tiep et al (2006) showed that the feed intake was highest for a diet with 10% Alocasia macrorrhiza leaves and lowest for a diet with 15% Alocasia macrorrhiza leaves, which suggested that the highest level of inclusion of ensiled Alocasia macrorrhiza leaves had a negative effect on palatability.
One sow in the treatment TW100 farrowed 5 days before the expected farrowing date, which resulted in a shorter mean length of pregnancy for TW100, although the difference compared to the other two treatments was not significant. The length of pregnancy in this experiment is similar to the results of Loc et al (2000), who also reported that the length of pregnancy was not different among treatments when increasing levels of ensiled forages were included in the diets of Mong Cai sows.
Weight losses of sows during the 42 days lactation were 0.31, 0.15 and 0.16 kg/day for TW0, TW50, and TW100, respectively, which is similar to the findings of Nga et al (2000), who reported that the mean weight loss of Mong Cai sows during a 49 day lactation was 12.6 kg (0.21 kg/day). The sows had been fed during pregnancy a diet with 10.3 % CP in DM, mainly derived from cassava leaf meal and water spinach. During lactation, voluntary feed intake of highly prolific sows is frequently inadequate to meet nutrient requirements for maintenance and milk yield, and these sows must mobilize fat and protein reserves (O’Grady et al 1985; Noblet et al 1990). Revell et al (1998) reported that body weight loss from sows during lactation depended on the protein content of the diet offered during lactation but not on body composition at farrowing. Sows offered a high protein diet during lactation (19.0% CP and 15.6 MJ DE/kg as fed) lost on average 4.3 kg during the four weeks of lactation (0.15 kg live weight/day), while those offered a low protein diet (7.9% CP and 15.5 MJ DE/kg as fed) lost on average 30.8 kg during lactation (1.1 kg/day). Low protein intake during lactation was also demonstrated to increase body protein loss (Jones and Stahly 1999) and to reduce reproductive performance (King and Williams 1984; Brendemuhl et al 1987; Yang et al 2000), but this was not the case in the present study, probably because of the lower nutrient requirements of indigenous sows such as the Mong Cai.
The estimated requirements for crude protein are presented in Table 8 for diets with an “ideal” protein (Speer 1990) compared with conventional diets based on maize and soybean (NRC 1988).
Table 8. Estimates of crude protein required for diets with an “ideal” protein (from Speer 1990) and conventional diets based on maize and soybean meal (from NRC 1988) for the Mong Cai gilts in the present study |
||||||||
|
Gestation |
Lactation |
||||||
LW, kg |
Crude protein, g/day |
LW, kg |
Crude protein, g/day |
|||||
Speer |
NRC |
This study |
Speer |
NRC |
This study |
|||
TW0 |
66.3 |
51 |
80.3 |
85.0 (62) |
74.8 |
160 |
214 |
244 (178) |
TW50 |
64.3 |
49.5 |
77.8 |
82.0 (66) |
68.5 |
147 |
196 |
236 (190) |
TW100 |
56.2 |
43.3 |
68 |
72 |
58 |
124 |
166 |
203 |
Figures in brackets are adjusted for the observed apparent crude protein digestibility |
In all the diets, the amounts of CP consumed were slightly above the recommended levels according to NRC (1988) and much higher than requirements for a diet with an “ideal” protein. However, if the reduced protein digestibility is taken into account the intakes in the present study would be midway between the requirements according to the ”ideal” protein and those of NRC (1988).
Although digestible energy (DE) contents of the diets were not calculated the low DE intakes both in pregnancy and lactation would probably also have contributed to the significantly lower LW gains in pregnancy and higher lactation losses of the sows on the TW100 treatment.
All sows returned to oestrus within 6 days after weaning, despite the low protein content of the diets, which is in agreement with the results of Mejia-Guadarrama et al (2002), who found that crossbred sows (Pietrain x (Large White x Landrace)) returned to oestrus within 9 days after weaning, and that the interval weaning to oestrus did not differ between experimental groups fed high or low levels of protein (20% CP and 1.08% lysine or 10% CP and 0.5% lysine). The results from this experiment contrast with those of Nga et al (2000) who reported that Mong Cai sows returned to oestrus within 15.8 days after weaning at 49 days, when fed diets with 10.3 % of CP from cassava leaf meal and water spinach during pregnancy.
Duyet et al (2006) reported that litter weight at 21 days was reduced from
28.3 to 23.7 in purebred piglets from Mong Cai sows when the soybean
meal was replaced by a mixture of fresh leaves (from cassava, water spinach
and sweet potato) but litter size was not affected. These finding are
similar to the results in the present study, where at 28 days the litter
weight was reduced from 26.5 to 18.1 kg. The greater difference in our study
may have been because creep feed was not offered until 30 days after birth.
Size of litter was not affected by the replacement of soybean with the
forages, which is in agreement with the findings of Duyet et al (2006).
However, in Baxuyen sows fed ensiled cassava roots as the energy source,
supplying fresh duckweed (11% in the DM of the diet) as partial replacement
of the protein supplement led to higher number of piglets born alive and
heavier litter weights at birth and at weaning (Men et al 1997). The results
imply that duckweed protein, as well as being highly digestible (Rodríguez
et al 1996), has a good array of essential amino acids. Also duckweed is a
rich source of vitamin A precursors, particularly carotene (Leng et al
1995), and vitamin A is vitally important for reproduction. It is
probable that in our study the vitamin A content in the control diet was
adequate to meet the requirement of the pregnant sows, and so there was no
improvement found in litter size when additional vitamin A was supplied as
green forage.
A mixture of taro leaf silage and water spinach can replace 100 % of soybean meal in pregnancy and lactation diets for Mong Cai gilts without affecting sow reproduction, measured as numbers of live piglets born and weaned, and the interval from weaning to oestrus. However, weights of piglets at weaning decreased with a linear trend as the soybean was replaced by the forages.
We are very grateful to the Swedish International Development Cooperation Agency, Department for Research Cooperation (Sida-SAREC) through the regional MEKARN Project, for the financial support of this study.
I would also like to thank the Faculty of Agriculture, National University of Laos for allowing and helping me to carry out this experiment. The authors thank Mr Bounlerth Syvilai for analytical assistance in the laboratory of the Faculty of Agriculture.
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